CN1867348B - Orally administered small peptides synergized statin activity - Google Patents

Abstract

This invention provides novel peptides that ameliorate one or more symptoms of atherosclerosis. The peptides are highly stable and readily administered via an oral route. The peptides are effective to stimulate the formation and cycling of pre-beta high density lipoprotein-like particles and/or to promote lipid transport and detoxification. This invention also provides a method of tracking a peptide in a mammal. In addition, the peptides inhibit osteoporosis. When administered with a statin, the peptides enhance the activity of the statin permitting the statin to be used at significantly lower dosages and/or cause the statins to be significantly more anti-inflammatory at any given dose.

Description

Orally administered small peptides synergize statin activity

The cross reference of related application

The application is the part continuity application of the USSN 10/649,378 of submission on August 26th, 2003, and USSN 10/649,378 has required the priority of the USSN 60/494,449 of submission on August 11st, 2003, and they are incorporated herein in full for various purposes.

Statement about the right of the invention under the research and development of federal government patronage, finished

This work is can authorize (National Heart by U.S. public health bureau (United States Public HealthService) and national heart, lung and hematology, Lung, and BloodInstitute Grants) HL30568 and HL34343 supported.U.S. government has some power in the present invention.

Invention field

The present invention relates to the atherosclerosis field.Especially, the present invention can be Orally administered and improve a class peptide of atherosclerotic one or more symptoms about identifying.

Background of invention

Cardiovascular disease is morbidity and main causes of death, particularly in the U.S. and the Western European countries.Relate to several causative factors in the development of cardiovascular disease, comprise heredodiathesis, sex, life style factor disease, for example smoking and meals, age, hypertension and hyperlipemia comprise hypercholesterolemia.Several in these factors, particularly hyperlipemia provides the significant risk factor relevant with atherosclerosis with hypercholesterolemia (high blood cholesterol levels concentration).

Cholesterol is present in the blood as the free and esterified cholesterol of lipoprotein particle inside, is commonly called Chylomicron, very low density lipoprotein (VLDL) (VLDL), low density lipoprotein, LDL (LDL) and high density lipoprotein (HDL).Total cholesterol concentration in blood is subjected to following influence: (1) is from gastral cholesterol absorption, (2) from the diet composition, for example carbohydrate, albumen, fat and alcoholic acid cholesterol are synthetic, (3) by tissue, particularly liver removes de-cholesterol and subsequently cholesterol is changed into bile acid, steroid hormone and biliary cholesterol from blood.

Blood cholesterol levels concentration keep the influence that is subjected to the h and E factor.Inherited genetic factors comprises the rate-limiting enzyme concentration in the cholesterol biosynthesis, and liver is in synthetic and the excretory speed and the people's of the concentration of low density lipoprotein receptor, the rate-limiting enzyme concentration that is used for the conversion of cholesterol bile acid, lipoprotein sex.The factor of the homeostasis of the blood cholesterol levels concentration that influence is human comprises the use of incidence rate, body movement and the various pharmaceutical agents of diet composition, smoking.The parameter of diet comprises the value volume and range of product of fat (saturated with polyunsaturated fatty acid), the quantity of cholesterol, and perhaps the value volume and range of product of fiber also has vitamin, for example vitamin C and D, and mineral, for example quantity of calcium.

Epidemiological study has shown the negative correlation (Wilson et al. (1988) Arteriosclerosis 8:737-741) that high density lipoprotein (HDL) and apolipoprotein (apo) A-I level and atherosclerotic event take place.Demonstrated the atherosclerotic pathological changes of inhibition to the rabbit injection HDL that raises with atherogenic meals and formed (Badimon et al. (1990) J.Clin.Invest.85:1234-1241).

Because its anti-atherogenic character, people apo A-I has become the object of intensive research.Interchangeable apolipoprotein comprises apoA-I, has lipid binding structural domain (Brouilletteand Anantharamaiah (1995) Biochim.Biophys.Acta 1256:103-129; Segrest et al. (1974) FEBS Lett.38::247-253).Supposed that Apo A-I has eight placed in-line repetition 22mer sequences, wherein great majority have the potentiality (Segrest et al. (1974) FEBS Lett.38::247-253) that form category-A amphiphilic helical structure.The feature of category-A amphiphilic spiral be included in polar-nonpolar at the interface the positively charged residue existence and at (Segrestet al. (1974) the FEBS Lett.38:247-253 that exists of the electronegative residue in the center of polar surface; Segrest et al. (1990) Proteins:Structure, Function, and Genetics 8:103-117).Shown that Apo A-I is cross-linked to form complex with phospholipid consumingly and promotes cholesterol to flow out from the cell that is rich in cholesterol.Alleviate effectively atherosclerotic one or more symptoms apo A-I serum levels release and keep, be proved to be unintelligible up to now.

Summary of the invention

It is right to the invention provides new peptide and aminoacid, use them and alleviated atherosclerosis and other diseases associated with inflammation, for example, rheumatoid arthritis, lupus erythematosus, polyarteritis nodosa, osteoporosis, Alzheimer, congestive heart failure, endothelial function disturbance, virus disease be one or more symptoms of the disease of A type influenza and for example multiple sclerosis for example.In some embodiments; discovery of the present invention is; when the time containing category-A amphiphilic spiral with the preparation of " D " amino acid residue and/or having the peptide of shielded amino and carboxyl terminal; organism can be administered to orally; being easy to be ingested and be discharged into serum, is effective for alleviating atherosclerotic one or more symptoms.In some embodiments, described peptide can and be still effectively with all " L " amino acid residue preparations, particularly when using by the approach except that Orally administered.

Also have a discovery to be, as described here have hydrophobic end amino acid or handle by one or more hydrophobic blocking groupses after have hydrophobic end amino acid, with have the tart and/or alkaline of inside as herein described and/or aliphatic and/or aromatic amino acid whose " little " peptide (for example, about three aminoacid of length range are to about 11 aminoacid) also can be alleviated atherosclerosis or be one or more symptoms of other pathological changes of feature with the inflammatory reaction.

Peptide of the present invention and/or aminoacid to usually for before stimulating-formation and the circulation of β high density lipoprotein like-particles (pre-beta high density lipopretein-like particles), and/or promote lipid transfer and detoxifcation, be effective.

Peptide described here and/or aminoacid also are effective to one or more symptoms for prevention of osteoporosis onset or inhibition or elimination osteoporosis.

Also have a surprised discovery to be, described peptide and/or aminoacid (for example strengthen being used to, enhancing synergistically) activity of inhibin (statin) and/or ezetimibe (Ezetimibe) or other cholesterol picked-up mortifier, thereby allow effectively to use inhibin or cholesterol picked-up mortifier, and/or inhibin or cholesterol picked-up mortifier are significantly strengthened at any given dose anti-inflammatory that becomes with low dosage more.

In some embodiments, the invention provides and (for example improve struvite situation, atherosclerosis, rheumatoid arthritis, lupus erythematosus, polyarteritis nodosa, osteoporosis, Alzheimer, virus disease, asthma, diabetes or the like) the peptide of one or more symptoms, or the combination of peptide and/or aminoacid are right.Some preferred peptide length range from 3 to about 5 aminoacid; Can be dissolved in ethyl acetate with concentration greater than about 4mg/mL; Dissolve in the aqueous buffer solution of pH 7.0; When aqueous environments contacts with phospholipid, form the particle of the about 7.5nm of diameter, and/or form stacked bilayer (stacked bilayer), bilayer size general (on the order of) 3.4 to 4.1nm, the about 2nm in interval between stacked (stack) middle bilayer; Have and be lower than about 900 daltonian molecular weight; To urge struvite (pro-inflammatory) HDL and change into anti-inflammatory HDL, or make anti-inflammatory HDL anti-inflammatory stronger; Do not have aminoacid sequence Lys-Arg-Asp-Ser (SEQID NO:238), wherein Lys-Arg-Asp and Ser are L aminoacid.In some embodiments; these peptide protection phospholipid (for example; 1-palmityl-2-arachidonic acyl group-sn-glyceryl-3-phosphocholine (PAPC), 1-stearoyl-2-arachidonic acyl group-sn-glyceryl-3-phosphocholine (SAPC)) and 1-stearoyl-2-arachidonic acyl group-sn-glyceryl-3-phosphoethanolamine (SAPE).In some embodiments, these peptides can be including, but not limited to any little peptide described here.

In some embodiments, the invention provides and (for example improve struvite situation, atherosclerosis, rheumatoid arthritis, lupus erythematosus, polyarteritis nodosa, osteoporosis, Alzheimer, virus disease, asthma, diabetes or the like) the peptide of one or more symptoms, or the combination of peptide and/or aminoacid are right.The feature of some preferred peptide is a formula: X ¹-X ²-X ³ _n-X ⁴, wherein n is 0 or 1; X ¹Be hydrophobic amino acid and/or have hydrophobic blocking group; X ⁴Be hydrophobic amino acid and/or have hydrophobic blocking group; With, when n is 0, X ²Be the aminoacid that is selected from the group that constitutes by acidic amino acid, basic amino acid and histidine; With, when n is 1, X ²And X ³Be acidic amino acid, basic amino acid, aliphatic amino acid or aromatic amino acid independently, thereby work as X ²When being acidic amino acid; X ³Be basic amino acid, aliphatic amino acid or aromatic amino acid; Work as X ²When being basic amino acid; X ³Be acidic amino acid, aliphatic amino acid or aromatic amino acid; With work as X ²When being aliphatic or aromatic aminoacid, X ³Be acidic amino acid, or basic amino acid.Some preferred peptide will be urged struvite HDL and change into anti-inflammatory HDL or make anti-inflammatory HDL anti-inflammatory stronger.In some embodiments, described peptide does not have aminoacid sequence Lys-Arg-Asp-Ser (SEQID NO:238), and wherein Lys, Arg, Asp and Ser are L aminoacid.Peptide of the present invention comprises the peptide according to following formula, and/or comprises the peptide of the peptide of following formula and/or the concatemer of this peptide (concatamers).

In some embodiments, X ¹And X ⁴Be independently selected from by alanine (Ala); valine (Val); leucine (Leu); isoleucine (Ile); proline (Pro); phenylalanine (Phe); tryptophan (Trp); methionine (Met); the serine (Ser) that has hydrophobic blocking group; the betanaphthyl alanine; the Alpha-Naphthyl alanine; nor-leucine; Cyclohexylalanine; the threonine (Thr) that has hydrophobic blocking group; the tyrosine (Tyr) that has hydrophobic blocking group; the lysine (Lys) that has hydrophobic blocking group; the arginine (Arg) that has hydrophobic blocking group; the ornithine (Orn) that has hydrophobic blocking group; the aspartic acid (Asp) that has hydrophobic blocking group; have the cysteine (Cys) of hydrophobic blocking group and have the group that the glutamic acid (Glu) of hydrophobic blocking group constitutes.

In some embodiments, peptide is trisome (tri-mer) (that is, N is 0).In some trisome, X ¹Be Glu, Leu, Lys, Orn, Phe, Trp or norLeu; X ²Be tart (for example, aspartic acid, glutamic acid, or the like), or alkalescence (for example, lysine, arginine, histidine, or the like), and X ⁴Be Ser, Thr, Ile, Leu, Trp, Tyr, Phe or norleu.In some embodiments, described peptide comprises the aminoacid sequence of peptide listed in the table 3.In some embodiments, described peptide is a shielded trisome as shown in table 3.

In some embodiments, n is 1, and described peptide is limbs (tetramer), or comprises limbs, wherein X ²And X ³Be acidic amino acid or basic amino acid independently, work as X like this ²When being acidic amino acid, X ³Be basic amino acid and work as X ²When being basic amino acid, X ³Be acidic amino acid, X ¹And X ⁴Can comprise selected aminoacid independently, for example, as noted above.In some embodiments, X ²And X ³Be independently selected from Asp, Glu, Lys, Arg and His.In some embodiments, described peptide comprises the aminoacid sequence of peptide listed in the table 4.In some embodiments, described peptide is shielded limbs as shown in table 4.

In another embodiment, n is 1, and described peptide is limbs, or comprises limbs, wherein X ²And X ³Be acidity, alkalescence or aliphatic amino acid independently, X ²Or X ³One of be acidity or basic amino acid, work as X like this ²When being acidity or basic amino acid, X ³It is aliphatic amino acid; With work as X ³When being acidity or basic amino acid, X ²It is aliphatic amino acid.X ¹And X ⁴Can comprise selected aminoacid independently, for example, as noted above.In some embodiments, X ²And X ³Be independently selected from the group that constitutes by Asp, Glu, Lys, Arg, His and Ile, the preferred group that constitutes by Asp, Arg, Leu and Glu that is selected from.In some embodiments, described peptide comprises the aminoacid sequence of peptide listed in the table 5.In some embodiments, described peptide is shielded limbs as shown in table 5.

In another embodiment, n is 1, and described peptide is limbs, or comprises limbs, wherein X ²And X ³Be acidity, alkalescence or aromatic amino acid independently, X ²Or X ³One of be acidity or basic amino acid, work as X like this ²When being acidity or basic amino acid, X ³It is aromatic amino acid; With work as X ³When being acidity or basic amino acid, X ²It is aromatic amino acid.X ¹And X ⁴Can comprise selected aminoacid independently, for example, as noted above.In some embodiments, X ²And X ³Be independently selected from the group that constitutes by Asp, Arg, Glu, Trp, Tyr, Phe and Lys.In some embodiments, described peptide comprises the aminoacid sequence of peptide listed in the table 6.In some embodiments, described peptide is shielded limbs as shown in table 6.

The present invention also provides peptide, and described peptide is five body constituents or comprises five body constituents (pentamer) (5-mer) that described five body constituents is with formula: X ¹-X ²-X ³-X ⁴-X ⁵Be feature, wherein X ¹Be hydrophobic amino acid and/or have hydrophobic blocking group; X ⁵Be hydrophobic amino acid and/or have hydrophobic blocking group; X ², X ³And X ⁴Be independently selected from aromatic amino acid or histidine; To urge struvite HDL with described peptide and change into anti-inflammatory HDL, or make anti-inflammatory HDL anti-inflammatory stronger.In some embodiments, X ¹And X ⁵Be independently selected from by alanine (Ala); valine (Val); leucine (Leu); isoleucine (Ile); proline (Pro); phenylalanine (Phe); tryptophan (Trp); methionine (Met); phenylalanine (Phe); tryptophan (Trp); methionine (Met); the serine (Ser) that has hydrophobic blocking group; the betanaphthyl alanine; the Alpha-Naphthyl alanine; nor-leucine; Cyclohexylalanine; the threonine (Thr) that has hydrophobic blocking group; the tyrosine (Tyr) that has hydrophobic blocking group; the lysine (Lys) that has hydrophobic blocking group; the arginine (Arg) that has hydrophobic blocking group; the ornithine (Om) that has hydrophobic blocking group; the aspartic acid (Asp) that has hydrophobic blocking group; have the cysteine (Cys) of hydrophobic blocking group and have the group that the glutamic acid (Glu) of hydrophobic blocking group constitutes.In some embodiments, X ², X ³And X ⁴Be independently selected from the group that constitutes by Phe, Val, Trp, Tyr and His.In some embodiments, described peptide comprises the aminoacid sequence of peptide listed in the table 7.In some embodiments, described peptide is shielded limbs as shown in table 7.

The present invention also provides the bigger peptide of one or more symptoms of improving struvite situation.In some embodiments, from 5 to 11 aminoacid of described peptide length range; End amino acid is hydrophobic amino acid and/or has hydrophobic blocking group; Non-end amino acid forms at least one acid domain and at least one basic domain; Described peptide will be urged struvite HDL and change into anti-inflammatory HDL, or make anti-inflammatory HDL anti-inflammatory stronger.

In some embodiments, from 5 to 11 aminoacid of described peptide length range; End amino acid is hydrophobic amino acid and/or has hydrophobic blocking group; Non-end amino acid forms at least one acid domain or a basic domain and at least one aliphatic structure territory; Described peptide will be urged struvite HDL and change into anti-inflammatory HDL, or make anti-inflammatory HDL anti-inflammatory stronger.

In other embodiments, from 5 to 11 aminoacid of described peptide length range; End amino acid is hydrophobic amino acid and/or has hydrophobic blocking group; Non-end amino acid forms at least one acid domain or a basic domain and at least one aromatic structure territory; Described peptide will be urged struvite HDL and change into anti-inflammatory HDL, or make anti-inflammatory HDL anti-inflammatory stronger.

In other embodiment, from 6 to 11 aminoacid of described peptide length range; End amino acid is hydrophobic amino acid and/or has hydrophobic blocking group; Two or more aromatic structures territory that non-end amino acid forms at least one aromatic structure territory or separates by one or more histidine; Described peptide will be urged struvite HDL and change into anti-inflammatory HDL, or make anti-inflammatory HDL anti-inflammatory stronger.

The present invention also provides one or more symptoms of improving struvite situation and the peptide that comprises one or more amphiphilic spirals.Thereby, the present invention includes the concatemer of peptide or peptide, its length range about 10 to about 30 aminoacid, preferred about 18 to about 30 aminoacid; Comprise at least one category-A amphiphilic spiral; Center at the non-polar plane of described amphiphilic spiral comprises one or more aliphatic or aromatic amino acid; Protection phospholipid is to the oxidation of antioxidant; And it is not the D-18A peptide.In some embodiments, described peptide comprises the aminoacid sequence of peptide listed in table 2 or the table 12.In some embodiments, described peptide is the shielded limbs as shown in table 2 or table 12.

In some embodiments; peptide protection phospholipid of the present invention (for example; 1-palmityl-2-arachidonic acyl group-sn-glyceryl-3-phosphocholine (PAPC); 1-stearoyl-2-arachidonic acyl group-sn-glyceryl-3-phosphocholine (SAPC)), 1-stearoyl-2-arachidonic acyl group-sn-glyceryl-3-phosphoethanolamine (SAPE) to antioxidant (for example; 13 (S)-HPODE, 15 (S)-HPETE, HPODE, HPETE, HODE, HETE, or the like) oxidation.

Any peptide described here can be at amino terminal amino acid (for example, X ¹) and/or carboxyl terminal aminoacid (for example, X ⁴, X ⁵, or the like) on have one or more hydrophobic blocking groups.Described blocking group can be attached to amino or carboxyl terminal and/or amino acid whose side chain (R group).Directly coupling of described blocking group (for example, via covalent bond) or coupling indirectly (for example, via joint).Preferred hydrophobic blocking group comprises; but be not limited to; tertbutyloxycarbonyl (Boc); Fmoc; nicotinoyl; OtBu; benzoyl; acetyl group (Ac); carbobenzoxy group; methyl ester; ethyl ester; propyl ester; butyl ester; pentyl ester; own ester; N-methylamino o-tolyl (N-methyl anthranilyl); 3 to 20 carbon alkyl; amide; 3 to 20 carbon hydrocarbyl groups; 9-fluorenes acetyl group (fluoreneacetyl) group; 1-fluorenes carboxyl (fluorenecarboxylic) group; 9-fluorenes carboxylic group; 9-Fluorenone-1-carboxyl (9-fluorenone-1-carboxylic) group; benzyloxycarbonyl (benzyloxycarbonyl; be also referred to as carbobenzoxy group as described above); xanthyl (Xan); trityl (Trt); 4-methyl trityl (Mtt); 4-methoxyl group trityl (Mmt); 4-methoxyl group-2; 3; 6-trimethyl-benzenesulfonyl (Mtr); mesitylene-2-sulfonyl (Mts); 4; 4-dimethoxy benzhydryl (Mbh); tosyl (Tos); 2; 2; 5; 7; 8-pentamethyl chromane-6-sulfonyl (2; 2; 5; 7; 8-pentamethyl chroman-6-sulphonyl) (Pmc); 4-methylbenzyl (MeBzl); 4-methoxybenzyl (MeOBzl); benzyloxy (BzlO); benzyl (Bzl); benzoyl (Bz); 3-nitro-2-pyridine sulfinyl (Npys); 1-(4; 4-dimethyl-2; 6-dioxy ring caproic subunit) ethyl (Dde); 2; 6-dichloro-benzenes methyl (2; 6-DiCl-Bzl); 2-chlorine benzyloxycarbonyl (2-Cl-Z); 2-bromo-benzyloxy-carbonyl (2-Br-Z); benzyloxymethyl (Bom); cyclohexyloxy (cHxO); tert-butoxy methyl (Bum); tert-butoxy (tBuO); the tert-butyl group (tBu); trifluoroacetyl group (TFA); 4-[N-{1-(4; 4-dimethyl-2; 6-dioxy ring caproic subunit)-3-methyl dibutyl)-amino } benzyl esters (ODmab); α-allyl ester (OAll); 2-propyloxy phenyl base ester (2-PhiPr); 1-[4; 4-dimethyl-2; 6-dioxy hexamethylene-1-base-subunit) ethyl (Dde), or the like.In some embodiments, described hydrophobic blocking group is selected from the group that is made of Boc, Fmoc, nicotinoyl and OtBu.In some embodiments, the blocking group sealing of the group of the N-of peptide terminal selected free Boc-, Fmoc-and nicotinoyl-formation, and/or the blocking group sealing of the group of terminal selected free tBu of the C-of peptide and OtBu formation.

Randomly, described peptide also can comprise at least one D aminoacid.In some embodiments, described peptide comprises a plurality of D-aminoacid, or even can comprise it all being D-aminoacid.In some embodiments, described peptide comprises alternative D-and L-aminoacid.Described peptide also can all be the aminoacid of L-form.Described peptide can be isolating (for example, pure basically), exsiccant or in solution, and/or goes up acceptable excipient composition with the pharmacology.In some embodiments, described peptide be suitable for going up acceptable mixed with excipients to the Orally administered pharmacology of mammal (for example, people or non-human mammal).Described peptide can be used as the unit formulation in the pharmaceutically acceptable excipient and/or provides as time release formulation.

Described peptide also can with one or more biotin couplings (for example, directly, via joint and/or via amino acid side chain).In some embodiments, biotin and lysine (Lys) coupling.

In some embodiments, it is right that the present invention also provides the aminoacid of one or more symptoms of improving struvite situation.Described aminoacid is to usually comprising first aminoacid that has at least one blocking group; Second aminoacid that has at least one blocking group; Wherein said first aminoacid and second aminoacid are that inhomogeneous aminoacid and wherein said aminoacid change into anti-inflammatory HDL to urging struvite HDL, or make anti-inflammatory HDL anti-inflammatory stronger.In the right various embodiments of aminoacid,, form the particle of the about 7.5nm of diameter and form stacked bilayer, bilayer size general 3.4 to 4.1nm, the about 2nm in the interval in stacked between the bilayer when when aqueous environments contacts with phospholipid.In some embodiments, described first and second aminoacid are independently selected from the group that is made of acidic amino acid, basic amino acid and nonpolar amino acid.In some embodiments, described first aminoacid is that tart or alkaline and described second aminoacid are nonpolar, or described first aminoacid is that nonpolar and described second aminoacid is tart or alkaline.In some embodiments, two aminoacid all make tart or alkaline.Randomly, described first and second aminoacid can covalently be coupled at together, for example, and directly or via the joint coupling.In some embodiments, thus aminoacid connect to form dipeptides via peptide bond.In some embodiments, described first aminoacid and second aminoacid are mixed together, rather than covalently bound.Described blocking group is including, but not limited to any protecting group described here.In some embodiments, the blocking group sealing of the group of sealing of the blocking group of the group of the selected free Boc-of described first aminoacid, Fmoc-and nicotinoyl-formation and the selected free tBu of described second aminoacid and OtBu formation.In some embodiments, each aminoacid has at least two blocking groups.In some embodiments, first blocking group of the group of the selected free Boc-of each aminoacid, Fmoc-and nicotinoyl-formation and be selected from second blocking group sealing of the group that constitutes by tBu and OtBu.In some embodiments, each aminoacid is by Boc and OtBu sealing.In various embodiments, described aminoacid is selected from the dipeptides of the group that is made of Phe-Arg, Glu-Leu and Arg-Glu to formation.In some embodiments, described aminoacid is selected from the dipeptides of the group that is made of Boc-Arg-OtBu, Boc-Glu-OtBu, Boc-Phe-Arg-OtBu, Boc-Glu-Leu-OtBu and Boc-Arg-Glu-OtBu to formation.

The present invention also provides and has comprised one or more and describe described peptide and/or aminoacid is right and the pharmaceutical preparation of pharmaceutically acceptable excipient at this.Usually, described peptide and/or aminoacid are to existing with effective dose.Described peptide and/or aminoacid are to also can be used as time release formulation and/or providing as unit dose formulations.In some embodiments, described preparation be configured to be used for Orally administered.In some embodiments, described preparation be configured to be used for by be selected from by Orally administered, (for example suck, nasal administration, oral cavity suck, or the like), the using of rectal administration, peritoneal injection, intravascular injection, subcutaneous injection, percutaneous, suck use, the approach of the group of formation such as intramuscular injection uses.

Test kit also is provided, described test kit comprises and contains one or more peptides described here and/or the right container of aminoacid, and guiding material, the pathological changes that is taught in treatment and with the inflammation is feature (for example, atherosclerosis, rheumatoid arthritis, lupus erythematosus, polyarteritis nodosa, asthma, osteoporosis, Alzheimer, virus disease, or the like) in use described peptide and/or aminoacid right.

The present invention also provides in mammal (people or non-human mammal) method of alleviating (for example, reducing or elimination) atherosclerotic one or more symptoms.It is right that this method usually comprises to one or more peptides described here and/or the aminoacid of described administration effective dose.Described peptide and/or aminoacid be to can using (for example, Orally administered) in pharmaceutically acceptable excipient, and randomly can unite (for example, before, afterwards or simultaneously) with lipid and use.Described use can comprise by be selected from by Orally administered, (for example suck, nasal administration, oral cavity suck, or the like), the approach of using the group that constitutes with intramuscular injection of rectal administration, peritoneal injection, intravascular injection, subcutaneous injection, percutaneous use described peptide and/or aminoacid right.In some embodiments, described mammal is to be diagnosed as the mammal with one or more atherosclerosis shapes.In some embodiments, described mammal is to be diagnosed as the mammal that has apoplexy or atherosclerotic risk.

In another embodiment, the invention provides the amelioration of inflammation sexually transmitted disease (STD) (for example becomes, atherosclerosis, rheumatoid arthritis, lupus erythematosus, polyarteritis nodosa, osteoporosis, multiple sclerosis, diabetes, asthma, Alzheimer, virus disease, or the like) one or more symptom methods.It is right that this method usually comprises to one or more peptides described here and/or the aminoacid of administration effective dose.Described peptide and/or aminoacid be to can using (for example, Orally administered) in pharmaceutically acceptable excipient, and randomly can unite (for example, before, afterwards or simultaneously) with lipid and use.Describedly use that the approach that can comprise the group that constitutes by use and the intramuscular injection that is selected from by Orally administered, suction, rectal administration, peritoneal injection, intravascular injection, subcutaneous injection, percutaneous is used described peptide and/or aminoacid is right.In some embodiments, described mammal is to be diagnosed as the mammal with one or more struvite pathological changes symptoms.In some embodiments, described mammal is the mammal that is diagnosed as the risk that has struvite pathological changes.

Described peptide of the present invention and/or aminoacid are to also (for example, ezetimibe (Ezetimibe) works synergistically with inhibin and/or with selectivity cholesterol picked-up mortifier.This method usually comprises one or more peptides described here of using effective dose with inhibin and/or cholesterol picked-up mortifier jointly.In some embodiments, described inhibin is selected from the group that is made of cerivastatin (cerivastatin), atorvastatin (atorvastatin), simvastatin (simvastatin), pravastatin (pravastatin), fluvastatin (fluvastatin), lovastatin (lovastatin), rosuvastatin (rosuvastatin) and Pitavastatin (pitavastatin).Described peptide can be before, use with inhibin and/or cholesterol picked-up mortifier afterwards or side by side.Described peptide and/or described inhibin and/or cholesterol picked-up mortifier can be used as unit dose formulations and use.In some embodiments, the described approach that comprises the group that constitutes by use and the intramuscular injection that is selected from by Orally administered, nasal administration, rectal administration, peritoneal injection, intravascular injection, subcutaneous injection, percutaneous of using is used described peptide and/or described inhibin.Described mammal including, but not limited to, be diagnosed as and have atherosclerotic one or more symptoms, or be diagnosed as the mammal that has apoplexy or atherosclerotic risk.

The present invention also provides the method for one or more symptoms that alleviation is relevant with atherosclerosis in mammal.This method usually comprises uses inhibin and/or selectivity cholesterol picked-up mortifier; Right with one or more peptides described here and/or the aminoacid of effective dose, wherein with at the effective dose that does not have the inhibin used under the right situation of described peptide and/or aminoacid or cholesterol picked-up mortifier compare, the effective dose of described inhibin and/or cholesterol picked-up mortifier is lower.In some embodiments, compare with the effective dose that the peptide used and/or aminoacid under the situation that does not have described inhibin and/or cholesterol picked-up mortifier are right, the right effective dose of described peptide and/or aminoacid is lower.In some embodiments, described inhibin is selected from the group that is made of cerivastatin, atorvastatin, simvastatin, pravastatin, fluvastatin, lovastatin, rosuvastatin and Pitavastatin.Described peptide can be before, use with inhibin and/or cholesterol picked-up mortifier afterwards or side by side.Described peptide and/or aminoacid are right, and/or described inhibin and/or cholesterol picked-up mortifier can be used as unit dose formulations and uses.In some embodiments, describedly use that the approach that comprises the group that constitutes by use and the intramuscular injection that is selected from by Orally administered, suction, rectal administration, peritoneal injection, intravascular injection, subcutaneous injection, percutaneous is used described peptide and/or aminoacid is right, and/or described inhibin.Mammal including, but not limited to, be diagnosed as and have atherosclerotic one or more symptoms, or be diagnosed as the mammal that has apoplexy or atherosclerotic risk.Described mammal including, but not limited to, be diagnosed as and have atherosclerotic one or more symptoms, or be diagnosed as the mammal that has apoplexy or atherosclerotic risk.

In another embodiment, the invention provides the method that in mammal, reduces or suppress one or more symptoms of osteoporosis.It is right that this method usually comprises to one or more peptides described here of described administration and/or aminoacid, and wherein said peptide and/or aminoacid are to using with enough reductions or the concentration of eliminating one or more symptoms of osteoporosis.In some embodiments, described peptide and/or aminoacid are to using with enough reductions or the concentration of eliminating the decalcification of skeleton.In some embodiments, described peptide and/or aminoacid are used the concentration with the recalcification (recalcification) of enough inducing skeleton.Described peptide and/or aminoacid to can with the pharmacology on can accept excipient (for example, being suitable for) combination to the Orally administered excipient of mammal.

In some embodiments, method of the present invention and/or peptide have been got rid of at WO97/36927 and/or United States Patent (USP) 6,037,323 and/or 6,376,464 and/or 6753,313 and/or at Garber et al. (1992) Arteriosclerosis and Thrombosis, disclosed any or multiple peptide among the 12:886-894.In some embodiments; the present invention has got rid of at United States Patent (USP) 4; 643,988 and/or in (1992) such as Garber disclosed any or multiple peptide, those peptides all are synthetic with D aminoacid when synthetic or peptide is blocking group by enantiotopic L aminoacid.In some embodiments, the present invention has got rid of and has had formula A ₁-B ₁-B ₂-C ₁-D-B ₃-B ₄-A ₂-C ₂-B ₅-B ₆-A ₃-C ₃-B ₇-C ₄-A ₄-B ₈-B ₉The peptide of (SEQ ID NO:(SEQID NO:1), wherein A ₁, A ₂, A ₃And A ₄Be aspartic acid or glutamic acid independently, or its homologue or analog; B ₁, B ₂, B ₃, B ₄, B ₅, B ₆, B ₇, B ₈And B ₉Be tryptophan, phenylalanine, alanine, leucine, tyrosine, isoleucine, valine or Alpha-Naphthyl alanine independently, or its homologue or analog; C ₁, C ₂, C ₃And C ₄Be that lysine or arginine and D are serine, threonine, alanine, glycine, histidine independently, or its homologue or analog; Condition is to work as A ₁And A ₂When being aspartic acid, A ₃And A ₄Be glutamic acid, B ₂And B ₉Be leucine, B ₃And B ₇Be phenylalanine, B ₄Be tyrosine, B ₅Be valine, B ₆, B ₈With D be alanine, and C ₁, C ₂, C ₃And C ₄Be lysine, B ₁It or not tryptophan.In some embodiments, though the present invention may get rid of aforesaid one or more peptides, SEQ ID NO:8 (4F or D4F) will clearly be included in the present invention.

In some embodiments, the present invention has got rid of any or multiple peptide and/or its D variant in WO 97/36927.Specific embodiment has been got rid of one or more following contents: as ApoA, apolipoprotein A-1, ApoA-2, ApoA 4, apolipoprotein B, apo B-48, Apolipoprotein B-100, apoC, apoC-1, apoC-2, apoC-3, ApoD, the apo E of describing in WO 97/36927.

In some embodiments, what also get rid of is at United States Patent (USP) 6,037, disclosed any or multiple peptide and/or its D variant in 323.Specific embodiment has been got rid of apo A-I agonist compounds, comprises (i) and has peptide or the peptide analogues that forms the amphiphilic alpha spiral under the situation of lipid and comprise 18 to 22 residues of following formula: Z ₁-X ₁-X ₂-X ₃-X ₄-X ₅-X ₆-X ₇-X ₈-X ₉-X ₁₀-X ₁₁-X ₁₂-X ₁₃-X ₁₄-X ₁₅-X ₁₆-X ₁₇-X ₁₈-Z ₂(SEQID NO:2), wherein X ₁Be Pro (P), Ala (A), Gly (G), Asn (N), Gln (Q) or D-Pro (p); X ₂It is aliphatic amino acid; X ₃Be Leu (L); X ₄It is acidic amino acid; X ₅Be Leu (L) or Phe (F); X ₆Be Leu (L) or Phe (F); X ₇It is basic amino acid; X ₈It is acidic amino acid; X ₉Be Leu (L) or Trp (W); X ₁₀Be Leu (L) or Trp (W); X ₁₁Be acidic amino acid or Asn (N); X ₁₂It is acidic amino acid; X ₁₃Be Leu (L), Trp (W) or Phe (F); X ₁₄Be basic amino acid or Leu (L); X ₁₅Be Gln (Q) or Asn (N); X ₁₆It is basic amino acid; X ₁₇Be Leu (L); X ₁₈It is basic amino acid; Z ₁Be H ₂N--or RC (O) NH--; Z ₂Be--C (O) NRR,--C (O) OR or--C (O) OH or its salt; Each R is independently--H, (C ₁-C ₆) alkyl, (C ₁-C ₆) thiazolinyl, (C ₁-C ₆) alkynyl, (C ₅-C ₂₀) aryl, (C ₆-C ₂₆) an alkane iso-aryl or 1 to 4 residue peptide or a peptide analogues that the iso-aryl (membered heteroaryl) that has of alkaryl, 5-20 or 6-26 have, wherein the one or more keys between residue 1-7 are the amide that replaces, the isostere or the amide analogies of amide independently; With at residue X ₁Up to X ₁₈Between each "-" indicate the amido link of amido link, replacement, the isostere or the amide analogies of amide independently; Or the (ii) form of the change of formula (I), wherein X ₁, X ₂, X ₃, X ₄, X ₅, X ₆, X ₇, X ₈, X ₉, X ₁₀, X ₁₁, X ₁₂, X ₁₃, X ₁₄, X ₁₅, X ₁₆, X ₁₇Or X ₁₈In the residue at least one conservatively replaced by another residue and/or its D variant.

In some embodiments, the present invention has got rid of the have sequence Lys-Arg-Asp-Ser peptide of (SEQ ID NO:238), in some embodiments, the present invention has got rid of the have sequence Lys-Arg-Asp-Ser peptide of (SEQ ID NO:238), and wherein Lys-Arg-Asp and Ser are L aminoacid.

In some embodiments, according to United States Patent (USP) 6,376, the detection method that provides in 464 is measured, and peptide of the present invention shown and be lower than 38%, preferably is lower than about 35%, preferredly to be lower than about 30% or be lower than about 25% LCAT activating activities.

Definition

Term " polypeptide ", " peptide " and " albumen " use interchangeably at this, refer to the polymer of amino acid residue.This term is applicable to such amino acid polymer, and one or more therein amino acid residues are amino acid whose artificial chemical analogs of corresponding natural generation, also is suitable for the amino acid polymer of natural generation.

Term " category-A amphiphilic spiral (class A amphipathic helix) " is meant the protein structure that forms alpha-helix, separating of polarization and non-polar plane, there is positively charged residue at the interface at polar-nonpolar, exist in the center of polar surface electronegative residue (referring to, Segrest et al. (1990) Proteins.:Structure for example, Function, and Genetics 8:103-117).

When relating to " improving atherosclerotic one or more symptoms ", term " improvement " is meant minimizing, prevention or eliminates with atherosclerosis and/or relevant diseases is one or more symptoms of feature.This reduction including, but not limited to, reducing or eliminating of the phospholipid of oxidation, atherosclerotic speckle forms and disruptive minimizing, the minimizing of for example heart attack of clinical events, angor or apoplexy, hypertensive decline, the synthetic decline of struvite protein biology, the minimizing of plasma cholesterol, or the like." improve atherosclerotic one or more symptoms " and also can refer to improve the blood flow that influenced by atherosclerosis to vascular bed.

Term " enantiomer aminoacid " is meant the aminoacid that exists with at least two kinds of forms of non-superimposable mirror image each other.Most of aminoacid (except that glycine) are enantiomorphous, exist with so-called L-type (L aminoacid) or D-type (D aminoacid).The aminoacid of most of natural generations is " L " aminoacid.Term " D aminoacid " and " L aminoacid " are used to refer to amino acid whose absolute configuration, rather than refer to the specific direction of rotation of plano-polarized light.Usage at this is consistent with those skilled in the art's normal usage.

Term " blocking group " is meant a kind of chemical group, and the character of this functional group is sealed or covered to (for example, side chain, α amino, α carboxyl, or the like) when it is attached to functional group on the aminoacid.Preferred amino terminal blocking group is including, but not limited to acetyl group or amino.Other amino terminal blocking group is including, but not limited to the alkyl chain in the fatty acid, propiono, formyl, or the like.Preferred carboxyl terminal blocking group is including, but not limited to the group that forms amide or ester.Term " side chain protected group " is meant the blocking group of protecting/seal amino acid whose side chain (that is R group).The side chain protected group is including, but not limited to amido protecting group, carboxy protective group and hydroxy-protective group for example nitro, tosyl of aryl ether and guanidine blocking group for example, or the like.

Term " protection phospholipid avoids the oxidation of oxidant " is meant; when phospholipid and oxidant (for example; hydrogen peroxide, 13-(S)-HPODE, 15-(S)-HPETE, HPODE, HPETE, HODE, HETE; or the like) when contacting, chemical compound reduces the ability of the oxidation rate (or quantity of the oxidized phospholipids that produces) of phospholipid.

Term " low density lipoprotein, LDL " or " LDL " are according to the definition of those skilled in the art's common usage.Usually, LDL is meant the lipid-protein complex that exists to d=1.063 with density range d=1.019 when separating with supercentrifugation.

Term " high density lipoprotein " or " HDL " are according to the definition of those skilled in the art's common usage.Usually, " LDL " is meant the lipid-protein complex that exists to d=1.21 with density range d=1.063 when separating with supercentrifugation.

Term " I organizes HDL " be meant reduce oxidation lipid (for example; in the low density lipoprotein, LDL) or the lipid of protection oxidation (for example avoid the high density lipoprotein of oxidized dose of oxidation or its composition; apoA-I, paraoxonase, platelet activating factor acetyl group hydrolytic enzyme, or the like).

Term " II organizes HDL " is meant at the protection lipid and avoids providing aspect the oxidation or at the lipid aspect of repairing (for example, reducing) oxidation reduce active or do not have active HDL.

Term " HDL composition " is meant the composition (for example, molecule) that comprises high density lipoprotein (HDL).The protection lipid avoids the detection of the HDL of oxidation or reparation (for example, reducing the lipid of oxidation), also comprises the detection (for example, apo A-I, paraoxonase, platelet activating factor acetyl group hydrolytic enzyme, or the like) of the composition that has shown this active HDL.

Term " people apo A-I peptide " is meant total length people apo A-I peptide, or comprises its fragment or the domain of category-A amphiphilic spiral.

As used herein " mononuclear cell reaction " be meant the monocytic activity that forms relevant " inflammatory reaction " feature with atheromatous plaque.The feature of mononuclear cell reaction is, mononuclear cell is attached to the cell (for example cell of blood vessel endothelium) of blood vessel wall, and/or chemotactic enters SE gap, and/or mononuclear cell is divided into macrophage, and/or the monocyte chemotaxis of in-vitro measurements (for example, utilizing neural detecting chamber).

When relating to the phospholipid quantity of oxidation, term " not variation " is meant there is not detectable variation, and preferred finger does not have (for example to change significantly on the statistics, at least 85%, preferred at least 90%, preferred at least 95% and most preferred at least 98% or 99% confidence level).Do not have detectable variation also can refer to a kind of detection, wherein the phospholipid level of oxidation has changed, but so much during when not having albumen described herein or with reference to other positive or negative contrast.

Abbreviation below this uses: PAPC:L-A-I-palmityl-2-arachidonic acyl group-sn-glyceryl-3-phosphocholine; POVPC:1-palmityl-2 (5-oxygen valeryl)-sn-glyceryl-3-phosphocholine; PGPC:1-palmityl-2-glutaryl-sn-glyceryl-3-phosphocholine; PEIPC:1-palmityl-2-(5, the different prostate alkane of 6-epoxy (epoxyisoprostane) E ₂)-sn-glyceryl-3-phosphocholine; ChC18:2: cholesteryl linoleic acid; ChC18:2-OOH: cholesteryl linoleic acid peroxidation hydrogen thing; DMPC:1, the two myristoyl myristoyls of 2--rac-glycerol-3-phosphocholine; PON: paraoxonase; HPF: standardized high power field; PON: paraoxonase; BL/6:C57BL/6J; C3H:C3H HeJ.

Using term " conservative replacements " about albumen or peptide is to embody the activity (specificity (for example, for lipoprotein)) that do not change molecule in fact or in conjunction with the aminoacid replacement of affinity (for example, for lipid or lipoprotein).Usually, conservative amino acid comprises an aminoacid is replaced with have similar chemical property another aminoacid of (for example, electric charge or hydrophobicity).Following six groups each self-contained be the aminoacid that typical conservative is replaced mutually: 1) alanine (A), serine (S), threonine (T); 2) aspartic acid (D), glutamic acid (E); 3) agedoite (N), glutamine (Q); 4) arginine (R), lysine (K); 5) isoleucine (I), leucine (L), methionine (M), valine (V); With 6) phenylalanine (F), tyrosine (Y), tryptophan (W).

Term " same " or " homogeneity " percentage ratio are meant in the context of two or more nucleic acid or peptide sequence, when carrying out maximum correspondence relatively or comparison, use the measurement of one of following sequence comparison algorithm or during by visual inspection, two or more sequences or subsequence are identical or the same amino acid residue or the nucleotide of particular percentile are arranged.For peptide of the present invention, on the total length of peptide, measure sequence homogeneity.

For sequence relatively, usually a sequence is as the reference sequence, and cycle tests compares with it.When using sequence comparison algorithm, will test with reference sequences being input in the computer, specify subsequence coordinate (coordinates), if necessary, specified sequence algorithm routine parameter.Sequence comparison algorithm calculates the sequence homogeneity percentage ratio of cycle tests with respect to reference sequences according to specified program parameter then.

Can pass through, for example, Smith ﹠amp; Waterman, local homology's algorithm of Adv.Appl.Math.2:482 (1981), Needleman ﹠amp; Wunsch, the homology alignment algorithm of J.Mol.Biol.48:443 (1970), Pearson ﹠amp; The similarity searching method of Lipman (1988) Proc.Natl.Acad.Sci.USA 85:2444, the computerization implementation of these algorithms (Wisconsin GeneticsSoftware Package, Genetics Computer Group, 575 Science Dr., Madison, GAP among the WI, BESTFIT, FASTA and TFASTA), or be used for sequence alignment comparison, best by visual inspection (generally, the same) referring to Ausubelet al..

An example of useful algorithm is PILEUP.That PILEUP uses is progressive, a plurality of sequence alignments from one group of relevant sequence are created in paired comparison, shows mutual relation and sequence homogeneity percentage ratio.It also marks and draws tree or tree diagram shows bunch mutual relation that is used to produce comparison.PILEUP has used Feng ﹠amp; The reduced form of the progressive comparison method of Doolittle (1987) J.Mol.Evol.35:351-360.This method of using is similar to Higgins ﹠amp; The method that Sharp (1989) CABIOS 5:151-153 describes.This program can be compared nearly 300 sequences, and the greatest length of each is 5,000 nucleotide or aminoacid.Multiple ratio starts from the paired comparison of two similar sequences to step, produce two by aligned sequences bunch.Then with this bunch and next maximally related sequence or bunch comparing by aligned sequences.The simple extension of the paired comparison by two independent sequences is compared two sequence clusters.By a series of progressive, paired comparisons, finish final aligning.By specifying concrete sequence and them to be used for the aminoacid in sequence zone relatively or nucleotide coordinate and coming working procedure by the designated program parameter.For example, reference sequences and other cycle tests relatively can be come to determine sequence homogeneity percentage ratio mutual relation, use following parameter: the terminal breach of the breach weight (3.00) of acquiescence, default gap length weight (0.10) and weighting.

Another example that is suitable for the algorithm of definite sequence homogeneity and sequence similarity percentage ratio is the BLAST algorithm, and it is described in Altschul et al. (1990) J.Mol.Biol.215:403-410.Being used to carry out the software that BLAST analyzes is that the public is obtainable in NCBI (http://www.ncbi.nlm.nih.gov/).This algorithm comprises, at first identifies high sub-sequence to (HSP) by the short word of identifying length W in search sequence, when with database sequence in during the word comparison of equal length, high sub-sequence is to coupling or satisfy some threshold value of just assessing and divide T.T is called as neighborhood word score threshold (Altschul et al., the same).These initial neighborhood word are sampled to have served as and are begun to search for the seed that contains their longer HSP with discovery.Both direction along each sequence extends described word sampling then, can be enhanced up to cumulative comparison score.For nucleotide sequence, the operation parameter M (reward score that the residue of coupling is right; Always＞0) and the N (point penalty of mispairing residue; Always＜0) calculate cumulative score.For aminoacid sequence, use the score matrix to calculate cumulative mark.When: cumulative comparison mark from its maximum that reaches numerical value X that descended; Because the accumulation of one or more negative mark residue comparisons, cumulative score reach zero or are lower than zero; Or arrive the ending of any one sequence; The time, stop at the extension of word sampling on each direction.The parameter W of BLAST algorithm, T and X have determined the sensitivity and the speed of comparison.What BLASTN program (for nucleotide sequence) acquiescence was used is word length (W) 11, expected value (E) 10, M=5, N=-4, relatively two chains.For aminoacid sequence, what BLASTP program acquiescence was used is, word length (W) 3, and expected value (E) 10 and BLOSUM62 score matrix are (referring to Henikoff ﹠amp; Henikoff (1989) Proc.Natl.Acad.Sci.USA 89:10915).

Except sequence of calculation homogeneity percent, the BLAST algorithm also carry out two between the sequence the similarity statistical analysis (referring to, for example, Karlin ﹠amp; Altschul (1993) Proc.Natl.Acad.Sci.USA, 90:5873-5787).A kind of similarity measurement method that the BLAST algorithm provides is minimum summation probability (P (N)), and it provides the probability that may occur accidentally mating between two nucleotide or aminoacid sequence to indicate.For example, if minimum summation probability is lower than approximately 0.1 with reference nucleic acid compare test nucleic acid the time, preferredly be lower than about 0.01 and most preferredly be lower than at about 0.001 o'clock, it is similar to reference sequences that nucleic acid is considered to.

Term " D-18A peptide " is meant to have sequence: the peptide of D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F (SEQ ID NO:3), wherein all enantiomer aminoacid is D type aminoacid.

For example, when using term " to use altogether " or when " using jointly " for peptide of the present invention and another active agent (for example, inhibin), thereby be meant and use described peptide and described active agent both can reach physiologic effect simultaneously.Yet these two kinds of reagent needn't be used simultaneously.In some embodiments, using of a reagent can using prior to another, yet, this using altogether causes that generally two reagent with the remarkable mark of the maximum serum-concentration of any given dose (for example, 20% or higher, preferred 30% or 40% or higher, preferred 50% or 60% or higher, most preferred 70% or 80% or 90% or higher) side by side is present in the body (for example, in the blood plasma).

When using term " detoxifcation ", be meant the lipid (oxidizing lipids) of the oxidation of removing some or all and/or the lipid (oxidized lipids) after the oxidation about lipid, LDL or HDL.Thereby, for example, picked-up all or some HPODE and/or HPETE (peroxide on the fatty acid) will stop or reduce these peroxide and enter LDL, thereby prevent or reduce the LDL oxidation.

Term " preceding-β high density lipoprotein like-particles (pre-beta high densitylipoprotein-like particles) " generally is meant the cholesterol that contains particle, described particle also contains apoA-I, it is littler, and with the lipid in most of HDL particles: protein rate is compared, and is the lipid poorness relatively.When with the FPLC separated plasma, these " preceding-β high density lipoprotein like-particles " are present in the FPLC fraction, and described FPLC fraction contains than the littler particle of particle in the main HDL peak, is arranged in the right side of FPLC chromatograph HDL, as shown in the related application USSN 10/423,830.

Term " oppositely lipid transfer and detoxifcation " is meant from tissue, for example remove lipid in the tremulous pulse, comprise cholesterol, other sterin, the phospholipid that comprises sterin, phospholipid, oxidant and the oxidation of oxidation, and the tissues around these transport out and arrive the organ, they can be detoxified and be secreted in described organ, for example be discharged in the bile and and be discharged in the urine by kidney by liver.Detoxifcation also refers to stop the phospholipid of oxidation as described in this to form, and/or disruptive oxidation phospholipid.

Term as used herein " biological sample " is meant from the organism of living or any sample of obtaining from dead organism.The example of biological sample comprises body fluid, tissue samples, cell and the cell line of taking from organism (for example people or non-human mammal).

Term when relating to hydrophobic blocking group or hydrophobic blocking groups " amide " comprises the monoamides of methyl amine or ethylamine.This term also comprises for example CO-NH-R of alkylamide, and wherein R is methyl, ethyl, or the like (for example, reaching about 7, preferred 9, preferred 11 or 13 carbon).

Term " D peptide " is meant a kind of peptide, and the wherein said one or more enantiomer aminoacid that comprise are D type aminoacid.In some embodiments, a plurality of enantiomer aminoacid are D type aminoacid.In some embodiments, the enantiomer aminoacid of half is D type aminoacid at least.In some embodiments, described peptide comprises alternative D and L type aminoacid.In some embodiments, all enantiomer aminoacid are D type aminoacid.

Term " L " peptide is meant a kind of peptide, and wherein all aminoacid (enantiomer aminoacid) is L type aminoacid.

The peptide of " will urge struvite HDL changes into anti-inflammatory HDL or make the anti-inflammatory HDL anti-inflammatory stronger " is meant, when (for example to mammal, the people, rat, mice, Deng) when using, the in vitro detection that maybe ought be used to be fit to (for example, as described here) time, (for example compare with the HDL in the control test, from having used than the control animal of the described peptide of low dosage or the HDL of detection, or from the negative control animal that lacks described peptide or the HDL of detection), HDL is changed into so a kind of HDL, the lipid oxidation of this HDL reduction or blocking oxide agent (for example, as at USSN 6,596, describe in 544), and/or have the paraoxonase activity of raising, and/or reduce the inductive monocyte chemotactic activity of LDL that produces by arterial wall cell.The preferably detectable variation of the change of HDL (from the protective effect that changes into of unprotect effect, or active aspect increase in protection).In preferred embodiment, described variation is to change significantly on the statistics, for example, (for example use any statistical test that is suitable for providing data set, t-test, amount of variability analysis (ANOVA), half parameter technology, nonparametric technique are (for example, Wilcoxon Mann-Whitney Test, Wilcoxon Signed RanksTest, Sign Test, Kruskal-Wallis Test, or the like)) determine.Preferably, the significant variation is at least 85%, preferred at least 90%, more preferred at least 95% and the significance of most preferred at least 98% or 99% confidence level on the statistics.In some embodiments, described variation is at least 10% variation, preferred at least 20% variation, preferred at least 50% variation and most preferred at least 90% variation.

When this uses, for example about use the aminoacid that comprises two seed amino acids to, about the combination of using peptide of the present invention, about with other pharmacological active agents (for example, one or more inhibin) together use peptide/aminoacid of the present invention right, or the like, two kinds of (or multiple) reagent are used in term " associating " expression, make at them to there being overlapping on some time sequencing at least aspect the physiologically active of organism.Thereby can be side by side and/or order use two or more reagent.The kind of using at order, as long as when second reagent of using is applied or becomes in vivo when activity is arranged, first reagent of using has been brought into play some physiological change to organism, before using second reagent even considerable delay (for example, several minutes and even a few hours or a couple of days) can be arranged.

Brief description of drawings

Accompanying drawing 1 has illustrated according to the synthetic synthetic schemes of the liquid phase of peptide of the present invention.

Accompanying drawing 2 has illustrated the process of the synthetic tetrapeptide of process that uses accompanying drawing 1 description.

Accompanying drawing 3 has shown the preincubate (pre-treatment) of Boc-Lys (Boc)-Arg-Asp-Ser (tBu)-OtBu (synthetic by D aminoacid entirely) (table 4 kind of SEQ ID NO:238) but has not been to hatch (Co-inc) altogether to have suppressed the inductive monocyte chemotactic activity of LDL that produced by human artery parietal cell (HAEC).With the peptide preincubate of cell and 125 μ g/mL, 250 μ g/mL or 500 μ g/mL, remove peptide then, added the LDL of the 100 μ g/mL cholesterol that have fresh culture, or together added the peptide of same concentrations with LDL, measure the monocyte chemotactic activity.

Accompanying drawing 4 has shown, the tetrapeptide of describing in the accompanying drawing 3 is added in the drinking water of apoE nude mouse, and is similar to D-4F, can be with the FPLC fraction behind HDL and the HDL from the short struvite anti-inflammatory that changes into.Described tetrapeptide or D-4F added to the concentration of 5 μ g/mL in the drinking water of mice (every kind of condition n=4) 18 hours.To the mice blood-letting, separate their lipoprotein by FPLC.The contrast people LDL of 100 μ g/mL cholesterol is added (LDL) or do not add (not adding) to human artery's wall coculture, or together add with HDL from normal human subject contrast experimenter's (+contrast HDL) 50 μ g/mL, or together add with HDL from the 50 μ g/mL of apoE nude mouse, described mice has been accepted the drinking water of no peptide (+water contrast HDL), or accepted tetrapeptide (+FPLC fraction (post-HDL FPLC fractions) D-TetraHDL) or behind D-4F (+D4F HDL) or the HDL, the FPLC fraction is from the apoE nude mouse of not accepting described peptide (behind+HDL water contrast) behind the described HDL, or contrasts the mice of LDL from the (+D-Tetra post HDL) mice of having accepted described tetrapeptide or the mankind that accepted 20 μ g/mLD-4F (+D4F post HDL) and 100 μ g/mL cholesterol.Detect the monocyte chemotactic activity of supernatant after 8 hours.

Accompanying drawing 5 shown, accepted D-tetrapeptide in the drinking water or the apoE nude mouse of D-4F and had the active LDL of the monocyte chemotactic of inducing still less.LDL from the FPLC fraction of the mice of description in the accompanying drawing 4 adds in the coculture with 100 μ g/mL.Detect the monocyte chemotactic activity of supernatant after 8 hours.

Accompanying drawing 6 has shown, orally give is entirely by the SEQ IDNO:258 (being called D-11 in the accompanying drawings) of the synthetic table 4 of D aminoacid, or D-4F has produced the HDL anti-inflammatory in the apoE nude mouse, but as among the D-4F, contain identical D aminoacid but with mixed and disorderly series arrangement, prevent the bonded peptide of lipid, do not produce described anti-inflammatory.Be instilled in the stomach of female 3 months big apoE nude mouses by flexible pipe by the synthetic five hectogamma SEQ IDNO:258 (D-11) of D aminoacid or 500 μ g D-4F (D-4F) or the mixed and disorderly D-4F (Scramb.Pept) of 500 μ g, (n=4), after 20 minutes (behind the tube feed 20 minutes) or after 6 hours (behind the tube feed 6 hours) mice is carried out blood-letting.Separated plasma separates HDL by FPLC.The culture of people's aortic endothelial cells has only been accepted culture medium (not having interpolation/detection contrast), do not contain (LDL/ detects contrast)) or contain the human HDL of standard control (the standard normal human subject LDL of the 100 μ gm/mL cholesterol of (LDL+ contrast HDL/ detects contrast) of 50 μ gm/mL cholesterol, or the mice HDL of the human LDL of the contrast of 100 μ gm/mL cholesterol and 50 μ gm/mL cholesterol together adds, and described mice HDL obtains oneself and accepted mixed and disorderly D-4F peptide (LDL+Scramb.Pept.HDL), or D-4F (LDL+D-4F HDL), or the mice of the SEQ ID NO:258 (LDL+D-11HDL) that entirely makes by D aminoacid.Hatched culture 8 hours.Detect the monocyte chemotactic activity of supernatant then.Numerical value is the meansigma methods ± SD of the monocytic number that moves in 9 high power fields. ^*Expression p＜0.001.

Accompanying drawing 7 shown, accepts D-4F or had by the apoE nude mouse of the SEQ IDNO:258 (being called D-11) (but not being from the mice of having accepted mixed and disorderly D-4F) of the synthetic table 4 of D aminoacid to induce the active LDL of lower monocyte chemotactic.LDL from the FPLC fraction of the mice of description in the accompanying drawing 6 adds in the culture with 100 μ g/mL.Detect the monocyte chemotactic activity of supernatant after 8 hours. ^*Expression p＜0.001, ^*Expression p＜0.01.

Accompanying drawing 8 shown, added by the SEQ ID NO:238 (being called D-1) (200 μ g/gm foods 18 hours) of the synthetic table 4 of D aminoacid afterwards at the food to the apoE nude mouse, and HDL is from the short struvite anti-inflammatory that is converted to.Detect contrast: additive-free, do not add to coculture; LDL adds the human LDL of standard control to coculture; + contrast HDL adds contrast normal human subject HDL to coculture.Food LDL (chow LDL) is from the LDL of the mice of only accepting food (chow); + Chow Autolog.HDL from the HDL of the mice of only accepting food, together adds with LDL from these mices; + D-1Autolog.HDL from the HDL of the mice of having accepted peptide, together adds in the coculture with LDL from these mices, measures the monocyte chemotactic activity.

Accompanying drawing 9 has shown that described tetrapeptide (the SEQ ID NO:258 in the table 4) ratio is effective ten times of external SEQID NO:238.In pre-incubation, described tetrapeptide is added with 100,50,25 or 12.5 μ gm/mL or does not add in the human artery parietal cell coculture, and hatches 2 hours.Wash culture then.Some reacting hole is only accepted culture medium (do not have and add) then.Other reacting hole has been accepted the standard normal human subject LDL (LDL) of 100 μ gm/mL cholesterol, or has together accepted this LDL (LDL+ contrasts HDL) with the human HDL of the standard control of 50 μ gm/mL cholesterol, hatches 8 hours.Detect the monocyte chemotactic activity of culture supernatants then.Numerical value is the meansigma methods ± SD of the monocytic number that moves in 9 high power fields.Accepted described tetrapeptide with above-mentioned concentration at 2 hours in the preincubate, the reacting hole that adds the LDL of 100 μ gm/mL cholesterol subsequently is specified (the LDL+ tetrapeptide is pressed μ gm/mL) in the drawings.

Accompanying drawing 10 has shown that the SEQ ID NOs:243,242 and 256 of table 4 (being called Seq No.5, Seq No.6 and Seq No.9 in the accompanying drawings) will change into anti-inflammatory HDL from the short struvite HDL of apoE nude mouse.Big female apo E nude mouse (at every turn handling n=4) fasting in two months 18 hours, with 20 μ gm peptides/Mus peritoneal injection L-tetrapeptide, or the pump pickle excipient.After two hours, under the slight anesthesia of different fluorine, collect blood from retro-orbital sinus.Separated plasma separates HDL by FPLC.Measure the character of struvite/anti-inflammatory of HDL then.The culture of people's aortic endothelial cells is only accepted culture medium (do not have and add), do not contain (LDL) or contain the standard normal human subject LDL of 100 μ gm/mL cholesterol of the human HDL of standard control (LDL+ contrasts HDL) of 50 μ gm/mL cholesterol, or the mice HDL of the human LDL of the standard control of 100 μ gm/mL cholesterol and 50 μ gm/mL cholesterol, described mice HDL is from the mice acquisition of having accepted described tetrapeptide or saline excipient (from the LDL+HDL of the mice of peritoneal injection).Hatched culture 8 hours.Detect the monocyte chemotactic activity of supernatant then.Numerical value is the meansigma methods ± SD of the monocytic number that moves in 9 high power fields.

Accompanying drawing 11 has shown that the SEQ ID NO:258 (being called S-11 in the accompanying drawings) of table 4 will change into anti-inflammatory HDL from the short struvite HDL of apoE nude mouse, than SEQ ID NO:254 and SEQ ID NO:282 better (being called S-7 and S-35 in the accompanying drawings).Big female apo E nude mouse (at every turn handling n=4) fasting in two months 18 hours, with 20 μ gm peptide/Mus peritoneal injection S-7 or S-11 or S-35, or pump pickle excipient (saline excipient).After two hours, under the slight anesthesia of different fluorine, collect blood from retro-orbital sinus.Separated plasma separates LDL and HDL by FPLC.Measure the character of struvite/anti-inflammatory of HDL then.The culture of people's aortic endothelial cells is only accepted culture medium (not having interpolation/detection contrast), do not contain (LDL/ detects contrast) or contain the standard normal human subject LDL of 100 μ gm/mL cholesterol of the human HDL of standard control (+contrast HDL/ detects contrast) of 50 μ gm/mL cholesterol, or the mice HDL of the mice LDL of 100 μ gm/mL cholesterol and 50 μ gm/mL cholesterol, it (is respectively LDL+S-7HDL that described mice HDL obtains from having accepted S-7 or S-11 or S-35, LDL+S-11HDL, LDL+S-35HDL) or the mice of saline excipient (LDL+ saline FDL).Hatched culture 8 hours.Detect the monocyte chemotactic activity of supernatant then.Numerical value is the meansigma methods ± SD of the monocytic number that moves in 9 high power fields. ^*p＜0.001。

Accompanying drawing 12.LDL from the FPLC fraction of the mice of description in the accompanying drawing 11 adds in the cell with 100 μ g/mL.Detect the monocyte chemotactic activity of supernatant after 8 hours.Detect contrast as described in the accompanying drawing 11.Saline LDL, the LDL of the mice of the saline excipient of using by oneself injection; S-7LDL comes the LDL of the mice that the SEQ ID NO:254 of table 4 personal as that describe in the accompanying drawing 11 injects; S-11LDL comes the LDL of the mice that the SEQ ID NO:258 of table 4 personal as that describe in the accompanying drawing 11 injects; S-35 comes the LDL of the mice that SEQ ID NO:282 personal as that describe in the accompanying drawing 9 injects.#p＜0.001。

Accompanying drawing 13 has shown serum amyloid A protein (SAA) blood plasma level behind the injection peptide.The SAA level of measuring in the blood plasma in 24 hours after the peptide of in injection accompanying drawing 11 and 12, describing. ^*p＜0.001。

Accompanying drawing 14 has shown that when the full L of using aminoacid synthesizes with oral administration, the SEQID NO:258 of table 4 will change into anti-inflammatory HDL from the short struvite HDL of apoE nude mouse.Female 3 months big apoE nude mouses (n=4) give the peptide of the SEQ IDNO:258 description of 200 μ g soluble in water such as table 4, and it is entirely by L aminoacid synthetic (being called S-11 in the accompanying drawings).Use described peptide or do not contain the water of peptide by stomach tube, give the mice blood-letting after 4 hours.Four mices of second group give standard Mus food powder type, that contain 200 microgram S-11, described S-11 is synthetic by L aminoacid entirely, add by per 1.0 gram powder Mus foods, to four mices in the cage altogether 4 gram powder Mus foods contain the described peptide of 800 micrograms altogether, or give the same powder Mus food that mice does not contain peptide.Allow mice obtain described food whole night, eaten up, give the mice blood-letting to the above food early.Separated plasma separates HDL by FPLC.Measure the character of struvite/anti-inflammatory of HDL then.The culture of people's aortic endothelial cells is only accepted culture medium (not having interpolation/detection contrast), do not contain (LDL/ detects contrast) or contain the standard normal human subject LDL of 100 μ gm/mL cholesterol of the human HDL of standard control (LDL+Cont.HDL/ detects contrast) of 50 μ gm/mL cholesterol, or the mice HDL of human LDL of the contrast of 100 μ gm/mL cholesterol and 50 μ gm/mL cholesterol, described mice HDL obtains from do not accept peptide (LDL+ does not have peptide HDL) by stomach tube (by the stomach gavage) or in mice food (meals of powder), or accept the mice of L-S-11 (LDL+L-S-11HDL).Hatched culture 8 hours.Detect the monocyte chemotactic activity of supernatant then.Numerical value is the meansigma methods ± SD of the monocytic number that moves in 9 high power fields.p＜0.001。

Accompanying drawing 15 shown, by L-aminoacid during synthetic and oral administration, L-S-11 has improved blood plasma paraoxonase (paraoxonase) activity when entirely.Blood plasma from the mice of describing in the accompanying drawing 14 is detected paraoxonase activity (the PON activity is shown as the unit of every 500ml blood plasma in the accompanying drawings).No peptide has only been accepted water or food, does not have the mice of peptide.L-S-11 describes as accompanying drawing 14, has given the mice of the peptide that the SEQ ID NO:256 of 200 microgram tables 4 describes in water or food.P＜0.001。

Accompanying drawing 16 has shown, when synthesizing with oral administration by D aminoacid entirely, the SEQID NO:238 (being called D-1) of table 4 and SEQ ID NO:258 (being called D-11) have caused the HDL anti-inflammatory in the apoE nude mouse, but when entirely by L aminoacid synthetic (L-1) and oral administration, SEQID NO:238 does not cause anti-inflammatory.Female 3 months big apoE nude mouse (n=4) give standard Mus powder type, that contain 0.5 milligram of every kind of peptide food, add by per 1.0 gram powder Mus foods, to four mices in the cage altogether 4 gram powder Mus foods contain 2 milligrams of described peptides altogether, or give the same powder Mus food that mice does not contain peptide.Allow mice obtain described food 24 hours, at this moment described food is eaten up, and gives the mice blood-letting.Separated plasma separates HDL by FPLC.The culture of people's aortic endothelial cells has only been accepted culture medium (not having interpolation/detection contrast), do not contain (LDL/ detects contrast) or contain the standard normal human subject LDL of 100 μ gm/mL cholesterol of the human HDL of standard control (LDL+ contrast HDL/ detects contrast) of 50 μ gm/mL cholesterol, or the mice HDL of human LDL of the contrast of 100 μ gm/mL cholesterol and 50 μ gm/mL cholesterol together adds, peptide (LDL+No Pep.HDL) is not accepted in described mice HDL acquisition certainly, or the full SEQ ID NO:238 (LDL+L-1HDL) that makes by L aminoacid of acceptance, or the SEQ ID NO:238 (LDL+D-1HDL) that entirely makes, or the mice of the SEQ ID NO:258 (LDL+D-11HDL) that makes by D aminoacid entirely by D aminoacid.Hatched culture 8 hours.Detect the monocyte chemotactic activity of supernatant then.Numerical value is the meansigma methods ± SD of the monocytic number that moves in 9 high power fields. ^*Expression p＜0.01 He ^*Expression p＜0.001.

Accompanying drawing 17 has shown, when synthesizing with oral administration by D aminoacid entirely, the SEQID NO:238 (D-1) of table 4 and SEQ ID NO:258 (D-11) caused the HDL anti-inflammatory and reduced the inductive monocyte chemotactic activity of LDL in the apoE nude mouse, and when not having during synthetic and oral administration by L aminoacid entirely.Mice separated plasma from accompanying drawing 16 is described separates HDL and LDL by FPLC.The culture of people's aortic endothelial cells has been accepted independent culture medium (not having interpolation/detection contrast), do not contain (LDL/ detects contrast) or contain the standard normal human subject LDL of 100 μ gm/mL cholesterol of the human HDL of standard control (LDL+ contrast HDL/ detects contrast) of 50 μ gm/mL cholesterol, 100 independent μ gm/mL cholesterol from body mice LDL (mLDL) or also have the mice HDL of 50 μ gm/mL cholesterol, peptide (mLDL+No Pep.HDL) is not accepted in described mice HDL acquisition certainly, or the full SEQ ID NO:238 (mLDL+L-1HDL) that makes by L aminoacid of acceptance, or the SEQ ID NO:238 (mLDL+D-1HDL) that entirely makes, or the mice of the SEQ ID NO:258 (mLDL+D-11HDL) that makes by D aminoacid entirely by D aminoacid.Hatched culture 8 hours.Detect the monocyte chemotactic activity of supernatant then.Numerical value is the meansigma methods ± SD of the monocytic number that moves in 9 high power fields. ^*Expression p＜0.05, ^*Expression p＜0.01 He ^{* *}Expression p＜0.001.

Accompanying drawing 18 has shown, when oral during to the mice administration, entirely by the SEQ ID NO:258 (D-11) of the synthetic table 4 of the D aminoacid HDL cholesterol concentration that raise, and give not have by L or the synthetic SEQ ID of D aminoacid NO:238 (being respectively L-1 or D-1) orally.Mensuration is from the blood plasma HDL cholesterol concentration of the mice of describing in accompanying drawing 16 and 17.No peptide HDL does not accept the blood plasma HDL cholesterol in the mice of peptide; L-1HDL is accepting by the blood plasma HDL cholesterol in the mice of the synthetic SEQ IDNO:238 of L aminoacid; D-1HDL is accepting by the blood plasma HDL cholesterol in the mice of the synthetic SEQ ID of D aminoacid NO:238; D-11HDL is accepting by the blood plasma HDL cholesterol in the mice of the synthetic SEQ ID of D aminoacid NO:258. ^*Expression p＜0.001.

Accompanying drawing 19 has shown, when oral during to the mice administration, entirely by the SEQ ID NO:258 (D-11) of the synthetic table 4 of D aminoacid HDL paraoxonase (PON) activity that raise, and orally give is not had by L or the synthetic SEQ ID of D aminoacid NO:238 (being respectively L-1 or D-1).Be determined at the paraoxonase activity among the HDL that describes in the accompanying drawing 18.Numerical value is the activity of per 500 microlitre blood plasma. ^*Expression p＜0.001.

Accompanying drawing 20 has shown that pravastatin and D-4F work synergistically and has reduced the large artery trunks pathological changes that described damage is to measure in the en of apoE nude mouse face prepared product.Five ages in week, female apoE nude mouse gave not have (the water contrast) added in their drinking water, pravastatin 50 μ g/mL, pravastatin 20 μ g/mL or D-4F 2 μ g/mL, or D-4F 5 μ g/mL, or pravastatin (PRAVA.) 20 μ g/mL add D-4F 2 μ g/mL, or pravastatin (PRAVA.) 50 μ g/mL add D-4F 5 μ g/mL.11 weeks back execution mice is measured pathological changes in en face large artery trunks prepared product.

Accompanying drawing 21 has shown that pravastatin and D-4F work synergistically and has reduced aortic sinus pathological changes in the apoE nude mouse.Five ages in week, female apoE nude mouse gave not have (the water contrast) added in their drinking water, pravastatin 50 μ g/mL, pravastatin 20 μ g/mL or D-4F 2 μ g/mL, or D-4F 5 μ g/mL, or pravastatin (P) 50 μ g/mL add D-4F 5 μ g/mL, or pravastatin (P) 20 μ g/mL add D-4F 2 μ g/mL.11 weeks back execution mice is measured the aortic sinus pathological changes.

Accompanying drawing 22 has shown that the SEQ ID NO:242 of D-4F and table 4 and SEQ ID NO:258 reduce the lipoprotein lipid peroxide in the apoE nude mouse significantly.The SEQ IDNO:242 of 50 μ g/gm (D-198 in the drawings) or SEQ ID NO:258 (D-203 in the drawings) or D-4F (entirely by the synthetic peptide of D aminoacid) are added in the food of apoE nude mouse, or mice is not having continuation nursing (do not have and add) on the food that adds.After 18 hours, give the mice blood-letting,, measure their low density lipoprotein, LDL (LDL) and the content of the lipid peroxide (LOOH) of high density lipoprotein (HDL) by the blood plasma fractionated of FPLC to them. ^*Expression p＜0.01.

Accompanying drawing 23 has shown the dissolubility of peptide in ethyl acetate.SEQ ID NO 254:Boc-Lys (ε Boc)-Glu-Arg-Ser (tBu)-OtBu; With SEQ ID NO 258:Boc-Lys (ε Boc)-Arg-Glu-Ser (tBu)-OtBu.What also show is the dissolubility of SEQ ID NO:250 in ethyl acetate.

Accompanying drawing 24 is when forming the 7.5nm particle with DMPC SEQ ID NO:258 when aqueous environments is mixed.1mg/mL DMPC suspension in phosphate buffered saline (PBS) (PBS) adds 10% dexycholate, dissolves up to DMPC.Add SEQ ID NO:258 or SEQ ID NO:254 (DMPC: peptide; 1: 10; Wt: wt), the dialysis reactant mixture.After the dialysis, remaining clarifying for SEQ ID NO:258 solution, is muddy for SEQ ID NO:254 solution after dexycholate is removed in dialysis still.Picture be with negative staining preparation and 147, the electron micrograph under the 420x amplification.Arrow has been pointed out SEQ ID NO:258 particle, is measured as 7.5nm (they look like little white particles).

Accompanying drawing 25, under aqueous environments, SEQ ID NO:258 is added to the particle (big is hollow) of the about 7.5nm of DMPC formation diameter, with stacked lipid peptide bilayer (big twill arrow) (pointing to the small arrow of the white line in disk cylindrical stacked), bilayer size general (on theorder of) 3.4 to 4.1nm, the about 2nm in the gap between the bilayer (black line in disk stacked between the white line).That describes in condition and enlargement ratio and the accompanying drawing 24 is identical.

Accompanying drawing 26 has shown that the peptide of the SEQ ID NO:258 that adds DMPC in aqueous environments to forms the loose structure (vesicularstructure) (white arrow) of stacked lipid-peptide bilayer (twill arrow) and about 38nm.

Accompanying drawing 27 has shown that the DMPC that does not have SEQ ID NO:258 in aqueous environments does not form the particle of the about 7.5nm of diameter, or stacked lipid-peptide bilayer, does not also form the loose structure of about 38nm.The DMPC vesicle that shows is 12.5-14nm.That describes in condition and enlargement ratio and the accompanying drawing 24 is identical.

Accompanying drawing 28 has shown the molecular model of the peptide of the SEQ ID NO:254 that compares with the peptide of SEQ ID NO:258.Red expression oxygen atom, blue expression nitrogen-atoms, grey colour specification carbon atom, white expression hydrogen atom.

Accompanying drawing 29 has shown the gap filling molecular model of the SEQ ID NO:254 that compares with SEQ ID NO:258.Arrow in this gap filling molecular model has been pointed out the polarity and the nonpolar part of this molecule.Identical in color-coded and the accompanying drawing 28.

Accompanying drawing 30 has illustrated the peptide trunk (in the picture of bottom) of the direction that provides for the top picture.

Accompanying drawing 31 has shown the molecular model of the SEQ ID NO:254 that compares with SEQ ID NO:258, has pointed out Ser (tBu)-OtBu group.Identical in color-coded and the accompanying drawing 28.

Accompanying drawing 32 has shown the molecular model of the SEQ ID NO:254 that compares with SEQ ID NO 258, has pointed out each blocking group.Identical in color-coded and the accompanying drawing 28.

Accompanying drawing 33 has shown that SEQ ID NO:258 (but not being SEQ ID NO:254) causes the naked HDL anti-inflammatory of apoE.

Accompanying drawing 34 has shown SEQ ID NO:258 rather than SEQ ID NO:254, reduces the aortic root atherosclerosis in the apoE nude mouse significantly.As the score of aortic root (aortic sinus) pathological changes in the mensuration apoE nude mouse of describing in the accompanying drawing 33.Mice number in every group shows (n=) in the bottom of image, shows the representative part of each group at the top of image.

Accompanying drawing 35 has shown SEQ ID NO:258 rather than SEQ ID NO:254, reduces the large artery trunks atherosclerosis in the en face prepared product in the apoE nude mouse significantly.As containing the percentage ratio on the large artery trunks surface of atherosclerotic lesion in the en face prepared product in the mensuration apoE nude mouse of describing in the accompanying drawing 33.Show the mice number (n=) in each group in the bottom in figure left side, show on the figure right side separately with food and raise, or with the representative aorta of the mice of the food raising that has replenished SEQ ID NO:258.

Accompanying drawing 36 has shown entirely and has reduced atherosclerosis significantly by the synthetic SEQ ID of L aminoacid NO:250.With food diet (Chow) or be supplemented with the ApoE nude mouse (20 every group) that 200 μ g/gm are kept by the food of the synthetic SEQ ID of L aminoacid NO:250 (250) entirely.12 weeks back execution mice, the large artery trunks surface area % of mensuration pathological changes in en face prepared product.*p＝0.012。

Detailed Description Of The Invention

The present invention relates to such discovery, design is with simulation category-A Amphiphilic helix motif (Segrestet al. (1990) Proteins.:Structure, Function, and Genetics 8:103-117) synthetic peptide can with phospholipids incorporate, show many biological properties that are similar to human apo-A-I albumen. Especially, discovery of the present invention is, when using D amino acid to prepare this peptide, this peptide demonstrates the serum half-life of remarkable rising and particularly when amino and/or carboxyl terminal are closed, even can use orally.

Also have a surprising discovery to be, these peptides can stimulate formation and the circulation of front-β HDL like-particles. In addition, described peptide can strengthen/effect of synergize statin, allows that inhibin uses with remarkable low dosage, or at any given dose significantly higher anti-inflammatory arranged. Find that also peptide described here can suppress and/or prevent and/or treat one or more symptoms of osteoporosis. Before described peptide also can improve-β HDL; And/or provide paraoxonase (paroxynase) activity of HDL.

In addition, a surprising discovery of the present invention is that this D type peptide has been preserved the BA of corresponding L-type peptide. Use the interior zooscopy of body of this D type peptide to show effective oral delivery, the ability of the serum half-life of rising and alleviation or atherosclerotic one or more symptoms of prevention/inhibition.

Also have a surprising discovery to be, some little peptide to forming by minimum two amino acid or single amino acid, preferably (but dispensable) one or more described amino acid are amino acid whose D-stereoisomers, with have hydrophobic domains to allow lipid protein to interact, with the hydrophilic-structure territory to allow to a certain degree water-soluble, also have significant antiinflammatory property. Be not limited to specific theory, it is believed that described peptide described here or amino acid pair and the phosphatide of short struvite oxidation to form required " seed molecule (seeding molecules) " for example Ox-PAPC, POVPC, PGPC and PEIPC are combined. Because many struvite situations are mediated by the lipid of oxidation at least in part, we believe, described peptide of the present invention or amino acid is being to being effectively improving aspect the described inflammatory conditions, the formation of the known or lipid oxide that results from BA under a cloud of described situation. These are including, but not limited to atherosclerotic, rheumatoid arthritis, lupus erythematosus, polyarteritis nodosa, multiple sclerosis, asthma, diabetes, Alzheimer's and osteoporosis. Described " little peptide " usually length range from 2 or 3 amino acid to about 15 amino acid, preferred from about 4 amino acid to about 10 or 11 amino acid, most preferred from about 4 to about 8 or 10 amino acid. Described peptide is usually to have hydrophobic end amino acid or to have been processed end amino acid as feature by adhering to one or more hydrophobic " protection " group by hydrophobization. At this internal structure of described peptide is described in more detail.

In addition, a surprising discovery of the present invention is, many physical propertys (are for example predicted little peptide of the present invention, be less than 10 amino acid, preferably be less than about 8 amino acid, preferred from about 2 or 3 to about 5 or 6 amino acid) or the right ability of amino acid so that the atherosclerotic the stronger and releasing mammal of HDL anti-inflammatory and/or other pathology take inflammatory reaction as feature. Described physical property is included in highly dissoluble (for example, greater than about 4mg/mL) and the dissolubility in the aqueous buffer solution of pH 7.0 in the ethyl acetate. At contact phosphatide, for example 1, during two myristoyls of 2--sn-glyceryl-3-phosphocholine (DMPC), in aqueous environments, especially effective little peptide forms the about 7.5nm of diameter (± 0.1nm) particle, and/or form stacked bilayer, bilayer size general 3.4 to 4.1nm, the about 2nm in gap between the bilayer in stacked, and/or also form the loose structure of about 38nm. Some preferred embodiment in, described little peptide or amino acid are to having the molecular weight that is lower than about 900Da.

I. stimulate formation and the circulation of front-β HDL like-particles

The accumulation aspect that reverse cholesterol transport is considered to be in the prevention lipid is important, and the accumulation of described lipid easily produces atherosclerotic (Shah et al. (2001) Circulation, 103:3047-3050). Many people believe that the lipid that is significant is cholesterol. Our laboratory is verified, and crucial lipid is the phosphatide of oxidation, causes inflammatory reaction (Navabet al. (2001) Arterioscler Thromb Vasc Biol., 21 (4): 481-488 in atherosclerotic; Van Lenten etal. (001) Trends Cardiovasc Med, 11:155-161; Navab M et al. (2001) Circulation, 104:2386-2387).

These inflammatory reaction also may be to cause the patch erosion of heart attack and apoplexy or the reason of breaking. The risk inverse correlation of HDL-cholesterol level and heart attack and apoplexy (Downset al. (1998) JAMA 279:1615-1622; Gordon et al. (1977) Am J Med., 62:707-714; Castelli et al. (1986) JAMA, 256:2835-2838).

Before-β HDL be generally considered to be promote the most activated HDL cut aspect the reverse cholesterol transport (for example, from surrounding tissue for example artery collect cholesterol, it is transported to liver is used for being discharged to bile; Referring to, Fielding and Fielding (2001) Biochim Biophys Acta, 1533 (3): 175-189). Yet, because front-β HDL loops the failure of ripe α-migration HDL, for example, LCAT defective or inhibition, the level of front-β HDL can be enhanced (O ' Connor et al. (1998) J Lipid Res, 39:670-678). In the coronary artery disease patient, reported high-caliber before-β HDL (Miida et al. (1996) Clin Chem., 42:1992-1995).

In addition, have been found that the male sex than the women have higher levels of before-β HDL, but the male sex's coronary heart disease risk is than women big (O ' Connor et al. (1998) J Lipid Res., 39:670-678). Thereby the static measurement of front-β HDL level itself is dispensable for the risk of prediction coronary artery disease. Yet cholesterol enters (Fielding and Fielding (2001) Biochim Biophys Acta, 1533 (3): 175-189) that ripe HDL generally is considered to atherosclerotic is had protectiveness via front-β HDL circulation. In addition, we are verified, play the protective effect of antagonism LDL oxidation except the lipid of deoxidation from arterial wall cell by this approach.

Although relatively low absorptivity is arranged when Orally administered, peptide of the present invention (D-4F) is highly active for example.

In the research of the Apo-E of Orally administered D-4F nude mouse, we have determined in intestinal absorption after 20 minutes, D-4F formed the apoA-I that contains relatively high quantity and paraoxonase little front-β HDL like-particles. In fact, estimate before these-apoA-I quantity in the β HDL like-particles according to the Western trace, with comparison apoA-I quantity and D-4F quantity (measuring by radioactivity or LC-MRM) in these particles, show that when D-4F absorbs it has served as the catalyst of the formation that causes these front-β HDL like-particles from intestines. The D-4F that derives in this a small amount of intestines seems, the apoA-I, paraoxonase and the cholesterol that have replenished some enter these particles, the order of magnitude surpassed D-4F quantity (referring to, for example, Navab et al. (2004) Circulation, 109:r120-r125).

Thereby, after absorbing, D-4F of the present invention and other peptide or amino acid to replenish rapidly relative a large amount of apoA-I and paraoxonase with before forming-β HDL like-particles, it very likely is the most potent particle that promotes reverse cholesterol transport and destroy the lipid of BA oxidation. We believe, (incorporation) mixed subsequently rapidly in ripe HDL in the formation of these particles and they, possible explanation our the atherosclerotic remarkable minimizings in ldl receptor nude mouse and apoE-nude mouse, observed, described ldl receptor nude mouse is the diet of Western mode, described apoE-nude mouse is the laboratory rodent chow diet, the variation of described minimizing and plasma cholesterol or HDL-cholesterol aspect have nothing to do (the same).

Thereby, in one embodiment, the invention provides by using one or more peptides described here or amino acid to stimulating the front-formation of β HDL like-particles and the method for circulation. Thereby described peptide or amino acid are to promoting lipid transfer and detoxifcation.

II. alleviate atherosclerotic symptom

We find, normal HDL is suppressed at three steps of formation aspect of the LDL of appropriate oxidation. In those researchs (referring to, co-pending application USSN 09/541,468, on March 31st, 2000 submitted to), we have proved, at external use A-I or apo A-I simulating peptide (37pA) handler LDL, from LDL, remove the seed molecule, comprised HPODE and HPETE. These seed molecules be human artery parietal cell coculture can oxidation LDL and LDL to induce arterial wall cell to produce monocyte chemotactic active required. We have also proved after apo A-I is injected into mouse or is infused into human body, from LDL oxidation to the human artery parietal cell behind injection/infusion apo A-I of mouse or human volunteer separation resistance is arranged, in the arterial wall cell coculture, do not induce the monocyte chemotactic activity.

The defencive function of some peptide of the present invention is in mother's application (09/645 of submission on August 24th, 2000; 454; 09/896 of submission on June 29 calendar year 2001; 841; WO 02/15923 (PCT/US01/26497) with submission on June 29 calendar year 2001) illustrated in; referring to, the accompanying drawing 1-5 among the WO 02/15923 for example. Accompanying drawing 1 among the WO 02/15923, A, B, C and D have shown in the ApoE nude mouse¹⁴C-D-5F is related with blood constituent. It has also shown, from feeding atherogenic diet and inject the HDL of the mouse of PBS, fails to suppress the oxidation of human LDL and fail to suppress the monocyte chemotactic activity that LDL induces in human arterial wall cell coculture. In contrast, inject atherogenic diet and every day the HDL of the mouse of peptide described herein from feeding, suppress human LDL oxidation and in coculture, prevent monocyte chemotactic that LDL induces active aspect, be the same effective (accompanying drawing 2A and 2B among the WO 02/15923) with normal human subject HDL. In addition, take from and feed atherogenic diet and inject the LDL of the mouse of PBS every day, and take from the LDL of mouse that feeds identical diet but inject the peptide 5F of 20 μ g every day and compare the easier oxidized and easier monocyte chemotactic activity of inducing. As if the D peptide do not have immunogenicity (accompanying drawing 4 among the WO 02/15923).

Human arterial wall cell with from feed atherogenic diet and injection according to the HDL of the mouse of peptide of the present invention and the vitro reactions of LDL, be consistent with the protective effect that shows in this peptide body. Although as T-CHOL percentage, T-CHOL, LDL-cholesterol, IDL+VLDL-cholesterol and lower HDL-cholesterol that similar level is arranged, the animal of having fed atherogenic diet and having injected described peptide has significantly lower pathology score (accompanying drawing 5 among the WO 02/15923). Thereby described peptide has stoped the progress of atherosclerotic lesion in the mouse of atherogenic diet of feeding.

Thereby, in one embodiment, the invention provides and improve and/or prevention of arterial is atherosis and/or the method for one or more symptoms of other situation take inflammatory reaction as feature.

III. alleviate the atherosclerotic symptom relevant with acute inflammatory response.

Peptide of the present invention or amino acid are to also being useful under many circumstances. For example, we observe, and cardiovascular complication (for example, atherosclerotic, apoplexy, etc.) often is accompanied by or is following the outbreak of acute stage inflammatory reaction. This acute stage inflammatory reaction usually with the inflammatory disease of recurrent (for example, leprosy, pulmonary tuberculosis, systemic lupus erythematosus and rheumatoid arthritis), virus infections (for example, influenza), bacterial infection, fungal infection, organ transplant, wound or other wound, the dummy of implantation, biofilm, etc. relevant.

Surprising discovery of the present invention is, uses one or more peptides described here, can reduce or prevent during acute phase response or the formation of the phosphatide of oxidation afterwards, thus alleviation or the elimination cardiovascular complication relevant with this situation.

Thereby for example, we are verified, and grippal result is the attenuating in HDL aspect the acetyl group hydrolytic enzyme activities of paraoxonase and platelet activation. Be not limited to specific theory, we believe, result as these HDL loss of enzyme activity, also as the related result who during acute phase response, helps oxidation protein with HDL, HDL no longer can stop the LDL oxidation, and no longer can stop the monocyte chemotactic activity of the endothelial cell that LDL induces.

We observe, (for example inject very low dose every day after infecting influenza A virus, for mouse 20 micrograms) the experimenter of polypeptide of the present invention in, the paraoxonase level does not reduce, and the generation of the phosphatide of the oxidation of BA surpasses background. This shows, other event that can maybe can produce the acute stage inflammatory reaction at influenza infection (for example, because virus infections, bacterial infection, wound, transplanting, various auto immune conditionses etc.) during, (for example use to the patient who suffers from known coronary artery disease, oral ground or by injection) D-4F (and/or other peptide of the present invention), thereby we can prevent the heart attack relevant with the pathology that produces this inflammatory states and the rising of apoplexy incidence by this short.

Thereby, in some embodiments, the present invention expection is used one or more peptides of the present invention or amino acid pair to the experimenter, there is the risk of acute inflammatory response in described experimenter or is bearing acute inflammatory reaction, and/or exists the risk of atherosclerosis shape or bearing atherosclerotic symptom.

Thereby, for example, in influenza season, suffer from coronary heart disease or have the people of coronary heart disease risk can prophylactically use polypeptide of the present invention or amino acid pair. Stand the struvite situation of recurrent, for example rheumatoid arthritis, various autoimmune disease, etc. people (or animal) can be with polypeptide treatment of the present invention, to alleviate or prevention of arterial is atherosis or the development of apoplexy. Stand wound (trauma), for example, acute injury, tissue transplantation, etc. people (or animal) can be with polypeptide of the present invention treatment, to alleviate or prevention of arterial is atherosis or the development of apoplexy.

In some cases, this method must have the generation of acute inflammatory response or the diagnosis of risk. Described acute inflammatory response usually comprises the change of metabolic in the liver and Gene regulation aspect. Except immune, cardiovascular and central nervous system, this relates to the dynamic homeostatic process of all Major Systems of health. Usually, acute phase response only continues a couple of days, yet, in the situation of chronic or recurrent inflammation, the unusual continuity of some aspect of acute phase response may promote to be accompanied by the potential tissue damage of described disease, also may cause further complication, for example angiocardiopathy or albumen precipitation disease amyloidosis for example.

The biosynthesis situation of the liver that an importance of described acute phase response is fundamentally changed. Under normal environment, liver is with the plasma protein of the synthetic peculiar scope of stable concentration. Many kinds in these albumen have important function, and the higher blood plasma level of these acute phase reactants (APR) or acute phase protein (APP) is required during struvite post-stimulatory acute phase response. Although most of APR are synthetic by liver cell, there are some to be produced by other cell type, comprise monocyte, endothelial cell, fibroblast and adipocyte. Most of APR be induced above normal level 50% between the several times. In contrast, main APR can increase above normal level 1000 times. This group comprises serum amyloid A protein (SAA) and human C reactive protein (CRP) or its homologue in mouse, SAP (SAP). So-called negative APR PC during acute phase response has reduced, to allow that liver is in the raising aspect the ability of the synthetic APR that induces.

In some embodiments, therefore assess described acute phase response or risk by measuring one or more APP. Assessing this mark is known for those skilled in the art, exists commercial company that this measurement is provided (for example, by Cardiotech Services, Louisville, KY measures AGP).

IV. the activity of synergize statin.

What also find is, the D-4F (1 μ g/mL) that adds low dosage in the drinking water of apoE nude mouse do not improve significantly in 24 hours the HDL function (referring to, for example, related application USSN10/423,830). In addition, the Atorvastatin or the Pravastatin that add individually 0.05mg/mL in the drinking water of apoE nude mouse did not improve the HDL function in 24 hours. Yet, when Atorvastatin or the Pravastatin with 0.05mg/mL together adds D-4F 1 μ g/ml in the drinking water to, in the HDL function aspects significant improvement has appearred. In fact, the short naked HDL of struvite apoE become normal human subject HDL with 350 μ g/ml the same have anti-inflammatory (h, HDL, referring to, for example, related application USSN10/423,830).

Thereby the dosage itself of independent D-4F or independent inhibin is to the not impact of HDL function, works synergistically when administration together. When together giving apo E nude mouse with D-4F and inhibin, in human arterial wall cell coculture, aspect the inflammatory reaction of preventing to be induced by the effect of HPODE oxidation PAPC, the short struvite HDL of the mouse of 50 μ g/mL HDL-cholesterol is the same effective with the normal human subject HDL of 350 μ g/mL HDL-cholesterol.

Thereby, the method that strengthens the activity of inhibin is provided In some embodiments of the present invention. Described method usually comprises with one or more inhibin to be used simultaneously such as one or more peptides described here or amino acid pair. Realized improving between inhibin and the oral peptide atherosclerotic synergy such as D-4F described here or other similar peptide. For this reason, inhibin can be used with significantly lower dosage, thereby avoid using relevant various harmful side effect (for example, muscle consumption) with the inhibin of high dose, and/or strengthened significantly at the antiinflammatory property of any given dose inhibin.

V. suppress/treat osteoporosis

Angiosteosis and osteoporosis usually coexist as (Ouchi et al. (1993) Ann NY Acad Sci., 676:297-307 among the same experimenter; Boukhris and Becker (`1972) JAMA, 219:1307-1311; Banks et al. (1994) Eur J Clin Invest., 24:813-817; Laroche etal. (1994) Clin Rheumatol., 13:611-614; Broulik and Kapitola (1993) EndocrRegul., 27:57-60; Frye et al. (1992) Bone Mine., 19:185-194; Barengolts et al. (1998) Calcif Tissue Int., 62:209-213; Burnett and Vasikaran (2002) Ann ClinBiochem.; 39:203-210.Parhami et al. (1997) Arterioscl Thromb Vasc Biol.; 17:680-687); show the LDL (MM-LDL) of mild oxidation and the BA lipid among the MM-LDL [namely; the 1-palmityl of oxidation-2-arachidonic acyl group-sn-glyceryl-3-phosphocholine) (Ox-PAPC)]; and different prostate alkane (isoprostane); the different PGE2 of 8-; rather than unoxidized phosphatide (PAPC) or the different Prostaglandin F2a of different prostate alkane 8-; vascular cell (CVC in external evoked alkaline phosphatase activities and calcification; calcifying vascular cells) osteoblastic differentiation has still suppressed the differentiation of MC3T3-E1 osteocyte.

Osteon is similar in appearance to arterial wall, because osteon is in the center of the inner chamber of endothelial cell arrangement, described inner chamber is centered on by the SE gap of containing matrix and fibroblast-like cell, and it centers on (the same) by the preosteoblast that has occupied the position that is similar to smooth muscle cell in arterial wall and Gegenbaur's cell conversely. Girder bone Gegenbaur's cell also with marrow endothelial under the gap form interface (the same). The supposition such as Parhami, lipoprotein can pass through the endothelium of bone artery, is deposited in the SE gap, and here they can be as experience oxidation (the same) in coronary artery. According to their vitro data, their prediction will cause osteoblast differentiation and the mineralising of reduction in the SES of bone artery and the LDL oxidation in marrow, and this will promote osteoporosis (the same). Their hypothesis is further predicted, as with CAC, the LDL level will become with osteoporosis positive correlation (Pohleet al. (2001) Circulation, 104:1927-1932), but the HDL level becomes negative correlation (Parhami et al. (1997) Arterioscl Thromb Vasc Biol., 17:680-687) with osteoporosis.

External, the osteoblast differentiation of marrow stromal cell M2-10B4 is suppressed by MM-LDL, but is not suppressed (Parhami et al. (1999) J Bone Miner Res., 14:2067-2078) by natural LDL. When the marrow stromal cell cultivated from atherosclerotic susceptible C57BL/6 (BL6) mouse of the low fat food diet of having fed, healthy and strong osteogenic differentiation (the same) appears. In contrast, during the marrow stromal cell of the mouse after higher fatty acid, atherogenic diet is taken from cultivation, they do not experience osteogenic differentiation (the same). This observed result is particular importance, because it provides the possible explanation (Nuttall and Gimble (2000) Bone, 27:177-184) of the skeletonization potentiality that reduce for marrow stromal cell in the development of osteoporosis. In vivo, the decline of skeletonization potentiality aspect is accompanied by adipogenic increase (the same) in the porotic bone.

It is found that, in the drinking water of apoE nude mouse, add the mineral density that has improved significantly trabecular bone in 6 weeks of D-4F, it is believed that in addition other peptide of the present invention also works similarly.

Our data show that osteoporosis can be considered to " atherosclerotic of bone ". This it seems the result of effect of the lipid that is oxidation. HDL has destroyed the lipid of these oxidations and has promoted osteoblast differentiation. Our data show, to administration peptide of the present invention (for example, in the drinking water of apoE nude mouse) only with increasing significantly trabecular bone once week.

This shows, peptide described here or amino acid is to one or more symptoms (for example, suppressing decalcification) for releasing osteoporosis disease, or is useful for the recalcification of inducing porotic bone (recalcification). Described peptide is useful as the preventive means of the paresthesia epilepsy of prevention of osteoporosis disease in mammal (patient who for example, has the osteoporosis risk).

We believe that similar mechanism is the reason of CAC, for example the aortic stenosis of calcification. Thereby in some embodiments, the present invention expection is used peptide described here or amino acid to suppressing or prophylactic symptom, and described disease is aortic stenosis, the osteoporosis of CAC, calcification for example, etc.

VI. other indication.

Be not limited to specific theory, we also believe peptide described here or amino acid pair, the various Other diseases of outbreak and/or one or more symptoms alleviate to(for) preventative or therapeutic ground are effective, described disease includes but not limited to, polymyalgia rheumatica (polymyalgia rheumatica), polyarteritis nodosa, scleroderma, lupus erythematosus, multiple sclerosis, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease (for example, asthma), Alzheimer's, AIDS and diabetes. Usually, described peptide is useful aspect relief of symptoms, and described symptom results from the inflammatory reaction of these diseases or relevant with the inflammatory reaction in these diseases.

VII. peptide/amino acid is to using.

Method of the present invention usually comprises to organism, preferably mammal, be more preferably the mankind, use one or more peptides of the present invention or amino acid to (or this peptide or the right analogies of amino acid). As described here, described peptide or amino acid are to can be according to any using in the multiple standards method, and described standard method includes but not limited to that (for example, nose spraying, oral cavity suck for injection, suppository, suction, etc.), the implantation of time controlled released, wear the paster of epidermis, etc. In particularly preferred embodiments, described peptide is Orally administered (for example, as syrup, capsule, powder, gelcap or tablet).

Described method can comprise uses single peptide or amino acid pair of planting of the present invention, or uses two or more different peptides or amino acid pair. Described peptide or amino acid is to can be used as monomer, or provides with form dimerization, oligomerization or poly. In some embodiments, the monomer that described polymer form can comprise connection (for example, connect by ionization or hydrophobic interaction), and some other polymer form can comprise the monomer (directly connect or connect via joint) that connects with covalent bond.

Although be of the present invention for making to describe in the mankind, it also is suitable for animal, for example, animal doctor's purposes. Thereby preferred organism is including, but not limited to the mankind, non-human primates, dog, horse, cat, pig, ungulate, largomorphs, etc.

Method of the present invention is not limited to and (has for example shown atherosclerotic one or more symptoms, hypertension, Mottling formation and breaking, the minimizing of for example heart attack of clinical events, angina pectoris or apoplexy, high-caliber plasma cholesterol, high-caliber low-density lipoprotein, high-caliber VLDL, or struvite albumen CRP for example, etc.) the mankind or non-human animal, also be useful in preventative environment. Thereby peptide of the present invention or amino acid can be applied to organism (or its analogies) and come the outbreak of one or more symptoms of one of atherosis and/or other indication described here of prevention of arterial/development. Here particularly preferred experimenter (has for example shown atherosclerotic one or more hazards, family history, hypertension, obesity, be addicted to drink, family history, high blood lipid, heart attack, angina or the apoplexy of blood LDL, VLDL, IDL or low HDL, diabetes or the diabetes of smoking, high blood cholesterol, high blood triglyceride, rising, etc.) and/or the experimenter of one of other situation described here.

In some embodiments, also can use peptide of the present invention and/or amino acid to before stimulating-formation and the circulation of β HDL like-particles, and/or promote reverse lipid transfer and detoxifcation.

Described peptide or amino acid also are useful to using for the associating inhibin, and this moment, they strengthened (for example, the collaborative enhancing) usually activity of the inhibin of application dosage, and/or allowed inhibin to use with lower dosage.

In addition, can use described peptide or amino acid to reducing or eliminate one or more symptoms of osteoporosis and/or diabetes and/or any other situation described here, and/or prevention/the suppress outbreak of one or more symptoms of osteoporosis and/or any other indication described here.

VIII. some preferred peptide and their preparation.

A) category-A Amphiphilic helix peptide.

Discovery of the present invention is that the peptide (" category-A peptide ") that comprises category-A Amphiphilic helix (amphipathic helix) can be alleviated atherosclerotic one or more symptoms. The feature of category-A peptide is the formation of alpha-helix, separating of its production polarity and non-polar residue, thereby form polarity and non-polar plane, the residue that has at the interface positively charged at polar-nonpolar, exist in the center of polar surface electronegative residue (referring to, for example, Anantharamaiah (1986) Meth.Enzymol, 128:626-668). Notice that the 4th extron of apo A-I produces category-A Amphiphilic helix structure when being folded into 3.667 residues/corner.

An especially effective category-A peptide, be called 18A (referring to, for example, Anantharamaiah (1986) Meth.Enzymol, 128:626-668) make amendment as described here, generation can be Orally administered and suppress or one or more symptoms that prevention of arterial is atherosis aspect the effective peptide of height. Be not limited to specific theory, it is believed that peptide of the present invention may work the oxidation of described seed Molecular remission LDL in vivo by collecting the seed molecule.

We have determined will improve in theory the lipid affinity in the quantity of the hydrophobic surface raising Phe of 18A residue, by Palgunachari et al. (1996) Arteriosclerosis, the computing method that Thrombosis ， ﹠Vascular Biology 16:328-338 describes is measured. In theory, the systematic residue of Phe is replaced and will be produced six kinds of peptides on the non-polar plane of 18A. Peptide with 2,3 and 4 Phe of increase will have respectively 13,14 and 15 units of theoretic lipid affinity (λ) value. Yet if the Phe that increases brings up to 5 from 4, λ value will rise to four units (to 19 λ units). Be increased to 6 or 7 Phe and will produce not too significantly raising (branch is clipped to 20 and 21 λ units). Therefore, we select the Phe (described peptide is called as 5F thus) of 5 increases. In a particularly preferred embodiment, described 5F peptide is closed, and wherein n terminal residue is acetylation and is amidated with carboxyl terminal residue.

New category-A peptide analog, 5F suppresses the pathological development in the atherosclerotic susceptible mouse. This new peptide analog 5F, use is according to the peptide dosage of the research (Levine et al. (1993) Proc.Natl.Acad.Sci.USA 90:12040-12044) of Levine etc., relatively suppresses effectiveness aspect the atherosclerotic of diet induced with mouse apoA-I (MoA-I) in these mouse.

Also produced many other the category-A peptide and alleviate shown aspect atherosclerotic one or more symptoms variation, but be the effectiveness of significance degree. Many this peptides in table 1, have been exemplified.

Table 1. is used for the illustrative analogies of the Amphiphilic helix of Apo A-I of the present invention.

¹Joint has added underscore.

NMA is N-methylamino o-tolyl.

Some preferred embodiment in, described peptide comprises the variant of 4F (the SEQ ID NO:8 of table 1) or D-4F, wherein one or two aspartic acid (D) is substituted by glutamic acid (E). Also be contemplated that peptide (for example, 4F or D-4F), wherein substituted by glutamic acid (E) from 1,2,3 or 4 amino acid of carboxyl terminal deletion and/or from carboxyl terminal 1,2,3 or 4 amino acid of deletion and/or one or two aspartic acid (D). In what peptide described here in office, can seal the N-end and use mantyl part (for example, N-methylamino o-tolyl) mark.

Although the various peptides of table 1 are with the aminoterminal acetyl group of protection or N-methylamino o-tolyl and protect the acid amides group of carboxyl terminal to illustrate that any one of these blocking groups can be eliminated or replace with another blocking group described here. In particularly preferred embodiments, described peptide comprises one or more D type amino acid described here. In some embodiments, each amino acid of the peptide of table 1 (for example, each enantiomer amino acid) is D type amino acid.

Be also noted that table 1 does not fully comprise. Use is in this instruction that provides, and can produce routinely other category-A Amphiphilic helix peptide that is fit to (for example, by conservative or semi-conservative property replacement (for example, D is substituted by E), interpolations, delete, etc.). Thereby for example, embodiment has utilized the body that blocks of any or multiple peptide (peptide of for example, being determined by SEQ ID Nos:5-23 and 42-in the table 1) in this demonstration. Thereby for example, SEQ ID NO:24 has illustrated 14 amino acid whose peptides that comprise from the C-end of 18A, and described 18A comprises one or more D amino acid, and SEQ ID NOS:25-41 has illustrated that other blocks body.

Longer peptide also is fit to. This longer peptide may fully form the category-A Amphiphilic helix, and perhaps the category-A Amphiphilic helix can form one or more domains of described peptide. In addition, the present invention expects the polymeric pattern of described peptide. Thereby, for example, can be coupled at together at the peptide of this explanation (directly or via having the amino acid whose joint of one or more insertions (for example, carbon joint, or one or more amino acid)). Illustrative pdef polypeptide comprises the peptide of 18A-Pro-18A and SEQ ID NO:81-88; comprise in some embodiments one or more D amino acid; preferred each amino acid is such as D amino acid described here, and/or it is protected to have one or two end.

B) has other category-A Amphiphilic helix peptide mimics aromatic series or aliphatic residue, apoA-I at non-polar plane.

In some embodiments, the present invention also provides the category-A amphiphilic helical peptides of modifying.Some preferred peptide has mixed one or more aromatic residues, for example 3F at the center of non-polar plane ^{C π}(as in 4F, existing), or mix one or more aliphatic residues, for example 3F at the center of non-polar plane ^{I π}Be not limited to specific theory, we believe at peptide 3F ^{C π}Non-polar plane on central aromatic residue; because existence at the center of non-polar plane p electronics; allow the hydrophobic lipid alkyl chain of hydrone infiltration near peptide-lipid complex; it allows entering of reactive oxygen class (for example lipid peroxide) conversely, protects them to avoid cell surface.Similarly, we believe that also the center at non-polar plane has the peptide of aliphatic residue, for example, and 3F ^{I π}, will be similarly but resemble 3F by halves ^{C π}Equally work effectively.

Preferred Toplink will be urged struvite HDL and change into anti-inflammatory HDL, or make anti-inflammatory HDL anti-inflammatory stronger, and/or reduce the inductive monocyte chemotactic activity of LDL that arterial wall cell produces, and described effect is equal to or greater than other peptide shown in D4F or the table 1.Shown that this active peptide is improving atherosclerosis and other struvite situation, for example, rheumatoid arthritis, lupus erythematosus, polyarteritis nodosa, osteoporosis, Alzheimer, congestive heart failure, endothelial function disturbance and virus disease be A type influenza for example, for example the disease aspect of multiple sclerosis is useful.

The example of some preferred peptide of table 2..

C) littler peptide.

Also have a surprising discovery to be, some little peptide of forming by minimum three aminoacid, preferably (but dispensable) one or more described aminoacid are amino acid whose D-stereoisomers, with have hydrophobic domains and interact to allow lipid protein, with the hydrophilic-structure territory to allow water solublity to a certain degree, also have significant antiinflammatory property.Be not limited to specific theory, it is believed that for example Ox-PAPC, POVPC, PGPC and PEIPC combine for the described peptide and the formation of the phospholipid of short struvite oxidation required " seed molecule ".Because many struvite situations are at least in part by the mediation of the lipid of oxidation, we believe, described peptide of the present invention is being effectively improving aspect the situation, the formation of the known or lipid that results from the biologic activity oxidation under a cloud of described situation.These are including, but not limited to atherosclerosis, rheumatoid arthritis, lupus erythematosus, polyarteritis nodosa, pulmonary disease, asthma, multiple sclerosis, Alzheimer, diabetes and osteoporosis.Described " little peptide " usually length range from 3 aminoacid to about 15 aminoacid, preferred from about 4 aminoacid to about 10 or 11 aminoacid, most preferred from about 4 to about 8 or 10 aminoacid.Described peptide is usually to have hydrophobic end amino acid or to have been handled end amino acid by hydrophobization be feature by adhering to one or more hydrophobic " protection " group.

In some embodiments, peptide can be being feature as shown in the formula I:

X ¹-X ²-X ³ _n-X ⁴ I

Wherein, n is 0 or 1, X ¹Be hydrophobic amino acid and/or have hydrophobic blocking group, X ⁴Be hydrophobic amino acid and/or have hydrophobic blocking group; With when n is 0, X ²Be acidity or basic amino acid; When n is 1: X ²And X ³Be acidic amino acid, basic amino acid, aliphatic amino acid or aromatic amino acid independently, work as X like this ²When being acidic amino acid; X ³Be basic amino acid, aliphatic amino acid or aromatic amino acid; Work as X ²When being basic amino acid; X ³Be acidic amino acid, aliphatic amino acid or aromatic amino acid; With work as X ²When being aliphatic or aromatic amino acid, X ³Be acidic amino acid or basic amino acid.

Longer peptide (for example, reaching 10,11 or 15 aminoacid) also is to expect in the scope of the present invention.Usually, wherein said shorter peptide (for example, peptide according to formula I) feature is tart, alkaline, aliphatic or aromatic amino acid, and the feature of described longer peptide is to comprise two or more such amino acid whose, tart, alkaline, aliphatic or aromatic domains.

1) functional characteristic of active small peptide

A surprising discovery of the present invention is, many physical propertys (are for example predicted little peptide of the present invention, be less than 10 aminoacid, preferably be less than 8 aminoacid, preferred from about 3 to about 5 or 6 aminoacid) make the stronger and releasing mammal of HDL anti-inflammatory atherosclerosis and/or be other pathological changes of feature with the inflammatory reaction.Described physical property is included in highly dissoluble (for example, greater than about 4mg/mL) and the dissolubility in the aqueous buffer solution of pH 7.0 in the ethyl acetate.At contact phospholipid, for example 1, during two myristoyls of 2--sn-glyceryl-3-phosphocholine (DMPC), in aqueous environments, effective especially little inducing peptide or participate in the about 7.5nm of the diameter (formation of ± 0.1nm) particle, and/or induce or participate in the formation of stacked bilayer, bilayer size general 3.4 to 4.1nm, the about 2nm in gap between the bilayer in stacked, and/or also induce or participate in the formation of the loose structure of about 38nm.Some preferred embodiment in, described little peptide has the molecular weight that is lower than about 900Da.

Thereby, in some embodiments, the present invention expects the little peptide of one or more symptoms of improving struvite situation, wherein said peptide: length range from about 3 to about 8 aminoacid, preferably from about 3 to about 6 or 7 aminoacid, with preferred from about 3 to about 5 aminoacid, can be dissolved in ethyl acetate with concentration greater than about 4mg/mL; Dissolve in the aqueous buffer solution of pH 7.0; When aqueous environments contacts with phospholipid, form the particle of the about 7.8nm of diameter, and/or form stacked bilayer, bilayer size general (on the order of) 3.4 to 4.1nm, the about 2nm in interval between the stacked middle bilayer; Have and be lower than about 900 daltonian molecular weight; To urge struvite HDL and be converted into anti-inflammatory HDL, or make anti-inflammatory HDL anti-inflammatory stronger; Do not have aminoacid sequence Lys-Arg-Asp-Ser (SEQ ID NO:238), wherein Lys-Arg-Asp and Ser are L aminoacid.In some embodiments, these little peptide protection phospholipid are to the oxidation of antioxidant.

Though these little peptides needn't be limited like this, in some embodiments, these little peptides can comprise little peptide as described below.

2) tripeptides.

Have been found that, can synthesize some tripeptides (3 amino acid whose peptides), its character that has shown expectation described here (for example, short struvite HDL is changed into the ability of anti-inflammatory HDL, reduce the active ability of the inductive monocyte chemotactic of LDL that arterial wall cell produces, improve the ability of preceding-β HDL, or the like).In some embodiments, the feature of described peptide is formula I, and wherein N is zero, the following formula II that is shown as:

X ¹-X ²-X ⁴ II

End amino acid (X wherein ¹And X ⁴) be hydrophobic, be because of hydrophobic side chain, or because side chain or C and/or N-terminal (are for example sealed by one or more hydrophobic blocking groups; the N-end by Boc-, Fmoc-, nicotinoyl (Nicotinyl)-; or the like sealing, C-is terminal by (tBu)-OtBu, or the like sealing).In some embodiments, X ²Aminoacid is tart (for example, aspartic acid, glutamic acid, or the like) or alkalescence (for example, histidine, arginine, lysine, or the like).Described peptide can all be a L aminoacid, or comprises one or more or whole D aminoacid.

Some preferred tripeptides of the present invention is including, but not limited to the peptide shown in the table 3.

Table 3. has hydrophobic blocking group and example acid, amino acid whose some the preferred tripeptides of alkalescence or histidine central authorities.

Though the peptide of table 3 is to illustrate with specific blocking group, notice that these groups can be replaced by other blocking group described here, and/or one or more blocking group that illustrates can be removed.

3) the little peptide of the acid and basic amino acid of central authorities is arranged.

In some embodiments, peptide of the present invention from four aminoacid to about ten aminoacid.End amino acid usually is hydrophobic, is because of hydrophobic side chain, or because end amino acid has one or more hydrophobic blocking groups, end amino acid (X ¹And X ⁴) be hydrophobic, or because hydrophobic side chain, or because side chain or C and/or N-terminal by one or more hydrophobic blocking groups sealings (for example, the N-end by Boc-, Fmoc-, nicotinoyl-, or the like sealing, C-is terminal by (tBu)-OtBu, or the like sealing).Usually, the middle body of described peptide comprises basic amino acid and acidic amino acid (for example, in 4 bodies), or comprises basic domain and/or acid domain in long molecule.

These limbs can be by formula I representative, wherein X ¹And X ⁴Be hydrophobic and/or have hydrophobic blocking group described here, work as X ³Be the time X of alkalescence ²Be tart, or work as X ³X when being tart ²Be alkaline.Described peptide can all be a L aminoacid, or comprises one or more or whole D aminoacid.

Some preferred peptide of the present invention is including, but not limited to the peptide shown in the table 4.

Table 4. has the illustrative example of the little peptide of central authorities' acidity and basic amino acid.

Though the peptide of table 4 is to illustrate with specific blocking group, notice that these groups can be replaced by other blocking group described here, and/or one or more blocking group that illustrates can be removed.

4) has the little peptide that acidity or basic amino acid also have central aliphatic amino acid simultaneously in central authorities.

In some embodiments, peptide of the present invention from four aminoacid to about ten aminoacid.End amino acid usually is hydrophobic, is because of hydrophobic side chain, or because end amino acid has one or more hydrophobic blocking groups.End amino acid (X wherein ¹And X ⁴) be hydrophobic, be because hydrophobic side chain, or because side chain or C and/or N-terminal by one or more hydrophobic blocking groups sealings (for example, the N-end by Boc-, Fmoc-, nicotinoyl-, or the like sealing, C-is terminal by (tBu)-OtBu, or the like sealing).Usually, the middle body of described peptide comprises alkalescence or acidic amino acid and aliphatic amino acid (for example, in 4 bodies), or comprises basic domain or acid domain and aliphatic structure territory in long molecule.

These limbs can be by formula I representative, wherein X ¹And X ⁴Be hydrophobic and/or have hydrophobic blocking group described here, work as X ³X when being aliphatic ²Be acid or alkaline, or work as X ³Be acid or alkaline time X ²Be aliphatic.Described peptide can all be a L aminoacid, or comprises one or a plurality of or whole D aminoacid.

Some preferred peptide of the present invention is including, but not limited to the peptide shown in the table 5.

Table 5. has the example that acidity or basic amino acid also have some preferred peptide of central aliphatic amino acid simultaneously in central authorities.

Though the peptide of table 5 is to illustrate with specific blocking group, notice that these groups can be replaced by other blocking group described here, and/or one or more blocking group that illustrates can be removed.

5) has the little peptide that acidity or basic amino acid also have central aromatic amino acid simultaneously in central authorities.

In some embodiments, peptide of the present invention from four aminoacid to about ten aminoacid.End amino acid usually is hydrophobic, is because of hydrophobic side chain, or because end amino acid has one or more hydrophobic blocking groups, end amino acid (X ¹And X ⁴) be hydrophobic, be because hydrophobic side chain, or because side chain or C and/or N-terminal by one or more hydrophobic blocking groups sealings (for example, the N-end by Boc-, Fmoc-, nicotinoyl-, or the like sealing, C-is terminal by (tBu)-OtBu, or the like sealing).Usually, the middle body of described peptide comprises alkalescence or acidic amino acid and aromatic amino acid (for example, in 4 bodies), or comprises basic domain or acid domain and aromatic structure territory in long molecule.

These limbs can be by formula I representative, wherein X ¹And X ⁴Be hydrophobic and/or have hydrophobic blocking group described here, work as X ³X when being aromatic ²Be acid or alkaline, or work as X ³Be acid or alkaline time X ²Be aromatic.Described peptide can all be a L aminoacid, or comprises one or a plurality of or whole D aminoacid.Five body constituents can be by the small modified forms representative of formula I, wherein X ⁵As shown in table 6 is inserted into, and X wherein ⁵It usually is aromatic amino acid.

Some preferred peptide of the present invention is including, but not limited to the peptide shown in the table 6.

Table 6. has the example that acidity or basic amino acid also have some preferred peptide of central aromatic amino acid simultaneously in central authorities.

Though the peptide of table 6 is to illustrate with specific blocking group, notice that these groups can be replaced by other blocking group described here, and/or one or more blocking group that illustrates can be removed.

6) has aromatic amino acid in the center or a little peptide of the aromatic amino acid separated by histidine.

In some embodiments, peptide of the present invention is characterised in that the p electronics of the central authorities that are exposed to molecule, it allows the hydration of particle, this allows the lipid of the short struvite oxidation of described peptide particle trapping, for example fatty acid peroxidase thing and contain the phospholipid of arachidonic oxidation product in the sn-2 position.

In some embodiments; these peptides are made up of minimum 4 aminoacid and maximum about 10 aminoacid; preferentially (rather than necessarily) one or more aminoacid are arranged is amino acid whose D-stereoisomers; end amino acid is hydrophobic; be because hydrophobic side chain, or because described end amino acid have one or more hydrophobic blocking groups (for example, the N-end by Boc-, Fmoc-, nicotinoyl-; or the like sealing and C-terminal by sealings such as (tBu)-OtBu groups).Be not to have acidity or basic amino acid in central authorities, these peptides usually have aromatic amino acid in central authorities, or have the aromatic amino acid of being separated by histidine in the central authorities of described peptide.

Some preferred peptide of the present invention is including, but not limited to the peptide shown in the table 7.

Table 7. has aromatic amino acid or the example of the peptide in the aromatic amino acid separated by one or more histidine or aromatic structure territory in the center.

Though the peptide of table 7 is to illustrate with specific blocking group, notice that these groups can be replaced by other blocking group described here, and/or one or more blocking group that illustrates can be removed.

7) general introduction of tripeptides and tetrapeptide.

For the sake of clarity, many tripeptides of the present invention and tetrapeptide are following makes overview at table 8.

The general structure of table 8. some peptide of the present invention.

When expecting long peptide, X ²And X ³Can representative structure territory (for example, the two or more amino acid whose zone of specified type) rather than independent aminoacid.Table 8. is illustrative and not restrictive.Use can easily be identified the peptide that other is fit in this instruction that provides.

8) paired aminoacid and dipeptides.

The present invention relates to such discovery, some aminoacid is right, and co-administered mutually or connection forms the dipeptides with one or more character described here.Thereby, be not limited to specific theory, it is believed that when described aminoacid to mutual when co-administered, as described here, they can participate in or induce the formation of micelle (micelles) in vivo.

Be similar to other little peptide described here, it is believed that peptide comprises highly dissoluble (for example, greater than about 4mg/mL), the deliquescent physical property in the aqueous buffer solution of pH 7.0 in the ethyl acetate to associating in vivo, and showing.At contact phospholipid, for example 1, during two myristoyls of 2--sn-glyceryl-3-phosphocholine (DMPC), in aqueous environments, it is believed that described aminoacid is to inducing or participated in diameter about 7.5nm (formation of ± 0.1nm) particle, and/or induce or participated in the formation of stacked bilayer, bilayer size general 3.4 is to 4.1nm, the about 2nm in the gap between the bilayer in stacked, and/or also induce or participate in the formation of the loose structure of about 38nm.

In addition, believe further that described aminoacid is gone up corresponding character to showing the physiology below one or more:

1. they will be urged struvite HDL and change into anti-inflammatory HDL or make anti-inflammatory HDL anti-inflammatory stronger;

2. they reduce the inductive monocyte chemotactic activity of LDL that arterial wall cell produces;

3. they stimulate formation and the circulation of preceding-β HDL;

4. they improve the HDL cholesterol; And/or

5. they improve HDL paraoxonase activity.

Described aminoacid is used (order ground or side by side use to can be used as independently aminoacid, for example, in the preparation of combination), or they can be directly or via joint (for example, the joint that PEG joint, carbon joint, ramose joint, straight chain joint, heterocyclic joint, deutero-lipid form, or the like) by covalently coupling.In some embodiments, described aminoacid is to covalently connecting via peptide bond to form dipeptides.In various embodiments, though described dipeptides usually comprises two aminoacid of blocking group with the connection separately, the present invention also comprises such dipeptides, and wherein only an aminoacid has one or more blocking groups.

Described aminoacid is to usually comprising aminoacid, and wherein each aminoacid is connected at least one blocking group (for example, hydrophobic blocking group described here).Described aminoacid can be D or L type.In some embodiments, wherein said aminoacid comprises not to be had interconnective rightly, and each aminoacid has two blocking groups (for example, as the molecule 1 in the table 9 and 2).

The illustrative aminoacid of the present invention of table 9. is right

^*This is usually united second aminoacid and uses.

^*In some embodiments, these dipeptides are co-administered mutually.

^{* *}In some embodiments, this peptide individually or with one of other peptide described here combined administration.

By provide shielded aminoacid to and/or dipeptides, then according to one or more physics as mentioned above and/or physiological character screen described aminoacid right/dipeptides, it is right easily to identify suitable aminoacid.In some embodiments, the present invention got rid of the aminoacid that comprises aspartic acid and phenylalanine to and/or dipeptides.In some embodiments, the present invention got rid of such aminoacid to and/or dipeptides, one of them aminoacid is (-)-N-[(trans-4-isopropyl cyclohexane extraction) carbonyl]-D-phenylalanine (nateglinide).

In some embodiments, described aminoacid comprises and by acidic amino acid (for example is independently selected from, aspartic acid, glutamic acid, or the like), basic amino acid (for example, lysine, arginine, histidine, or the like) and the group that constitutes of nonpolar amino acid (for example, alanine, valine, leucine, isoleucine, proline, phenylalanine, tryptophan, methionine, or the like) is right.In some embodiments, wherein said first aminoacid is acid or alkaline, and second aminoacid is that nonpolar and wherein said second aminoacid is acid or alkaline, and first aminoacid is nonpolar.In some embodiments, wherein first aminoacid is tart, and second aminoacid is alkaline, vice versa (referring to, for example, table 10).

Similarly combination can by use dipeptides to obtaining.Thereby for example in some embodiments, the molecule 3 and 4 in the table 9 is co-administered mutually.

Some common peptide of table 10. is right.

To notice, these aminoacid are right/and dipeptides is illustrative and not restrictive.Use is in this instruction that provides, can easily determine other aminoacid that is fit to right/dipeptides.

D) other peptide is modified body.

Surprising discovery is, peptide described here, particularly when their fusion during one or more D aminoacid, kept the active of them and also can use orally.In addition, this Orally administered generation is relatively effectively absorbed and the significant serum half-life, alleviates atherosclerosis or is the effective ways of one or more symptoms of other pathological changes of feature with the inflammatory process thereby provide.

Use is in this instruction that provides, and those of skill in the art can modify the peptide that is exemplified routinely and produce other similar peptide of the present invention.For example, can carry out conventional conservative or half conservative to existing aminoacid and replace (for example, E replaces D).Can use Palgunachari et al. (1996) Arteriosclerosis, Thrombosis ， ﹠amp; The computational methods that Vascular Biology 16:328-338 describes are predicted the influence of various replacements for the lipid affinity of the peptide that produces.As long as kept α-Luo Xuanjiegou, described peptide can be extended or shorten.In addition, can replace so that the peptide that produces is similar to the peptide that experimenter's species endogenous produces more.

In some embodiments, peptide of the present invention is included in United States Patent (USP) 4,643, and the peptide of " D " type of describing in 988 more preferably has " D " type with link coupled one or two end of blocking group.In some embodiments, at least 50% enantiomer aminoacid is " D " type, more preferably at least 80% enantiomer aminoacid be " D " type and most preferably at least 90% so that all enantiomer aminoacid be " D " type aminoacid.

Though, in some embodiments, peptide of the present invention has utilized the aminoacid of natural generation or the amino acid whose D type of natural generation, the aminoacid that takes place with non-natural (for example, methionine sulfoxide, methionine methyl sulfonium, nor-leucine, episilon amino caproic acid, 4-aminobutyric acid, four water isoquinolin-3-carboxylic acid, 8-aminocaprylic acid, 4-aminobutyric acid, Lys (N (ε)-trifluoroacetyl), α-An Jiyidingsuan, or the like) replacement also expect.

Except describing the peptide, intend peptide (peptidomimetics) at this and also expect at this.Peptide analogues is generally used for pharmaceuticals industry, as the non-peptide medicine with the character that is similar to the template peptide.The non-peptide compound of these types is called as " peptide mimics (peptide mimetics) " or " plan peptide " (Fauchere (1986) Adv.Drug Res.15:29; Veber and Freidinger (1985) TINSp.392; With Evans et al. (1987) J.Med.Chem.30:1229), develop by means of computerized molecule modeling usually.Be similar to the peptide mimics for the treatment of upward useful peptide on the structure and can be used to produce equal treatment or preventive effect.

Usually, intend peptide and structurally be similar to example polypeptide (for example, 4F described here, SEQ ID NO:258), but the randomly selected freedom-CH of one or more peptide bonds is arranged ₂NH-,-CH ₂S-,-CH ₂-CH ₂-,-CH=CH-(cis and trans) ,-COCH ₂-,-CH (OH) CH ₂-,-CH ₂The key of the group that SO-etc. constitute substitutes, by methods known in the art with below with reference to the method that further describes in the document: Spatola (1983) Chemistry and Biochemistry of Amino Acids p.267in, Peptides, and Proteins, B.Weinstein, eds., Marcel Dekker, New York; Spatola (1983) Vega Data 1 (3) Peptide Backbone Modifications. (general review); Morley (1980) Trends Pharm Sci pp.463-468 (general review); Hudson et al. (1979) Int J Pept Prot Res 14:177-185 (CH ₂NH-, CH ₂CH ₂-); Spatola et al. (1986) Life Sci 38:1243-1249 (CH ₂-S); Hann, (1982) J Chem Soc Perkin TransI 307-314 (CH-CH-, cis and trans); Almquist et al. (1980) J Med Chem.23:1392-1398 (COCH ₂-); Jennings-White et al. (1982) Tetrahedron Lett.23:2533 (COCH ₂-); Szelke, M.et al., European Appln.EP 45665 (1982) CA:97:39405 (1982) (CH (OH) CH ₂-); Holladay et al. (1983) TetrahedronLett 24:4401-4404 (C (OH) CH ₂-); And Hruby (1982) Life Sci., 31:189-199 (CH ₂-S-)).

Particularly preferred non-peptide bond is-CH ₂NH-.This peptide mimics may have significant preferred than polypeptide embodiment, comprises, for example: the antigenicity of more economical production, higher chemical stability, enhanced pharmacological property (half-life, absorption, effectiveness, effect, or the like), reduction, or the like.

In addition, can pass through the known method in this area (Rizo and Gierasch (1992) Ann.Rev.Biochem.61:387), for example, can form the inside cysteine residues that intramolecular disulfide bond makes the peptide cyclisation by adding, to produce and comprise consensus sequence or the identical in fact annular arrangement consensus sequence variant, peptide described here or the peptide (peptide that comprises cyclisation) of constraint.

IX. the Function detection of peptide

Some peptide of the present invention this be by various chemical formulas (for example, formula I, as above) and/or describe by particular sequence.Yet in some embodiments, the characteristics of preferred peptide of the present invention are the functional character that one or more are following:

1. they will be urged struvite HDL and change into anti-inflammatory HDL or make anti-inflammatory HDL anti-inflammatory stronger;

2. they reduce the inductive monocyte chemotactic activity of LDL that arterial wall cell produces;

3. they stimulate formation and the circulation of preceding-β HDL;

4. they improve the HDL cholesterol; And/or

5. they improve HDL paraoxonase activity.

Particular peptide disclosed herein, and/or with the corresponding peptide of various chemical formulas described here, can easily test one or more these activity on demand.

Each screening technique according to these functional characters is known for those skilled in the art.In addition, at this such screening has been described in an embodiment.Especially, notice, in PCT/US01/26497 (WO 02/15923), illustrated monocyte chemotactic activity, HDL cholesterol and the active detection of HDL HDL paraoxonase.The detection that is used for determining the character of the struvite and/or anti-inflammatory of HDL is carried out as described below.

A) measure HDL struvite/the anti-inflammatory characteristic

1) monocyte chemotactic activity (MCA) detects

Preparation lipoprotein, human artery's wall coculture and mononuclear cell are as (VanLenten et al. (2002) Circulation, 106:1127-1132) the mensuration monocyte chemotactic activity (MCA) of previous description.Under the situation of the HDL that does not have or exist the experimenter, bioassay standard contrast LDL inducing to MCA.Numerical value under the situation that does not have HDL is standardized as 1.0.Numerical value greater than 1.0 after adding HDL shows short struvite HDL; Be lower than 1.0 numerical value and show anti-inflammatory HDL.

2) acellular detection

Described acellular detection is the modification of previous disclosed use PEIPC as fluorescence induction compositions and methods 9.Briefly, separate HDL by the dextran sulfate method.The Sigma " HDL cholesterol reagent " (Catalog No.352-3) that will contain dextran sulfate and magnesium ion is dissolved in (10.0mg/mL) in the distilled water.The dextran sulfate solution and the 500 μ l test blood plasma of 50 microlitres is mixed, at room temperature hatched 5 minutes, subsequently 8, centrifugal 10 minutes of 000g.(Arlington TX) measures after the cholesterol for Cat.No.2340-200, Thermo DMA Company, and the supernatant that contains HDL is used in the experiment using the cholesterin detection reagent box.We have reported previously (Navabet al. (2001) J Lipid Res 1308-1317), and compares by the isolating HDL of conventional ultracentrifugation, isolating by this method HDL with the passivation of biological activity phospholipid to similar degree.In order to measure character from the struvite/anti-inflammatory of the HDL sample of patient and contrast, use the variation of fluorescence intensity aspect, described variation is not exist or exists under the situation of testing HDL as DCFH by the result of PEIPC oxidation.DCFH-DA is dissolved in the fresh methanol with 2.0mg/mL, at room temperature hatches and kept in Dark Place 30 minutes, produce the release of DCFH.Revise described detection be fit to with a large amount of sample of plate reader analysis.For this reason, utilize polystyrene microtiter plates flat, black (Microfluor2, Cat.No.14-245-176, Fisher).Be distributed in the microtitration plate HDL that contains the dextran sulfate supernatant (final concentration 10 μ g/ml cholesterol) of 10 μ l PEIPC solution (final concentration 50 μ g/ml) and 90 μ l also mixed.Then flat board was hatched on rotator 1.0 hours at 37 ℃.Add 10 μ l DCFH solution (0.2mg/mL) to each reacting hole then, mix, and hatched extra 2 hours 37 ℃ of rotations.Subsequently, in excitation wavelength and the emission wavelength of 530nm and the blocking of 515nm of 485nm, photomultiplier sensitivity be set to " in ", with plate reader (Spectra Max, Gemini XS; Molecular Devices) measures fluorescence.The numerical value of intra-and interassay variability is respectively 5.3 ± 1.7% and 7.1 ± 3.2%.Numerical value under the situation that does not have HDL is normalized to 1.0.Numerical value greater than 1.0 after adding test HDL shows short struvite HDL; Be lower than 1.0 numerical value and show anti-inflammatory HDL.

3) other step

By previous disclosed method (Scheidt-Nave et al. (2001) J Clin EndocrinolMetab., 86:2032-2042; Piguet et al. (1987) J Experiment Med., 166, the 1280-1289) blood plasma level of mensuration interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α).(Navab et al. (1997) J Clin Invest according to previous description, 99:2005-2019) use test kit (Sigma) to measure total plasma cholesterol, triglyceride, LDL-cholesterol, HDL cholesterol and glucose, use is from Immunodiagnostik (ALPCO Diagnostics, Windham, NH) sandwich EIA enzyme immunoassay measure the hs-CRP level (Rifai et al. (1999) Clin Chem., 45:2136-2141).Determine statistical significance with model I ANOVA, significantly be defined as the value of p＜0.05.

4) physical property of the little peptide of screening

A surprising discovery of the present invention is, many physical propertys (are for example predicted little peptide of the present invention, be less than 10 aminoacid, preferably be less than 8 aminoacid, preferred from about 3 to about 5 or 6 aminoacid) make the stronger and releasing mammal of HDL anti-inflammatory atherosclerosis and/or be other pathological changes of feature with the inflammatory reaction.As described in this, described physical property is included in highly dissoluble (for example, greater than about 4mg/mL) and the dissolubility in the aqueous buffer solution of pH 7.0 in the ethyl acetate.At contact phospholipid, for example 1, during two myristoyls of 2--sn-glyceryl-3-phosphocholine (DMPC), in aqueous environments, effective especially little peptide forms the about 7.5nm of diameter (± 0.1nm) particle, and/or form stacked bilayer, bilayer size general 3.4 to 4.1nm, the about 2nm in gap between the bilayer in stacked, and/or also form the loose structure of about 38nm.Some preferred embodiment in, described little peptide has the molecular weight that is lower than about 900Da.

In fact can easily screen any little peptide according to one or more these character, for example, as what in this embodiment 3, describe.In fact, contain more than about 10 ⁴, or 10 ⁵, preferred more than about 10 ⁶Or 10 ⁷And it is most preferred more than about 10 ⁸Or 10 ⁹Plant the little peptide combinatorial library of little peptide, can use the method for well known to a person skilled in the art to produce.Described peptide library can be a random library, or alternatively, in some embodiments, the little peptide of making according to the one or more chemical formulas that provide at this is provided in described library.

One or more physical propertys as described above for example, use the high throughput screening method easily to screen described peptide library then.The positive peptide of test has probably and makes that the HDL anti-inflammatory is stronger and alleviate atherosclerosis and/or be the ability of other pathological changes of feature with the inflammatory reaction in mammal in these are analyzed.

Notice that above-mentioned screening technique only is illustrative, rather than restrictive.Use can easily be provided for other detection of the desired function character of peptide in this instruction that provides.

X. peptide preparation.

A) general synthetic method

It is chemically synthetic that the peptide that uses among the present invention can use the chemical peptide synthetic technology of standard, or when described peptide did not comprise " D " amino acid residue, described peptide can be easily by recombinant expressed especially.When recombinant expressed " D " polypeptide, host organisms (for example, antibacterial, plant, fungal cell, or the like) can be cultured in a kind of environment, wherein ad hoc provide one or more aminoacid to described organism with the D type.Recombinant expressed peptide has mixed those D aminoacid in this system then.

In some embodiments, can use the aminoacyl-tRNA synzyme of the amino acid whose modification of identification D that D aminoacid is incorporated in the recombinant expressed peptide.

Some preferred embodiment in, described peptide is any chemically synthetic by in the peptide synthetic technology of multiple liquid phase well known by persons skilled in the art or solid phase.Solid phase synthesis is the preferable methods of chemosynthesis polypeptide of the present invention, and in solid phase synthesis, the C-end amino acid of sequence is attached on the insoluble holder, the aminoacid of remainder in the successive subsequently interpolation sequence.The technology of solid phase synthesis is well known to a person skilled in the art, for example, and by Barany and Merrifield (1963) Solid-Phase Peptide Synthesis; Pp.3-284 in The Peptides:Analysis, Synthesis, Biology.Vol.2:Special Methods in PeptideSynthesis, Part A.; Merrifield et al. (1963) J Am.Chem.Soc., 85:2149-2156, and Stewart et al. (1984) Solid PhasePeptide Synthesis, 2nd ed.Pierce Chem.Co., Rockford, Ill has described.

In one embodiment, described peptide is to use benzhyderylamine resin (Beckman Bioproducts, the NH of 0.59mmol ₂/ g resin) synthetic as solid support by the solid-phase peptide synthesis step.COOH end amino acid (for example, t-butyl carbonyl-Phe) be attached on the solid support by 4-(oxygen methyl) phenylacetyl group.This is the key more stable than the benzyl ester bond of routine, and the peptide after finishing still can come cracking by hydrogenization.Use formic acid to be used to this purpose as the transfer hydrogenation effect of hydrogen donor.Be used for the detailed protocol of the analysis of the synthetic and synthetic peptide of peptide, following Anantharamaiah et al. (1985) J.Biol.Chem., 260 (16): described in the supplementary issue of reprinting books in a reduce format of 10248-10255.

Notice, at peptide, particularly comprise in the chemosynthesis of the amino acid whose peptide of D, except the full length product of expectation, syntheticly produce many peptides that block usually.Purge process (for example, HPLC) generally causes a large amount of losses of full length product.

Discovery of the present invention is, particularly (for example, in the middle of D-4) synthetic, for fear of the loss during a kind of peptide of microscler formula of purification, can dialyse and use mixture, thereby cancel last HPLC purification at the D peptide.This mixture has lost about 50% effectiveness (for example, every part of weight of protein product) of highly purified product, but described mixture contains the peptide of having an appointment more than 6 times, thereby gross activity is higher.

B) mix D type aminoacid.

Can be by simply in chemosynthesis, using the deutero-amino acid residue of D-type, D-aminoacid is incorporated into one or more positions in the peptide.Be used for the synthetic D type of solid-phase peptide residue can obtain from many suppliers commerciality (referring to, for example, Advanced Chem Tech, Louisville; Nova Biochem, San Diego; Sigma, St Louis; Bachem California Inc., Torrance, etc.).D-type aminoacid can fully be omitted, or is incorporated into any position in the peptide by expectation.Thereby, for example, in some embodiments, described peptide can comprise single D-aminoacid, and in other embodiments, described peptide comprises at least two, usually at least three, more generally at least four, the most at least five, preferably at least six, preferred at least seven and most preferred at least eight D aminoacid.In particularly preferred embodiments, be D type aminoacid every (enantiomer) aminoacid basically.In some embodiments, at least 90%, preferred at least 90%, preferred at least 95% enantiomer aminoacid is D-type aminoacid.In a particularly preferred embodiment, be exactly D-type aminoacid every (enantiomer) aminoacid basically.

C) liquid-phase synthesis process

In some embodiments, peptide of the present invention can use liquid phase process easily synthetic.A kind of such synthetic schemes has been described in attached Fig. 1 and 2.

In this scheme, the aminoacid in the peptide of A, B and C representative expectation.X-represents permanent alpha-amido blocking group.Y represents permanent α-carboxy protective group.If N-and C-end have side chain functionalities, alphabetical m and n represent the side chain protected group.Side chain protected group o and p are blocking groups, and described blocking group can be by for example using catalytic transfer hydrogenation effect (Anantharamaiah and Sivanandaiah (1977) the Chem Soc.Perkin Trans.490:1-5 of ammonium formate as hydrogen donor under (neutrality) condition; With Babiker et al. (1978) J.Org.Chem.44:3442-3444) processing be removed, wherein side chain protected group m and p and alpha-amido and α-carboxy protective group are stable.HOBT-HBTU represents condensation reagent, observes minimum racemization under this.

Have I-hydroxybenzotriazole-2 (H-benzotriazole-1-yl)-1,1,3,3-tetramethylammonium hexafluorophosphoric acid ester (HOBT-HBTU) and a small amount of tertiary amine for example under the situation of diisopropylethylamine (DIEA), add 2 normal H among the activated amino acid X-A (m) in DMF ₂N-B (n)-COO ^-DIEA salt, and at room temperature stir and spend the night.Use the acid of excess of ammonia base for activatory carboxylic acid, homologation reaction proceeds to be finished, and wherein alpha-amido is free, and carboxyl is a DIEA salt by temporary protection.Make use citric acid (10%) with the reactant mixture acidify, use ethyl acetate extraction.In this process, free amino acid is stayed in the citric acid.After the water flushing ethyl acetate, extract the dipeptides free acid of the terminal protection of N-with 5% sodium bicarbonate solution, and acidify.With ethyl acetate extraction dipeptides free acid, with organic layer drying (Na ₂SO ₄), evaporating solvent obtains described dipeptides free acid.With the aminoacid and the described dipeptides free acid reaction acquisition tripeptides of suitably protection, wherein alpha-amido is free by in a similar manner, and carboxyl is a DIEA salt by temporary protection.In order to obtain tetrapeptide, use the suitably aminoacid of carboxy protective of HOBT-HBTU condensation.Because final tetrapeptide is protected peptide, the reactant mixture in ethyl acetate after the extraction condensation, the careful washing of use heavy carbonate (5%) and citric acid (5%) washes with water then.These washing process will be removed excessive free acid and free alkali and condensation reagent.Use ethyl acetate (or ether) and the protected peptide of petroleum ether precipitation then.Under the situation that has freshly prepd palladium black (Pd is black), use ammonium formate then, make the effect of protected free peptide experience catalytic transfer hydrogenation as hydrogen donor.This reaction can almost carried out under the neutrallty condition, thereby does not influence the side chain protected group of acid-sensitive sense.This process will be removed the blocking group on aminoacid B and the C.The embodiment of this step provides hereinafter, uses synthesizing of SEQ ID NO:256.

Notice that this reaction scheme is illustrative and not restrictive.Use is in this instruction that provides, and other reaction scheme that is fit to is known for those skilled in the art.

D) blocking group.

In some embodiments, the one or more R groups on component aminoacid and/or end amino acid seal with blocking group, most preferably hydrophobic blocking group.Be not limited to specific theory, discovery of the present invention is, sealing, and the sealing that is tried the amino and/or the carboxyl terminal of peptide particularly of the present invention has improved oral delivery widely and has improved serum half-life significantly.

Most blocking groups are suitable for this purpose.This group is including, but not limited to acetyl group, amide and hydrocarbyl group, and terminal protection is particularly preferred for N-for acetyl group and hydrocarbyl group, and protection is preferred to amide group for carboxyl terminal.In some embodiments, described blocking groups can serve as detectable labelling (for example, N-methylamino o-tolyl) in addition.

In some particularly preferred embodiment, described blocking group is including, but not limited to the alkyl chain in the fatty acid, propiono, formoxyl, or the like.Particularly preferred carboxy protective group comprises amide, ester and becomes ether protective group.One preferred embodiment in, acetyl group is used to protect amino terminal, amide groups is used to protect carboxyl terminal.These blocking groups have strengthened the one-tenth spiral trend of described peptide.Some particularly preferred blocking group comprises the alkyl of all lengths, for example, has formula: the group of CH3-(CH2) n-CO-, and wherein n is from about 3 to about 20, and preferably from about 3 to about 16, preferred from 3 to about 13 and most preferred from about 3 to about 10.

Other blocking group comprises; but be not limited to; N-methylamino o-tolyl; Fmoc; tertbutyloxycarbonyl (Boc); 9-fluorenes Acetyl Groups; 1-fluorenes carboxylic group; 9-fluorenes carboxylic group; 9-Fluorenone-1-carboxylic group; benzyloxycarbonyl; xanthyl (xanthine); trityl (Trt); 4-methyl trityl (Mtt); 4-methoxyl group trityl (Mmt); 4-methoxyl group-2; 3; 6-trimethyl-benzenesulfonyl (Mtr); mesitylene-2-sulfonyl (Mts); 4; 4-dimethoxy benzhydryl (Mbh); tosyl (Tos); 2; 2; 5; 7; 8-pentamethyl chromane-6-sulfonyl (Pmc); 4-methylbenzyl (MeBzl); 4-methoxybenzyl (MeOBzl); benzyloxy (BzlO); benzyl (Bzl); benzoyl (Bz); 3-nitro-2-pyridine sulfinyl (Npys); 1-(4; 4-dimethyl-2; 6-dioxy ring caproic subunit) ethyl (Dde); 2; 6-dichloro-benzenes methyl (2,6-DiCl-Bzl); 2-chlorine benzyloxycarbonyl (2-Cl-Z); 2-bromo-benzyloxy-carbonyl (2-Br-Z); benzyloxymethyl (Bom); cyclohexyloxy (cHxO); tert-butoxy methyl (Bum); tert-butoxy (tBuO); the tert-butyl group (tBu); acetyl group (Ac) and trifluoroacetic acid base (TFA).

Protection/blocking groups is known for those skilled in the art, the method that such group is coupled to the suitable residue that comprises peptide of the present invention also be well known to a person skilled in the art (referring to, Greene et al. for example, (1991) Protective Groups in Organic Synthesis, 2nd ed., JohnWiley ﹠amp; Sons, Inc.Somerset, N.J.).One preferred embodiment in, for example, when peptide is on resin, between synthesis stage, use acetic anhydride to realize acetylation.The amide protection can realize by selecting to be used for synthetic suitable resin.During the synthetic in an embodiment peptide described here, use the rink amide resin.Finish synthetic after, at bifunctional acidic aminoacid, the semipermanent blocking group on Asp and Glu and the basic amino acid Lys for example, the semipermanent blocking group on the hydroxyl of Tyr is all removed simultaneously.The peptide that uses acidic treatment to discharge from this resin shows the terminal protected acetyl group that is of N-, and carboxyl is protected to be NH ₂, remove all other blocking group simultaneously.

XI. strengthen the picked-up/oral utilization rate of peptide.

A) the amino acid whose use of D-.

Also be surprising discovery of the present invention be that when with the amino acid whose peptide of D-type (that is, peptide of the present invention) co-administered full L (for example, otherwise having the sequence of peptide of the present invention), the picked-up of D-type peptide has been enhanced.Thereby in some embodiments, the combination of D-type and L-type peptide is used in the present invention's expection in the method for the invention.Described D-type peptide can have different aminoacid sequences with L-type peptide, yet in preferred embodiment, they all have the aminoacid sequence of peptide described here, and in preferred embodiment again, they have identical aminoacid sequence.

Also be discovery of the present invention be that the concatemer of category-A amphiphilic helical peptides of the present invention also is being effective alleviating aspect atherosclerotic one or more symptoms.The monomer that comprises concatemer can directly be coupled at together or connect by joint.In some embodiments, described joint is aminoacid joint (for example, proline), or peptide linker (for example, Gly ₄Ser ₃) (SEQ ID NO:448).In some embodiments, described concatemer is 2 bodies, and 3 bodies more preferably are at more preferably 4 bodies and most preferably 5 bodies, 8 bodies, 10 bodies or 15 bodies.

B) alternative D-and L-aminoacid.

Find, central authorities at peptide replace the hydration that amino acid whose three-dimensional abnormal shape will be allowed particle, and will better allow the lipid of the short struvite oxidation of described peptide particle trapping, fatty acid peroxidase thing and contain the phospholipid of arachidonic oxidation product in the sn-2 position for example.

Thereby, in some embodiments, can synthesize peptide described here to comprise from 4 aminoacid to 10-15 aminoacid, preferred (but dispensable) central authorities (non-end) aminoacid is amino acid whose alternative D and L stereoisomer.End amino acid can be hydrophobic, is because hydrophobic side chain, or because aminoacid have as hydrophobic blocking group described here (for example, the N-end by Boc-, Fmoc-, nicotinoyl-, wait sealing, C-is terminal by (tBu)-sealings such as OtBu).

In table 11, exemplified the example of this peptide.

Table 11. contains some example of the peptide of alternative D-and L-residue at middle section.

Notice that though exemplified concrete aminoacid sequence in table 11, alternative D-and L-aminoacid can be used to any peptide described here.

C) the deutero-peptide of biotin.

In some embodiments, any peptide described here can adhere to one or more biotin (directly or via the indirect covalent coupling of joint).Biotin and enteral sodium dependency multivalence vitamin (multivitamin) transport protein interact, thereby promote the picked-up and the bioavailability of Orally administered peptide.

Biotin can be by any directly coupling of multiple facilitated method well known by persons skilled in the art, or by joint or by amino acid whose side chain coupling.In some embodiments, described biotin is attached to the lysine amino group.

The peptide of biotin that in table 12, exemplified many couplings.

The example of some preferred peptide of table 12.:

XII. pharmaceutical preparation.

In order to carry out method of the present invention, to for example be diagnosed as the individuality that has atherosclerotic one or more symptoms or have atherosclerotic risk use one or more peptides of the present invention or aminoacid to or peptide mimics.Described peptide or aminoacid to or peptide mimics can be with " natural " form, or if desired, use with salt, ester, amide, prodrug, derivant etc., as long as being the pharmacologys, described salt, ester, amide, prodrug or derivant upward be fit to, that is be effective in the methods of the invention.Can use the standard step known to the skilled and for example March (1992) the Advanced Organic Chemistry in synthetic organic chemistry field; Reactions, the step that Mechanisms and Structure, 4th Ed.N.Y.Wiley-Interscience describe prepares salt, ester, amide, prodrug and other derivant of active agent.

For example, use conventional method to prepare acid-addition salts, relate generally to and the acid reaction that is fit to from free alkali.Usually, the alkali form of medicine is dissolved in polar organic solvent, for example in methanol or the ethanol, to wherein adding acid.The salt that produces is precipitated out, or produces from solution by adding lower polar solvent.The acid that is fit to that is used to prepare acid-addition salts comprises organic acid, for example, acetic acid, propanoic acid, hydroxyacetic acid, acetone acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethane sulfonic acid, p-toluenesulfonic acid, salicylic acid, or the like, and mineral acid, for example, hydrochloric acid, hydrobromic acid, sulfacid, nitric acid, phosphoric acid, or the like.Can acid-addition salts be changed into free alkali again by with the alkali treatment that is fit to.The acid-addition salts of particularly preferred activating agent herein is a salt haloid, for example, can use the preparation of hydrochloric acid or hydrobromic acid.On the contrary, can use pharmaceutically acceptable alkali in a similar manner, for example, sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, trimethylamine etc. prepare the goods of the basic salt of described peptide or analogies.Particularly preferred basic salt comprises alkali metal salt, for example, and sodium salt and mantoquita.

The preparation relate generally to hydroxyl of ester and/or the functionalization of carboxylic group, it may reside in the molecular structure of medicine.Described ester usually is free alcohol radical,, derives from the part of the carboxylic acid of formula RCOOH that is, the derivant of acyl substituted, wherein R is an alkyl, preferably lower alkyl.If desired, can ester be changed into free acid again by the hydrogenolysis or the hydrolysing step of routine.

The technology preparation that amide and prodrug also can use the technology of well known to a person skilled in the art or describe in pertinent literature.For example, can use suitable amine reactant to prepare amide from ester, or by preparing them with ammonia or low-grade alkylamine reaction from acid anhydride or acid chloride.Prodrug usually prepares by the covalent attachment of certain part, and this produces a kind of chemical compound, and this chemical compound is that non-activity is gone up in treatment before being modified by individual metabolic system.

This peptide of determining or aminoacid to or analogies for (or suction), rectum or local application parenteral, surperficial, oral, nose, for example by aerosol or wear skin ground, for atherosclerosis and/or its symptom and/or for the preventing and/or treating of this one or more other indications of determining, be useful.Depend on the method for using, ingredient can be used with various unit dosage forms.The unit dosage form that is fit to, including, but not limited to powder, tablet, pill, capsule, lozenge, suppository, paster, nasal spray, injectibles, implantable extended release preparation, lipid complex, or the like.

Described peptide of the present invention and/or aminoacid to and/or peptide mimics usually with pharmaceutically acceptable carrier (excipient) combination to form pharmacological compositions.Pharmaceutically acceptable carrier can contain one or more physiologys and go up acceptable chemical compound, and it works, for example, and stable composition, or the absorption of raising or reduction active agent.The last acceptable chemical compound of physiology can comprise; for example; carbohydrate, for example glucose, sucrose or glucosan, antioxidant; for example ascorbic acid or glutathion; chelating agen, low molecular weight protein (LMWP), protection and picked-up reinforcing agent be lipid for example; reduce the removing of active agent or the compositions of hydrolysis, or excipient or other stabilizing agent and/or buffer.

Other physiology goes up that acceptable chemical compound comprises wetting agent, emulsifying agent, dispersant or for the growth of prophylaxis of microbial or act on useful especially antiseptic.Various antiseptic are known, comprise, for example phenol and ascorbic acid.One skilled in the art will appreciate that pharmaceutically acceptable carrier, comprise that the selection of the last acceptable chemical compound of physiology is depended on, for example, the individually defined thing physical chemistry characteristic of the route of administration of active agent and active agent.

Excipient is preferably aseptic, does not usually contain undesirable material.This compositions can be sterilized by sterilization technology routine, known.

In therapeutic is used, with enough healings or the prevention or the quantity of retardation disease and/or its complication at least in part, use compositions of the present invention to patient, described patient has atherosclerotic one or more symptoms, or has atherosclerotic risk.The enough quantity that achieves this end is defined as " treatment effective dose ".Depend on the severity of disease and the general state of patient health state for the effective quantity of this purposes.The single or multiple of compositions is used and is depended on required dosage and frequency, and patient's tolerance level.In any case described compositions should provide the activating agent of the preparation of the present invention of sufficient amount to treat (improving one or more symptoms) patient effectively.

Peptide or aminoacid to or the concentration of analogies can change widely, mainly based on liquid volume, viscosity, body weight etc., select according to selected specific application form and patient's demand.Yet, usually select concentration so that scope about 0.1 or 1mg/kg/ days to about 50mg/kg/ days and higher sometimes dosage to be provided.Typical dosage range was from about 3mg/kg/ days to about 3.5mg/kg/ days, preferably from about 3.5mg/kg/ days to about 7.2mg/kg/ days, preferred from about 7.2mg/kg/ days to about 11.0mg/kg/ days with most preferred from about 11.0mg/kg/ days to about 15.0mg/kg/ days.Some preferred embodiment in, dosage range was from about 10mg/kg/ days to about 50mg/kg/ days.Should be understood that this dosage can change to optimize the treatment taking method in particular subject or experimenter's group.

Some preferred embodiment in, peptide of the present invention and/or aminoacid to and/or peptide mimics be according to well known to a person skilled in the art standard method oral (for example, passing through tablet) or using as injection.Other preferred embodiment in, described peptide or aminoacid are sent also using the conventional drug delivery system of wearing epidermis, promptly, wear " paster (patches) " of epidermis, wherein active agent generally is comprised in layer structure inside, and layer structure has been served as the drug delivery device that is attached to skin.In this structure, pharmaceutical composition usually is comprised in layer or is arranged in " bank (reservoir) " under the bed course of top.Should be understood that term in context " bank " is meant a certain amount of " active component " that finally can be delivered to skin surface.Thereby for example, " bank " can comprise the active component on the bed course that sticks to described paster, or the active component in well known to a person skilled in the art various different substrates preparations any.Described paster can contain single bank, maybe can contain a plurality of banks.

In one embodiment, described bank comprises the polymeric matrices of pharmaceutically acceptable contact jointing material, and it is used between the medicine delivery period this system being attached on the skin.The example of the contact skin jointing material that is fit to including, but not limited to, polyethylene, polysiloxanes, polyisobutylene, polyacrylate, polyurethanes, or the like.Alternatively, the bank and the contact skin binding agent that contain medicine exist independently and with isolating layer, and binding agent is under bank, in this case, can be aforesaid polymeric matrices, maybe can be the bank of liquid or hydrogel, maybe can adopt other form.Bed course in these are stacked has played the effect of the upper surface of device, preferably plays the effect of the infrastructure components of described " paster ", provides it flexible greatly to described device.The material of selecting for described bed course is impermeable basically for any other material of described active agent and existence preferably.

Be used for other preferred preparation that surface drug sends including, but not limited to ointment and ointment.Ointment is semisolid goods, usually based on vaseline or other petroleum derivative.The ointment that contains selected active agent usually is viscous liquid or semi-solid Emulsion, usually is sensing water content mixed in oil (oil-in-water) or water-in-oil (W-O) (water-in-oil).The ointment cream base generally can be washed, and contains oil phase, emulsifying agent and water.Described oil phase is also referred to as " inside " phase sometimes, and usually for example hexadecane or octadecanol are formed by vaseline and aliphatic alcohol; Although and water is not certain usually, on volume, surpasses oil phase, and usually contain wetting agent.Emulsifying agent in the ointment preparation usually is non-ionic, anionic, cationic or amphoteric surfactant.As will be understood by the skilled person in the art, the cream base of the concrete ointment of use or ointment is used for the optimization medicine and sends.The same with other carrier or excipient, the ointment cream base should be inert, stable, nonirritant and no hypersensitive.

Be different from typical peptide formulations, comprise amino acid whose peptide of the present invention of D-type or aminoacid in addition can use orally, need not protect at the proteolysis of gastric acid, or the like.However, in some embodiments, sending of peptide can strengthen by using the protectiveness excipient.Usually by with polypeptide and compositions is compound makes it to acid or enzyme hydrolysis effect resistance be arranged, or suitably resist carrier, for example in the liposome, realize this point by polypeptide is encapsulated into.For the method for oral delivery protection polypeptide be well known in the art (referring to, for example, United States Patent (USP) 5,391,377 has been described the lipid composition that is used for oral delivery treatment reagent).

A) sustained release formulation.

Can keep the serum half-life of rising by using albumen " encapsulation " system that continues to discharge.This sustained release system is well known to a person skilled in the art.One preferred embodiment in, be used for the biodegradable microsphere delivery system of protease (Tracy (1998) Biotechnol.Prog.14:108 of albumen and peptide; Johnson et al. (1996), Nature Med.2:795; Herbertet al. (1998), Pharmaceut.Res.15,357), a kind ofly have or not combined with the drying agent of other reagent by in polymeric matrix, containing the dried powder that proteic Biodegradable polymeric microsphere is formed, can be used as.

Designing protease microsphere manufacture process especially reaches the albumen sealing effectiveness of height and keeps proteic integrity.Described process is made up of following: (1) has the drug solution of stabilising carriers from the freeze dried albumen particle goods of (bulk) in bulk protein Preparation by the spraying lyophilizing, (ii) prepare the drug-polymer suspension, reduce drug particle size by supersound process or homogenization subsequently, (iii) produce refrigerated drug-polymer microsphere in the liquid nitrogen by being atomised to, (iv) use the ethanol extraction polymer solvent and (v) filter and vacuum drying to produce last dried powder product.The powder that produces contains the albumen of solid form, and it is distributed in the porous polymer particles equably and strictly.Be most commonly used to the polymer of this process, poly-(lactide-co-glycolide) is biocompatible and biodegradable (PLG).

Encapsulation can realize (for example ,-40 ℃) at low temperatures.During encapsulating, albumen is maintained at the solid state that does not have water, thereby the inductive protein conformation transmutability of water is minimized, and prevents to comprise that water is as the degradation reaction of reactant with avoided the organic-aqueous interface that albumen therein may experience degeneration.Preferred process has used most of albumen can not be dissolved in wherein solvent, thereby has produced high encapsulation and render a service (for example, greater than 95%).

In another embodiment, can be used as " concentrate (concentrate) " one or more components of solution are provided, for example, at the preservation container of preparing to be used to dilute (for example, by the volume of measuring in advance) in, or in preparing the solubilized capsule that in the water of certain volume, adds.

B) Zu He preparation.

In some cases, unite that one or more active agents (for example, inhibin, Beta receptor blockers, ACE mortifier, lipid, or the like) are used one or more peptides of the present invention and/or aminoacid is right.Two kinds of reagent (for example, peptide and inhibin) can be side by side or order use.When order when using, use two kinds of reagent like this, thus similar period two kinds of reagent all reach corresponding concentration on the physiology (for example, thereby two kinds of reagent are activated in certain common time).

In some embodiments, side by side use two kinds of reagent.In this case, providing two kinds of reagent in the preparation of single combination is easily.This can realize by well known to a person skilled in the art the whole bag of tricks.For example, in tablet formulation, described tablet can comprise two-layer, and one deck comprises for example inhibin, and another layer comprises for example peptide.In the capsule that regularly discharges, capsule can comprise two pearl devices that regularly discharge, and one is used for peptide, and one contains inhibin.

Above-mentioned preparation and application process are illustrative and nonrestrictive.Should be understood that, use, can easily design preparation and mode of administration that other is fit in this instruction that provides.

XIII. other pharmacological activity reagent.

Other pharmacology go up active agent can with main active agent of the present invention, for example peptide or aminoacid are to together sending.This in one embodiment medicament is including, but not limited to the medicament of the risk that reduces atherosclerotic event and/or its complication.This medicament is including, but not limited to combination, inhibin, aspirin, ace mortifier, the ace receptor inhibitor (ARBs) of Beta receptor blockers, Beta receptor blockers and thiazide diuretic, or the like.

A) inhibin.

Surprising discovery is that " simultaneously " uses one or more peptides of the present invention with one or more inhibin, strengthens the effect of inhibin synergistically.That is to say that inhibin can reach similar effectiveness with lower dosage, thereby avoided the potential adverse side effect (for example, muscle consumption) relevant, and/or impel inhibin significantly to strengthen at any given dosage anti-inflammatory with these medicines.

The main effect of inhibin is to reduce the LDL-cholesterol levels, and they more can reduce the LDL-cholesterol than the medicine of many other kinds.Inhibin is inhibitory enzyme usually, HMG-CoA reductase, the ratio that it produces at health inner control cholesterol.The generally generation by the cholesterol that slows down and remove the ability of the LDL-cholesterol in blood and come cholesterol reducing of these medicines by improving liver.

As if these medicines greatly reduce T-CHOL and LDL-cholesterol, and this greatly reduces heart attack and deaths from heart disease.Because in these researchs their tracking is noted down and their abilities aspect reduction LDL-cholesterol, when patient needed anticholesteremic agent, inhibin had become the most normal medicine of opening in the prescription.Use the research of inhibin reported these medicines in patient percent 20 to 60 lower LDL-cholesterol levels.Inhibin also reduces the triglyceride levels of rising and produces the appropriateness raising of HDL-cholesterol aspect.Recently recognize that inhibin has antiinflammatory property, it does not have direct relation with the lipid reduction degree that realizes.For example, have been found that inhibin reduces the blood plasma level of struvite mark CRP, relatively irrelevant with the variation of the horizontal aspect of blood plasma lipide.Have been found that this anti-inflammatory activity of inhibin, compare, predicting aspect the inductive clinical events minimizing of inhibin no less important or more important with the degree that LDL reduces.

Usually give inhibin at supper or before sleeping by single dose.These medicines are administration at night usually, utilizes this fact, and health is made more cholesterol at night than the daytime.When making up with peptide described here, the peptide of combination/inhibin treatment dosage regimen is usually also carried out at night.

The inhibin that is fit to is well known to a person skilled in the art.This inhibin including, but not limited to torvastatin (

Pfizer), simvastatin ( Merck), pravastatin ( Fluvastatin (

Novartis), lovastatin ( Merck), rosuvastatin (

Astra Zeneca) and Pitavastatin (Sankyo), or the like.

Inhibin/peptide the dosage of combination can carry out conventional optimization to each patient.Usually, inhibin is having maximum effect 4 to 6 weeks in back display result of several weeks.Before with one of inhibin and peptide described here combined therapy, the doctor will obtain conventional test in order to begin inhibin, comprise LDL-cholesterol and HDL-cholesterol levels.In addition, the doctor also will use high-sensitivity detection to measure anti-inflammatory property and the mensuration CRP level of patient's HDL.After the combined therapy in about 4 to 6 weeks, usually doctor's dosage that will repeat these tests and adjust medication realizes that maximum lipid reduces and maximum anti-inflammatory activity.

B) cholesterol absorption mortifier

In some embodiments, unite one or more cholesterol absorption mortifiers to the experimenter use one or more peptides of the present invention and/or aminoacid right.Described peptide can be before, use with the cholesterol absorption mortifier afterwards or side by side.In the later case, described cholesterol absorption mortifier can be used as preparation independently or provides as the preparation with the combination of one or more described peptides.

The cholesterol absorption mortifier is well known to a person skilled in the art.A kind of important cholesterol absorption mortifier is an ezetimibe, is also referred to as 1-(4-fluorophenyl)-3 (R)-[3-(4-fluorophenyl)-3 (S)-hydroxypropyls]-4 (S)-(4-hydroxyphenyl)-2-azetidinone (obtaining from Merck).Ezetimibe reduces blood cholesterol levels by the cholesterol absorption that suppresses small intestinal.

C) Beta receptor blockers.

The Beta receptor blockers that is fit to is including, but not limited to cardioselective (optionally β 1 blocker), for example, and acebutolol (acebutolol, Sectral ^TM), atenolol (atenolol, Tenormin ^TM), betaxolol (betaxolol, Kerlone ^TM), bisoprolol (bisoprolol, Zebeta ^TM), metoprolol (metoprolol, Lopressor ^TM), or the like.The nonselective blocker (equally blocking β 1 and β 2) that is fit to is including, but not limited to carteolol (carteolol, Cartrol ^TM), nadolol (nadolol, Corgard ^TM), penbutolol (penbutolol, Levatol ^TM), pindolol (pindolol, Visken ^TM), carvedilol (carvedilol, Coreg ^TM), Propranolol (propranolol, Inderal ^TM), timolol (timolol, Blockadren ^TM), labetalol (Normodyne ^TM, Trandate ^TM), or the like.

The Beta receptor blockers that is fit to and the combination of thiazide diuretic be including, but not limited to LopressorHCT, ZIAC, Tenoretic, Corzide, Timolide, Inderal LA 40/25, Inderide, Normozide, or the like.

D) ACE mortifier.

The ace mortifier that is fit to including, but not limited to captopril (captopril, for example, the Capoten of Squibb ^TM), benazepril (benazepril, for example, the Lotensin of Novartis ^TM), enalapril (enalapril, for example, the Vasotec of Merck ^TM), fosinopril (fosinopril, for example, the Monopril of Bristol-Myers ^TM), lisinopril (lisinopril, for example, the Prinivil of Merck ^TMOr the Zestril of Astra-Zeneca ^TM), quinapril (quinapril, for example, the Accupril of Parke-Davis ^TM), ramipril (ramipril, for example, Hoechst Marion Roussel, the Altace of KingPharmaceuticals ^TM), imidapril (imidapril), perindopril erbumine (for example, the Aceon of Rhone-Polenc Rorer ^TM), trandolapril (trandolapril, for example, the Mavik of KnollPharmaceutical ^TM), or the like.The ARBS (Ace receptor blocking agent) that is fit to includes but not limited to, and losartan (losartan, for example, the Cozaar of Merck ^TM), irbesartan (irbesartan, for example, the Avapro of Sanofi ^TM), Candesartan (candesartan, for example, the Atacand of Astra Merck ^TM), valsartan (valsartan, for example, the Diovan of Novartis ^TM) or the like.

E) based on the preparation of lipid.

In some embodiments, unite one or more lipids to the experimenter use peptide of the present invention and/or aminoacid right.Described lipid can be configured to active ingredient and/or protect and/or strengthen the transhipment/picked-up of described peptide as excipient, or they can use with being separated.

Be not limited to specific theory, discovery of the present invention is, the using (for example, Orally administered) and can improve the HDL/LDL ratio significantly of some phospholipid.In addition, the phospholipid that it is believed that some moderate-length is to transport by being different from the process that relates to common lipid transfer.Thereby the phospholipid of using some moderate-length with peptide of the present invention has jointly brought many benefits: they have protected phospholipid to avoid digestion or hydrolysis, and they have improved the peptide picked-up, and they have improved the HDL/LDL ratio.

Described lipid can form the liposome of encapsulation polypeptide of the present invention, and/or they can be compound with described peptide simply/mix.Make liposome and encapsulation compositions and methods and be and well known to a person skilled in the art (referring to, for example, Martin and Papahadjopoulos (1982) J.Biol.Chem., 257:286-288; Papahadjopoulos et al. (1991) Proc.Natl.Acad.Sci.USA, 88:11460-11464; Huang et al. (1992) Cancer Res., 52:6774-6781; Lasic et al. (1992) FEBS Lett., 312:255-258., or the like).

The preferred phospholipid that is used for these methods has the fatty acid of scope from about 4 carbon to about 24 carbon in sn-1 and sn-2 position.Some preferred embodiment in, described fatty acid is saturated.Other preferred embodiment in, described fatty acid can be undersaturated.Various preferred fatty acids have been described in table 13.

Table 13. is used for the sn-1 that uses preferred phospholipid of D polypeptide and/or the preferred fatty acid of sn-2 position.

The number common name IUPAC title of carbon

3:0 propiono Trianoic

4:0 bytyry Tetranoic

5:0 valeryl Pentanoic

6:0 caproyl Hexanoic

7:0 heptanoyl group Heptanoic

8:0 caprylyl Octanoic

9:0 pelargonyl group Nonanoic

10:0 capryl Decanoic

11:0 undecanoyl Undecanoic

12:0 12 carbonic acyl radical Dodecanoic

13:0 tridecanoyl Tridecanoic

14:0 myristoyl Tetradecanoic

15:0 pentadecanoyl Pentadecanoic

16:0 palmitoyl Hexadecanoic

17:0 17 carbonic acyl radical Heptadecanoic

18:0 octadecanoyl Octadecanoic

19:0 19 carbonic acyl radical Nonadecanoic

20:0 20 carbonic acyl radical Eicosanoic

21:0 undecanoyl Heniecosanoic

22:0 22 carbonic acyl radical Docosanoic

23:0 two tridecanoyl Trocosanoic

24:0 tetracosa carbon acyl group Tetracosanoic

14:1 is suitable-9-tetradecene acyl group (9-cis)

14:1 is anti--9-tetradecene acyl group (9-trans)

16:1 is suitable-9-hexadecene acyl group (9-cis)

16:1 is anti--9-hexadecene acyl group (9-trans)

Fatty acid in these positions can be identical or different.Particularly preferred phospholipid has phosphocholine in the sn-3 position.

XIV. test kit.

Provide in yet another embodiment of the present invention and be used to improve atherosclerotic one or more symptoms, and/or be used to have the experimenter's (human or animal) of atherosclerosis risk prophylactic treatment, and/or before being used to stimulate-formation and the circulation of β high density lipoprotein like-particles, and/or be used to suppress the test kit of one or more symptoms of osteoporosis.Described test kit preferably comprise contain one or more described peptides of the present invention and/or aminoacid to and/or the container of peptide mimics.Described peptide and/or aminoacid to and/or peptide mimics can provide with unit dose formulations (for example, suppository, tablet, ingot, paster, or the like), and/or optional can with one or more pharmaceutically acceptable excipient composition.

Choose wantonly, described test kit can further comprise and be used for the treatment of heart disease and/or atherosclerotic one or more other reagent.This reagent including, but not limited to, for example aforesaid, Beta receptor blockers, vasodilation, aspirin, inhibin, ace mortifier or ace receptor inhibitor (ARB) etc.

Some preferred embodiment in, described test kit (for example comprises individually inhibin preparation or that be mixed with the formulated in combination form with described peptide in addition, cerivastatin, atorvastatin, simvastatin, pravastatin, fluvastatin, lovastatin, rosuvastatin, Pitavastatin, or the like).Usually, the dosage of inhibin can be lower than the inhibin dosage of generally prescribing under the situation that does not have the concertedness peptide in this preparation.

In addition, described test kit is optional comprises label and/or guiding material, and the guidance (that is scheme) to the use of the practical operation of method or " treatment " of the present invention or " prevention " is provided.Preferred guiding material has been described and has been used one or more polypeptide of the present invention and/or aminoacid to alleviating atherosclerotic one or more symptoms, and/or the outbreak or the increase of prevention one or more this symptoms in the individuality that the atherosclerosis risk is arranged, and/or before stimulating-formation and the circulation of β high density lipoprotein like-particles, and/or one or more symptoms of inhibition osteoporosis, and/or alleviation is one or more symptoms of the pathological changes of feature with the inflammatory reaction.Described guiding material is also passable, optional, instruction preferred dosage/treatment taking method, anti-indication (counter indications) or the like.

Though these guiding materials usually comprise hand-written or materials printed, they are not limited thereto.Can store this explanation and all be that the present invention expects any media that they are communicated to the terminal use.This media is including, but not limited to electronics storage medium (for example, disk, tape, cassette tape, chip), optical medium (for example, CD ROM) or the like.This media can comprise provides the Internet of this guiding material dot address.

Embodiment

Following examples explanation is provided, rather than limits invention required for protection.

Embodiment 1

Assessment to the symptom of little peptide-mediated atherosclerosis and other struvite pathological changes.

Apo A-I analogies described here (referring to, for example, table 1) shows the atherosclerosis characteristic that is similar to apo A-I, because they (have for example removed arterial wall cell oxidation LDL required " seed molecule ", the phospholipid of oxidation is Ox-PAPC, POVPC, PGPC and PEIPC for example, or the like), and be similar to apo A-I, because they have improved atherosclerosis in mouse model.

Apo A-I simulating peptide (for example, D-4F, SEQ ID NO:8) be different from apo A-I be because, they be similar to apo J in hatching altogether also be activated (referring to, for example, USSN10/120,508 and PCT/US03/09988).The general DIYU apo A-I of these peptides does not have sequence homology basically, but they helical structure and they aspect the ability of lipid, have homology.

Less peptide described here (referring to, for example, table 4-7 herein) is similar to natural apoA-I, be because they prevent the LDL oxidation and with the preincubate (pre-incubation) of arterial wall cell rather than hatch altogether prevent in (co-incubation) the inductive monocyte chemotactic activity of LDL (referring to, for example, accompanying drawing 3).

The peptide of describing in accompanying drawing 3 also is activated (accompanying drawing 4) in vivo.Tetrapeptide or D-4F (SEQ ID NO:8) are added in the drinking water of apoE nude mouse (mouse model that the human artery is atherosis) with 5 μ g/mL, or do not add in the drinking water.To the mice blood-letting, separate their lipoprotein by FPLC after 18 hours.Interpolation from the FPLC fraction after the fraction mice of having accepted no peptide drinking water, that contain sophisticated HDL or these fractions (before wherein expection has-β HDL, the i.e. particle under the FPLC post at once after main HDL peak; After the HDL), improved by the inductive monocyte chemotactic activity of the contrast LDL that is added to human artery parietal cell coculture (accompanying drawing 4).In contrast, the FPLC fraction has reduced the inductive monocyte chemotactic activity of LDL significantly after adding to have accepted in the comfortable drinking water HDL of mice of tetrapeptide or D-4F or HDL, shows that described tetrapeptide and D-4F are transformed into anti-inflammatory state (accompanying drawing 4) with these lipoproteins from short inflammatory states.

As shown in Figure 5, take from the LDL of the mice of having accepted described tetrapeptide or D-4F, compare with LDL from the mice of not accepting described peptide, inductive monocyte chemotactic is active significantly to be reduced, and has confirmed the biological activity of Orally administered D-tetrapeptide.

Accompanying drawing 6 shows, be instilled into by stomach tube in the stomach of apoE nude mouse after 20 minutes or 6 hours at SEQ ID NO:258 the synthetic table 4 of D-aminoacid, the HDL that gathers is from the short struvite anti-inflammatory that is converted into, and be similar to HDL from the mice of having accepted D-4F, be different from HDL fully from the mice of having accepted a kind of peptide, the same among this peptide and the D-4F have identical D-aminoacid, thereby but arrange in a certain way with the formation that prevents category-A amphiphilic spiral and make this peptide can not be in conjunction with lipid (mixed and disorderly D-4F).

Accompanying drawing 7 shows, Orally administered by synthetic D-4F of D-aminoacid or SEQ IDNO:258 after mice LDL after 20 minutes or 6 hours, compare with the LDL that takes from the mice of accepting described mixed and disorderly D-4F peptide, induce that monocyte chemotactic is active significantly to be reduced.

Accompanying drawing 8 shows, adds the SEQ ID NO:238 (synthetic by D-aminoacid entirely) 18 hours of table 4 in the food of apoE nude mouse, and the short struvite HDL of apoE nude mouse is changed into anti-inflammatory HDL.

Accompanying drawing 9 shows, at external described tetrapeptide (the SEQ ID NO:258 in the table 4) than effective ten times of SEQID NO:238.

As shown in Figure 3, the SEQ ID NO:238 of 125 μ g/mL only is that moderate is effective, and shown in the accompanying drawing 9, the SEQ ID NO:258 of 12.5 μ g/mL is highly effective in external preincubate.

The experiment that shows in accompanying drawing 10 shows that SEQ ID NO:243, the SEQ IDNO:242 of table 4 and SEQ ID NO:256 also can change into anti-inflammatory HDL with the short struvite HDL of apoE nude mouse.

The activity of particular peptide of the present invention depends on the specific amino acids replacement shown in accompanying drawing 11,12 and 13.SEQ ID NO:254 is identical with SEQ ID NO:258, except arginine in sequence and the amino acid whose position of glutamic acid be opposite (promptly, SEQ ID NO:254 is Boc-Lys (ε Boc)-Glu-Arg-Ser (tBu)-OtBu, and SEQ ID NO:258 is Boc-Lys (ε Boc)-Arg-Glu-Ser (tBu)-OtBu).Go up the result of small variation as this surface, SEQ ID NO:254 has lower in fact effect than SEQ ID NO:258 in these detect.

The experiment of describing in accompanying drawing 11 and 12 shows, the SEQ ID NO:258 of table 4 will urge struvite HDL change into anti-inflammatory HDL and LDL reduced induce monocyte chemotactic active aspect, all more more effective than SEQ ID NO:254 or SEQ ID NO:282.

Serum amyloid A protein (SAA) is the positive acute phase reactant in the mice, is similar to the C-reactive protein (CRP) among the mankind.Data in the accompanying drawing 13 show that this acute phase reactant has been considerably reduced in the blood plasma after having injected SEQ IDNO:258, and the degree that reduces behind injection SEQ IDNO:254 and 282 is slightly little, inapparent.

Accompanying drawing 14 shows, the peptide such as the SEQ ID NO:258 that describe in the table 4 by L-aminoacid during synthetic and orally give apoE nude mouse, change into anti-inflammatory with short struvite HDL, and improved blood plasma paraoxonase activity (accompanying drawing 15) when entirely.

Accompanying drawing 16,17,18 and 19 has shown peptide such as the SEQ ID NO:258 that describes in the table 4, when entirely by D-aminoacid during synthetic and orally give apoE nude mouse, cause HDL to have anti-inflammatory (accompanying drawing 16 and 17), reduced the inductive monocyte chemotactic activity of LDL-(accompanying drawing 17) and improved blood plasma HDL-cholesterol (accompanying drawing 18) and improved HDL paraoxonase activity (accompanying drawing 19).These data also show SEQ ID NO:238, when entirely by L-aminoacid during synthetic and orally give apoE nude mouse, do not change the characteristic (accompanying drawing 16 and 17) of HDL significantly at aspect of inflammation, significantly do not change the inductive monocyte chemotactic activity of LDL (accompanying drawing 17) yet, significantly do not change blood plasma HDL-cholesterol concentration (accompanying drawing 18) yet, significantly do not change HDL paraoxonase activity (accompanying drawing 19) yet.In addition, these data show, when the SEQ of table 4 ID NO:238 entirely by D-aminoacid during synthetic and orally give apoE nude mouse, bring anti-inflammatory (accompanying drawing 16 and 17) to HDL, and reduced the inductive monocyte chemotactic activity of LDL (accompanying drawing 17), but these two kinds of variations do not resemble SEQ ID NO:258 remarkable.In addition, be different from SEQ ID NO:258, the SEQ ID NO:238 of table 4 does not improve HDL-cholesterol concentration (accompanying drawing 18) when being synthesized by D-aminoacid entirely, do not improve HDL paraoxonase activity (accompanying drawing 19).We conclude that the SEQ ID NO:238 of table 4 is invalid when oral giving when being synthesized by L-aminoacid, but be effectively when being synthesized by D-aminoacid, but its effectiveness is lower than SEQ ID NO:258 in fact.

Show in these data that present, SEQ ID NO:238 generally is invalid when being given by the synthetic parallel port of L-aminoacid entirely, when being effectively by D-aminoacid when synthetic entirely, but the effectiveness by the synthetic Orally administered SEQ ID NO:258 of D-aminoacid entirely than same dose is low basically.

Embodiment 2

Peptides synergize statin activity

Accompanying drawing 20 and 21 has shown the remarkable synergism that improves between inhibin (pravastatin) and the D-4F aspect the atherosclerosis in the apoE nude mouse.Known mice is to suppressing to have resistance.The mice of having accepted pravastatin with 20 μ g/mL in drinking water has consumed the suitable pravastatin dosage with human 175mg every day of 70kg, and the mice of having accepted pravastatin with 50 μ g/mL in drinking water has consumed the suitable pravastatin dosage with human 437.5mg every day of 70kg.As shown in accompanying drawing 20 and 21, these very high pravastatin dosage do not have effect aspect the atherosclerotic lesion improving in the apoE nude mouse.As shown in accompanying drawing 20 and 21, add D-4F to the drinking water of apoE nude mouse with the concentration of 2 μ g/mL or 5 μ g/mL separately and do not reduce atherosclerotic lesion.These D-4F dosage are equivalent to the dosage of the 70Kg mankind's 17.5mg every day and 43.75mg every day respectively.Significantly, as shown in accompanying drawing 20 and 21, in the drinking water of apoE nude mouse, together add the pravastatin of same concentrations and D-4F and eliminated atherosclerosis in these mices basically.This shows the synergism of the very high level between inhibin (pravastatin) and the D-4F.

Accompanying drawing 22 shown, the SEQ ID NO.198 of table 4 and SEQ ID NO.203 reduce in the apoE nude mouse aspect the lipid peroxide contents of LDL and HDL, and comparing with D-4F has suitable effect or even more effective.These data are consistent with the peptide of D-4F and description in this application, and described peptide moiety ground induces required " the seed molecule " of struvite atherosclerosis reaction to work by isolated LDL.Data presented in the accompanying drawing 3 to 19 is combined, likelyly is, in this application the peptide of Miao Shuing (for example, the SEQ ID NO:250 of table 4,198 with SEQ ID NO:258) improve aspect the atherosclerosis the same with D-4F effective or more effective.

Embodiment 3

Prediction can make the HDL anti-inflammatory stronger mammiferous atherosclerotic novel little with alleviation The physical property of organic molecule (molecular weight＜900 dalton)

Surprising discovery of the present invention is, many physical propertys have been predicted the ability of little peptide of the present invention, can make that the HDL anti-inflammatory is stronger and alleviates mammiferous atherosclerosis and/or be other pathological changes of feature with the inflammatory reaction.Described physical property be included in the ethyl acetate highly dissoluble (for example, greater than about 4mg/mL) and in the dissolubility of the aqueous buffer solution of pH 7.0.At contact phospholipid, for example 1, during two myristoyls of 2--sn-glyceryl-3-phosphocholine (DMPC), in aqueous environments, effective especially little peptide forms the about 7.5nm of diameter (± 0.1nm) particle, and/or form stacked bilayer, bilayer size general 3.4 to 4.1nm, the about 2nm in gap between the bilayer in stacked, and/or also form the loose structure of about 38nm.Some preferred embodiment in, described little peptide has the molecular weight that is lower than about 900Da.

By comparing the prophesy effect that two sequences illustrate these physical propertys:

SEQ ID NO 254:Boc-Lys (ε Boc)-Glu-Arg-Ser (tBu)-OtBu; With

SEQ?ID?NO?258：Boc-Lys(εBoc)-Arg-Glu-Ser(tBu)-OtBu

In order to be evaluated at the dissolubility in the ethyl acetate, every kind of peptide is weighed, and join in the centrifuge tube, add ethyl acetate (HPLC level; Residue after the evaporation＜0.0001%) to obtain the concentration of 10mg/mL.With test tube seal with wax, vortex, kept 30 minutes in room temperature, carried out vortex in per 10 minutes.Then 10,000rpm moves on to centrifugal 5 minutes of test tube in the test tube of weighing in advance with supernatant.Evaporation of acetic acid ethyl ester in argon is weighed to test tube and to be measured the quantity that is included in the peptide in the described supernatant.Being dissolved in the percentage ratio that peptide in the supernatant accounts for the peptide of initial interpolation shows on Y-axis.Data are meansigma methods ± S.D.The false test tube of handling is represented in contrast, and SEQ ID NO 254 and SEQ ID NO 258 are synthetic by D-aminoacid entirely; SEQ ID NO 250 is synthetic by L-aminoacid entirely.

As shown in the accompanying drawing 23, SEQ ID NO 258 very easily is dissolved in ethyl acetate, and SEQID NO 254 is not (being synthetic by D-aminoacid entirely).In addition, the data in the accompanying drawing 23 show, SEQ ID NO 250[Boc-Phe-Arg-Glu-Leu-OtBu] (entirely by L-aminoacid synthetic) also very easily be dissolved in ethyl acetate.

1mg/mL DMPC suspension in phosphate buffered saline (PBS) (PBS) adds 10% dexycholate, dissolves up to DMPC.Add peptide SEQ ID NO 258 or SEQ ID NO 254 (DMPC: peptide; 1: 10; Wt: wt), reactant mixture is dialysed.After the dialysis, remaining clarifying for SEQ ID NO:258 solution, is muddy for SEQ ID NO:254 solution after dexycholate is removed in dialysis still.

Accompanying drawing 24-26 shows, when in aqueous environments, adding SEQ ID NO 258 to DMPC, form the particle of the about 7.5nm of diameter, form stacked lipid bilayer, bilayer size general 3.4 to 4.1nm, the about 2nm in gap between the bilayer in stacked also forms the loose structure of about 38nm.

Especially, accompanying drawing 24 has shown with negative staining electron micrograph preparation and under 147,420 * enlargement ratio.Arrow has been pointed out SEQ ID NO 258 particles, is measured as 7.5nm (they look like little white particles).

As explanation in accompanying drawing 25, under aqueous environments, the peptide that comprises SEQ ID NO 258 is added to the particle (white arrow) of the about 7.5nm of DMPC formation diameter, with stacked lipid peptide bilayer (pointing to the twill arrow of the white line in disk cylindrical stacked), bilayer size general 3.4 is to 4.1nm, the about 2nm in the gap between the bilayer (black line in disk stacked between the white line).

Accompanying drawing 26 has shown that the peptide of the SEQ ID NO 258 that adds DMPC in aqueous environments to forms the loose structure (white arrow) of stacked lipid-peptide bilayer (twill arrow) and about 38nm.

Accompanying drawing 27 has shown that the DMPC that does not have SEQ ID NO 258 in aqueous environments does not form the particle of the about 7.5nm of diameter, or stacked lipid-peptide bilayer, does not also form the loose structure of about 38nm.

Under the condition of in as accompanying drawing 24, describing, the peptide of SEQ ID NO 254 is (for the amino and the carboxyl terminal of peptide, itself and the different orders that only are arginine and glutamic acid of the peptide of SEQ ID NO 258) do not form the particle of the about 7.5nm of diameter or stacked lipid-peptide bilayer, do not form the loose structure (data not shown) of about 38nm yet.Thereby, the arginine in peptide and the order of glutamic acid have changed it and the interactional ability of DMPC significantly, this point is predicted (promptly by the dissolubility in ethyl acetate, the peptide height of SEQ ID NO 258 dissolves in ethyl acetate, and the formation about 7.5nm particle of diameter and stacked lipid-peptide bilayer, and the loose structure of about 38nm, and the peptide of SEQ ID NO 254 is dissolved in ethyl acetate rarely, does not form these structures under the condition under describing as accompanying drawing 24).Except the scheme of in accompanying drawing 24, describing, if the DMPC suspension among the PBS is added to the peptide (DMPC: peptide of SEQ ID NO 258; 1: 10; Wt: wt) or the peptide (DMPC: peptide of SEQ ID NO 254; 1: 10; Wt: wt), between transition temperature that only is higher than DMPC (transition temperature) (only being higher than 50 ℃) and room temperature, per hour several cycles mixture is circulated, be placed on following 48 hours of room temperature (data not shown) then, also obtained similar result.

The physical property of the peptide of SEQ ID NO 258 (rather than peptide of SEQ ID NO 254) shows, this peptide has amphipathic characteristic (promptly, it highly dissolves in ethyl acetate, also dissolves in the aqueous buffer solution [data not shown] of pH 7.0, and what it was as implied above interacts with DMPC).Surprising discovery of the present invention is, highly dissolve in ethyl acetate, and also dissolve in the peptide of the aqueous buffer solution of pH 7.0, form lipid-peptide complexes with the DMPC interaction, it is similar to the nascent HDL particle (Forte, et al. (1993) J.Lipid Res.34:317-324) that the interaction by apoA-I and cell forms significantly.

Table 13 has compared in the interaction of the human apoA-I of external no lipid and CHO-C19 cell and as the interaction of the SEQ ID NO that points out of above accompanying drawing 4-7 258 and DMPC.

The peptide of indicated SEQ ID NO 258 and the interaction of DMPC among table 13. as the above accompanying drawing 24-27, and as the comparison between the interaction of the human apoA-I of the no lipid described among Forte et al. (1993) the J.Lipid Res.34:317-324 and CHO-C-19 cell.

Thereby, highly dissolve in the described here little peptide that ethyl acetate also also dissolves in the aqueous buffer solution of pH 7.0, interact with lipid (DMPC) with apoA-I similarly, apoA-I has 28,000 daltonian molecular weight.

The molecular model that shows among the accompanying drawing 28-32 has shown space characteristics 258 that compare with SEQ ID NO, SEQ ID NO 254.

The molecular model that shows in accompanying drawing 28-32 shows that the peptide of SEQ ID NO 254 and the peptide of SEQID NO 258 all contain polarity and nonpolar part in each molecule, but has spatial diversity on the arranging of the polarity of two kinds of molecules and non-polar component.As result in the difference aspect the spatial arrangement of molecule, aspect the dissolubility (accompanying drawing 23) of two kinds of molecules in ethyl acetate and they and DMPC interaction aspect (accompanying drawing 24-27) there are differences.

Data among the accompanying drawing 33-35 show, the physical property of the peptide of contrast SEQ ID NO 258 and the peptide of SEQ IDNO 254 has predicted that these molecules make that the HDL anti-inflammatory is stronger and alleviate atherosclerotic ability when giving mammal when oral.

Female apoE nude mouse is not having the diet (chow) of interpolation age or accept to contain the foods of following peptide in 8 weeks: 200 μ g/gm SEQ ID NO 254 (+254) or 200 μ g/gm SEQ ID NO258 (+258), they all are synthetic by D-aminoacid entirely.Give the mice blood-letting after 15 weeks, by FPLC fractional distillation blood plasma, their HDL (mHDL) of test in human arterial wall cell coculture.The human LDL of an interpolation standard (the LDL-cholesterol of 100 μ g/mL) (LDL) in human arterial wall coculture, or do not add (do not have add), or the mice HDL that the normal human subject HDL of itself and 50 μ g/mL together adds (hHDL) or itself and 50 μ g/mL together adds (mHDL), measure the monocyte chemotactic activity that produces, on Y-axis, mark and draw.Accompanying drawing 33 shows, after mice is fed SEQ ID NO258, rather than after the SEQ ID NO 254 that feeds, has been endowed anti-inflammatory from the HDL of apoE nude mouse.

As shown in the accompanying drawing 34, the peptide of the peptide of SEQ ID NO 258 rather than SEQ ID NO 254 reduces the atherosclerosis in the aortic root (aortic sinus) of aforesaid apoE nude mouse significantly.Accompanying drawing 35 explanations, SEQ ID NO 258 rather than SEQ ID NO 254 also reduce the atherosclerosis in the aortal en face goods significantly.Accompanying drawing 23 explanation improves atherosclerotic ability by the dissolubility (referring to above accompanying drawing 23) of the synthetic SEQ ID of L-aminoacid NO 250 in the ethyl acetate this molecule that calculated to a nicety entirely in the apoE nude mouse.

Thereby the physical property of these the little peptides described peptide that calculates to a nicety is alleviated atherosclerotic ability in the apoE nude mouse.

Thereby we have instructed little peptide, usually have less than about 900 daltonian molecular weight, highly dissolve in ethyl acetate (greater than about 4mg/mL), and dissolve in the aqueous buffer solution of pH 7.0, when in aqueous environments with phospholipid for example 1, the two myristoyls of 2--when sn-glyceryl-3-phosphocholine (DMPC) contacts, form the particle of the about 7.5nm of diameter, and/or form stacked bilayer, bilayer size general 3.4 to 4.1nm, the about 2nm in gap between the bilayer in stacked, and/or they also form the loose structure of about 38nm, when to administration, making the HDL anti-inflammatory stronger is one or more symptoms of other pathological changes of feature with the alleviation atherosclerosis and with the inflammatory reaction.

Will be understood that, embodiment described here and embodiment only are used for illustrative purpose, according to these various modifications of carrying out or change will be for those skilled in the art provides inspiration, and be included in the scope of the application's spirit and scope and subsidiary claim., quote this paper with their integral body and do reference for various purposes in this all publications, patent and patent application of quoting.

Sequence table

＜110〉The Regents of the University of California (THE REGENTS 0F THE UNIVERSITY 0F CALIF0RNIA)

＜120〉Orally administered small peptides synergize statin activity

<130>407T-911320PC

<140>PCT/US2004/026288

<141>2004-08-10

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<151>2003-08-11

<150>US10/649,378

<151>2003-08-26

<160>465

<170>PatentIn?version?3.3

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<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis, aminoacid can be protection or unprotected D or L type

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is aspartic acid or glutamic acid.

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<222>(2)..(3)

＜223〉Xaa is tryptophan, phenylalanine, alanine, leucine, tyrosine, isoleucine, valine or Alpha-Naphthyl alanine

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<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is lysine or arginine.

<220>

<221>misc_feature

<222>(5)..(5)

＜223〉Xaa is serine, threonine, alanine, glycine or histidine

<220>

<221>misc_feature

<222>(6)..(7)

＜223〉Xaa is tryptophan, phenylalanine, alanine, leucine, tyrosine, isoleucine, valine or Alpha-Naphthyl alanine

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<221>misc_feature

<222>(8)..(8)

＜223〉Xaa is aspartic acid or glutamic acid

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<222>(9)..(9)

＜223〉Xaa is lysine or arginine

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<221>misc_feature

<222>(10)..(11)

＜223〉Xaa is tryptophan, phenylalanine, alanine, leucine, tyrosine, isoleucine, valine or Alpha-Naphthyl alanine

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<222>(12)..(12)

＜223〉Xaa is aspartic acid or glutamic acid.

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<222>(13)..(13)

＜223〉Xaa is lysine or arginine

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<221>misc_feature

<222>(14)..(14)

＜223〉Xaa is tryptophan, phenylalanine, alanine, leucine, tyrosine, isoleucine, valine or Alpha-Naphthyl alanine

<220>

<221>misc_feature

<222>(15)..(15)

＜223〉Xaa is lysine or arginine

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<221>misc_feature

<222>(16)..(16)

＜223〉Xaa is aspartic acid or glutamic acid.

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<222>(17)..(18)

＜223〉Xaa is tryptophan, phenylalanine, alanine, leucine, tyrosine, isoleucine, valine or Alpha-Naphthyl alanine

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Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa

1 5 10 15

Xaa?Xaa

<210>2

<211>20

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＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

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<222>(1)..(1)

＜223〉Xaa is Pro, Ala, Gly, Asn, Gln, or D-Pro

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<222>(2)..(2)

＜223〉Xaa is an aliphatic amino acid

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<222>(3)..(3)

＜223〉Xaa is Leu

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<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is an acidic amino acid

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<221>misc_feature

<222>(5)..(6)

＜223〉Xaa is Leu, or Phe

<220>

<221>misc_feature

<222>(7)..(7)

＜223〉Xaa is a basic amino acid

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<221>misc_feature

<222>(8)..(8)

＜223〉Xaa is an acidic amino acid

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<221>misc_feature

<222>(9)..(10)

＜223〉Xaa is Leu, or Trp

<220>

<221>misc_feature

<222>(11)..(11)

＜223〉Xaa is acidic amino acid or Asn

<220>

<221>misc_feature

<222>(12)..(12)

＜223〉Xaa is an acidic amino acid

<220>

<221>misc_feature

<222>(13)..(13)

＜223〉Xaa is Leu, Trp, or Phe

<220>

<221>misc_feature

<222>(14)..(14)

＜223〉Xaa is basic amino acid or Leu

<220>

<221>misc_feature

<222>(15)..(15)

＜223〉Xaa is Gln, or Asn

<220>

<221>misc_feature

<222>(16)..(16)

＜223〉Xaa is a basic amino acid

<220>

<221>misc_feature

<222>(17)..(17)

＜223〉Xaa is Leu

<220>

<221>misc_feature

<222>(18)..(18)

＜223〉Xaa is a basic amino acid

<220>

<221>misc_feature

<222>(19)..(20)

＜223〉Xaa can be the aminoacid of any natural generation

<400>2

Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa

1 5 10 15

Xaa?Xaa?Xaa?Xaa

20

<210>3

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>3

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>4

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>4

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>5

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>5

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>6

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>6

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>7

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>7

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>8

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>8

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>9

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>9

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe

<210>10

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>10

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Phe?Phe?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe

<210>11

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>11

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Phe?Phe?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe

<210>12

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>12

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Phe?Phe

<210>13

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>13

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>14

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>14

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Phe?Phe

<210>15

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>15

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe

<210>16

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>16

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe

<210>17

<211>18

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>17

Glu?Trp?Leu?Lys?Leu?Phe?Tyr?Glu?Lys?Val?Leu?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>18

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>18

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>19

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>19

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Phe?Phe

<210>20

<211>18

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>20

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>21

<211>18

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>21

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Phe?Phe

<210>22

<211>18

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>22

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe

<210>23

<211>18

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>23

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe

<210>24

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>24

Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu?Ala?Phe

1 5 10

<210>25

<211>14

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>25

Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu?Ala?Phe

1 5 10

<210>26

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>26

Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu?Ala?Phe

1 5 10

<210>27

<211>14

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>27

Ala?Phe?Tyr?Asp?Lys?Phe?Phe?Glu?Lys?Phe?Lys?Glu?Phe?Phe

1 5 10

<210>28

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>28

Ala?Phe?Tyr?Asp?Lys?Phe?Phe?Glu?Lys?Phe?Lys?Glu?Phe?Phe

1 5 10

<210>29

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>29

Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu?Ala?Phe

1 5 10

<210>30

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>30

Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu?Phe?Phe

1 5 10

<210>31

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>31

Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu?Ala?Phe

1 5 10

<210>32

<211>14

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>32

Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Leu?Lys?Glu?Phe?Phe

1 5 10

<210>33

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>33

Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu?Phe?Phe

1 5 10

<210>34

<211>14

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>34

Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu?Phe

1 5 10

<210>35

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>35

Leu?Phe?Tyr?Glu?Lys?Val?Leu?Glu?Lys?Phe?Lys?Glu?Ala?Phe

1 5 10

<210>36

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>36

Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu?Ala?Phe

1 5 10

<210>37

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>37

Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu?Phe?Phe

1 5 10

<210>38

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>38

Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu?Ala?Phe

1 5 10

<210>39

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>39

Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Leu?Lys?Glu?Phe?Phe

1 5 10

<210>40

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>40

Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu?Phe?Phe

1 5 10

<210>41

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>41

Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu?Phe?Phe

1 5 10

<210>42

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>42

Asp?Trp?Leu?Lys?Ala?Leu?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Leu

<210>43

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>43

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Phe?Phe

<210>44

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>44

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Glu?Lys?Phe?Phe?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe

<210>45

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>45

Glu?Trp?Leu?Lys?Ala?Leu?Tyr?Glu?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Leu

<210>46

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>46

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>47

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>47

Glu?Trp?Phe?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Phe?Phe

<210>48

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>48

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe

<210>49

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>49

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Glu?Lys?Phe?Phe?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe

<210>50

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>50

Glu?Trp?Phe?Lys?Ala?Phe?Tyr?Glu?Lys?Phe?Phe?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe

<210>51

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>51

Asp?Phe?Leu?Lys?Ala?Trp?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Trp

<210>52

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>52

Glu?Phe?Leu?Lys?Ala?Trp?Tyr?Glu?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Trp

<210>53

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>53

Asp?Phe?Trp?Lys?Ala?Trp?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Trp?Trp

<210>54

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>54

Glu?Phe?Trp?Lys?Ala?Trp?Tyr?Glu?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Trp?Trp

<210>55

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>55

Asp?Lys?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Trp?Ala?Lys?Glu

1 5 10 15

Ala?Phe

<210>56

<211>18

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>56

Asp?Lys?Trp?Lys?Ala?Val?Tyr?Asp?Lys?Phe?Ala?Glu?Ala?Phe?Lys?Glu

1 5 10 15

Phe?Leu

<210>57

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>57

Glu?Lys?Leu?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Phe?Glu?Trp?Ala?Lys?Glu

1 5 10 15

Ala?Phe

<210>58

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>58

Glu?Lys?Trp?Lys?Ala?Val?Tyr?Glu?Lys?Phe?Ala?Glu?Ala?Phe?Lys?Glu

1 5 10 15

Phe?Leu

<210>59

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>59

Asp?Trp?Leu?Lys?Ala?Phe?Val?Asp?Lys?Phe?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Tyr

<210>60

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>60

Glu?Lys?Trp?Lys?Ala?Val?Tyr?Glu?Lys?Phe?Ala?Glu?Ala?Phe?Lys?Glu

1 5 10 15

Phe?Leu

<210>61

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>61

Asp?Trp?Leu?Lys?Ala?Phe?Val?Tyr?Asp?Lys?Val?Phe?Lys?Leu?Lys?Glu

1 5 10 15

Phe?Phe

<210>62

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>62

Glu?Trp?Leu?Lys?Ala?Phe?Val?Tyr?Glu?Lys?Val?Phe?Lys?Leu?Lys?Glu

1 5 10 15

Phe?Phe

<210>63

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>63

Asp?Trp?Leu?Arg?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>64

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>64

Glu?Trp?Leu?Arg?Ala?Phe?Tyr?Glu?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>65

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>65

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Arg?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>66

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>66

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Glu?Arg?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>67

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>67

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Arg?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>68

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>68

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Ala?Glu?Arg?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>69

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>69

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Arg?Glu

1 5 10 15

Ala?Phe

<210>70

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>70

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Ala?Glu?Lys?Leu?Arg?Glu

1 5 10 15

Ala?Phe

<210>71

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>71

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Arg?Val?Ala?Glu?Arg?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>72

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>72

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Glu?Arg?Val?Ala?Glu?Arg?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>73

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>73

Asp?Trp?Leu?Arg?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Arg?Glu

1 5 10 15

Ala?Phe

<210>74

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>74

Glu?Trp?Leu?Arg?Ala?Phe?Tyr?Glu?Lys?Val?Ala?Glu?Lys?Leu?Arg?Glu

1 5 10 15

Ala?Phe

<210>75

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>75

Asp?Trp?Leu?Arg?Ala?Phe?Tyr?Asp?Arg?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>76

<211>18

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>76

Glu?Trp?Leu?Arg?Ala?Phe?Tyr?Glu?Arg?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>77

<211>18

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>77

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Arg?Leu?Arg?Glu

1 5 10 15

Ala?Phe

<210>78

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>78

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Ala?Glu?Arg?Leu?Arg?Glu

1 5 10 15

Ala?Phe

<210>79

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>79

Asp?Trp?Leu?Arg?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Arg?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>80

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>80

Glu?Trp?Leu?Arg?Ala?Phe?Tyr?Glu?Lys?Val?Ala?Glu?Arg?Leu?Lys?Glu

1 5 10 15

Ala?Phe

<210>81

<211>37

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>81

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Phe?Pro?Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys

20 25 30

Leu?Lys?Glu?Ala?Phe

35

<210>82

<211>37

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>82

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Phe?Phe?Pro?Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys

20 25 30

Leu?Lys?Glu?Phe?Phe

35

<210>83

<211>37

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>83

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Leu?Lys?Glu

1 5 10 15

Ala?Phe?Pro?Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys

20 25 30

Leu?Lys?Glu?Ala?Phe

35

<210>84

<211>37

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>84

Asp?Lys?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Trp?Ala?Lys?Glu

1 5 10 15

Ala?Phe?Pro?Asp?Lys?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Trp

20 25 30

Leu?Lys?Glu?Ala?Phe

35

<210>85

<211>37

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>85

Asp?Lys?Trp?Lys?Ala?Val?Tyr?Asp?Lys?Phe?Ala?Glu?Ala?Phe?Lys?Glu

1 5 10 15

Phe?Leu?Pro?Asp?Lys?Trp?Lys?Ala?Val?Tyr?Asp?Lys?Phe?Ala?Glu?Ala

20 25 30

Phe?Lys?Glu?Phe?Leu

35

<210>86

<211>37

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>86

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe?Pro?Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys

20 25 30

Phe?Lys?Glu?Ala?Phe

35

<210>87

<211>37

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>87

Asp?Trp?Leu?Lys?Ala?Phe?Val?Tyr?Asp?Lys?Val?Phe?Lys?Leu?Lys?Glu

1 5 10 15

Phe?Phe?Pro?Asp?Trp?Leu?Lys?Ala?Phe?Val?Tyr?Asp?Lys?Val?Phe?Lys

20 25 30

Leu?Lys?Glu?Phe?Phe

35

<210>88

<211>37

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>88

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Phe?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe?Pro?Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Phe?Ala?Glu?Lys

20 25 30

Phe?Lys?Glu?Phe?Phe

35

<210>89

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>89

Glu?Trp?Phe?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>90

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>90

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe

1 5 10

<210>91

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>91

Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10

<210>92

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>92

Phe?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10

<210>93

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>93

Asn?Met?Ala?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys

1 5 10 15

Glu

<210>94

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>94

Asn?Met?Ala?Phe?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Ala?Glu?Lys?Phe?Lys

1 5 10 15

Glu

<210>95

<211>21

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>95

Asn?Met?Ala?Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys

1 5 10 15

Phe?Lys?Glu?Ala?Phe

20

<210>96

<211>21

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>96

Asn?Met?Ala?Glu?Trp?Phe?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Ala?Glu?Lys

1 5 10 15

Phe?Lys?Glu?Ala?Phe

20

<210>97

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>97

Asn?Met?Ala?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu?Ala

1 5 10 15

Phe

<210>98

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>98

Asn?Met?Ala?Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys

1 5 10 15

Phe

<210>99

<211>39

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>99

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe?Asn?Met?Ala?Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe

20 25 30

Glu?Lys?Phe?Lys?Glu?Phe?Phe

35

<210>100

<211>39

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>100

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Phe?Phe?Asn?Met?Ala?Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Phe

20 25 30

Glu?Lys?Phe?Lys?Glu?Phe?Phe

35

<210>101

<211>31

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>101

Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu?Phe?Phe?Asn?Met

1 5 10 15

Ala?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu?Phe?Phe

20 25 30

<210>102

<211>31

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>102

Ala?Phe?Tyr?Glu?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu?Phe?Phe?Asn?Met

1 5 10 15

Ala?Ala?Phe?Tyr?Glu?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu?Phe?Phe

20 25 30

<210>103

<211>31

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>103

Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Asn?Met

1 5 10 15

Ala?Asp?Trp?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe

20 25 30

<210>104

<211>31

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>104

Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Phe?Glu?Lys?Phe?Asn?Met

1 5 10 15

Ala?Glu?Trp?Leu?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Phe?Glu?Lys?Phe

20 25 30

<210>105

<211>31

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>105

Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu?Asn?Met

1 5 10 15

Ala?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu

20 25 30

<210>106

<211>31

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>106

Leu?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu?Asn?Met

1 5 10 15

Ala?Leu?Lys?Ala?Phe?Tyr?Glu?Lys?Val?Phe?Glu?Lys?Phe?Lys?Glu

20 25 30

<210>107

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>107

Asp?Lys?Trp?Lys?Ala?Val?Tyr?Asp?Lys?Phe?Ala?Glu?Ala?Phe?Lys?Glu

1 5 10 15

Phe?Leu

<210>108

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>108

Asp?Lys?Leu?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Phe?Glu?Trp?Ala?Lys?Glu

1 5 10 15

Ala?Phe

<210>109

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>109

Lys?Arg?Ser

1

<210>110

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>110

Lys?Arg?Thr

1

<210>111

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>111

Trp?Arg?Ile

1

<210>112

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>112

Trp?Arg?Leu

1

<210>113

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>113

Phe?Arg?Ile

1

<210>114

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>114

Phe?Arg?Leu

1

<210>115

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>115

Lys?Glu?Ser

1

<210>116

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>116

Lys?Glu?Thr

1

<210>117

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>117

Lys?Asp?Ser

1

<210>118

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>118

Lys?Asp?Thr

1

<210>119

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>119

Lys?Arg?Ser

1

<210>120

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>120

Lys?Arg?Thr

1

<210>121

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>121

Leu?Glu?Ser

1

<210>122

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>122

Leu?Glu?Thr

1

<210>123

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>123

Trp?Arg?Ser

1

<210>124

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>124

Trp?Asp?Ser

1

<210>125

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>125

Trp?Glu?Ser

1

<210>126

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>126

Trp?Arg?Ser

1

<210>127

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>127

Lys?Glu?Leu

1

<210>128

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>128

Leu?Arg?Ser

1

<210>129

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>129

Leu?Asp?Ser

1

<210>130

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>130

Leu?Glu?Ser

1

<210>131

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>131

Leu?Arg?Ser

1

<210>132

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>132

Leu?Arg?Thr

1

<210>133

<211>3

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>133

Glu?Asp?Tyr

1

<210>134

<211>3

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>134

Lys?Arg?Ser

1

<210>135

<211>3

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>135

Trp?Arg?Ile

1

<210>136

<211>3

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>136

Trp?Arg?Leu

1

<210>137

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>137

Phe?Arg?Ile

1

<210>138

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>138

Phe?Arg?Leu

1

<210>139

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>139

Trp?Arg?Phe

1

<210>140

<211>3

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>140

Trp?Arg?Tyr

1

<210>141

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>141

Trp?Arg?Phe

1

<210>142

<211>3

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>142

Trp?Arg?Tyr

1

<210>143

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine

<400>143

Xaa?Arg?Ser

1

<210>144

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>144

Lys?Arg?Ser

1

<210>145

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>145

Lys?Arg?Thr

1

<210>146

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>146

Leu?Asp?Thr

1

<210>147

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>147

Leu?Glu?Thr

1

<210>148

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>148

Leu?Arg?Thr

1

<210>149

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>149

Xaa?Arg?Ser

1

<210>150

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>150

Xaa?Asp?Ser

1

<210>151

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

<223>Xaa?is?ornithiine

<400>151

Xaa?Glu?Ser

1

<210>152

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>152

Lys?Arg?Ser

1

<210>153

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>153

Lys?Arg?Thr

1

<210>154

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>154

Lys?Glu?Ser

1

<210>155

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>155

Lys?Glu?Thr

1

<210>156

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>156

Lys?Asp?Ser

1

<210>157

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>157

Lys?Asp?Thr

1

<210>158

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>158

Lys?Glu?Leu

1

<210>159

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>159

Lys?Arg?Leu

1

<210>160

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>160

Lys?Arg?Thr

1

<210>161

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>161

Lys?Glu?Ser

1

<210>162

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>162

Lys?Glu?Thr

1

<210>163

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>163

Lys?Asp?Ser

1

<210>164

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>164

Lys?Asp?Thr

1

<210>165

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>165

Lys?Arg?Ser

1

<210>166

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>166

Lys?Glu?Leu

1

<210>167

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>167

Lys?Asp?Ser

1

<210>168

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>168

Lys?Asp?Thr

1

<210>169

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>169

Lys?Arg?Thr

1

<210>170

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>170

Lys?Glu?Leu

1

<210>171

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>171

Xaa?Glu?Ser

1

<210>172

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

<223>Xaa?is?ornithine.

<400>172

Xaa?Asp?Ser

1

<210>173

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>173

Xaa?Asp?Thr

1

<210>174

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>174

Xaa?Arg?Thr

1

<210>175

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>175

Xaa?Glu?Thr

1

<210>176

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>176

Trp?Asp?Ile

1

<210>177

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>177

Trp?Arg?Ile

1

<210>178

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>178

Trp?Glu?Ile

1

<210>179

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>179

Trp?Asp?Leu

1

<210>180

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>180

Trp?Glu?Leu

1

<210>181

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>181

Phe?Asp?Ile

1

<210>182

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>182

Phe?Asp?Leu

1

<210>183

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>183

Phe?Glu?Leu

1

<210>184

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>184

Trp?Arg?Phe

1

<210>185

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>185

Trp?Glu?Phe

1

<210>186

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>186

Trp?Asp?Phe

1

<210>187

<211>3

<212>PRT

<213>Artificial?Sequen?Sequencece

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>187

Trp?Asp?Tyr

1

<210>188

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>188

Trp?Arg?Tyr

1

<210>189

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>189

Trp?Glu?Tyr

1

<210>190

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>190

Trp?Arg?Thr

1

<210>191

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>191

Trp?Asp?Thr

1

<210>192

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>192

Trp?Glu?Thr

1

<210>193

<211>3

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(3)..(3)

＜223〉Xaa is a nor-leucine.

<400>193

Phe?Arg?Xaa

1

<210>194

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(3)..(3)

＜223〉Xaa is a nor-leucine.

<400>194

Phe?Glu?Xaa

1

<210>195

<211>3

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(3)..(3)

＜223〉Xaa is a nor-leucine.

<400>195

Phe?Asp?Xaa

1

<210>196

<211>3

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>196

Glu?His?Tyr

1

<210>197

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>197

Leu?His?Ser

1

<210>198

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>198

Leu?His?Thr

1

<210>199

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>199

Lys?His?Ser

1

<210>200

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>200

Lys?His?Thr

1

<210>201

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>201

Lys?His?Leu

1

<210>202

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>202

Lys?His?Ser

1

<210>203

<211>3

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>203

Lys?His?Thr

1

<210>204

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>204

Lys?His?Leu

1

<210>205

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

<223>Xaa?is?orniithine.

<400>205

Xaa?His?Ser

1

<210>206

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

<223>Xaa?is?orniithine.

<400>206

Xaa?His?Thr

1

<210>207

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>207

Phe?His?Ile

1

<210>208

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>208

Phe?His?Leu

1

<210>209

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(3)..(3)

＜223〉Xaa is a nor-leucine.

<400>209

Phe?His?Xaa

1

<210>210

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>210

Phe?Lys?Leu

1

<210>211

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>211

Trp?His?Ile

1

<210>212

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>212

Trp?His?Leu

1

<210>213

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>213

Trp?His?Phe

1

<210>214

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>214

Trp?His?Tyr

1

<210>215

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>215

Phe?Lys?Leu

1

<210>216

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>216

Lys?His?Ser

1

<210>217

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>217

Lys?His?Thr

1

<210>218

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>218

Lys?His?Leu

1

<210>219

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>219

Leu?His?Ser

1

<210>220

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>220

Leu?His?Thr

1

<210>221

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>221

Lys?His?Ser

1

<210>222

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>222

Lys?His?Thr

1

<210>223

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>223

Lys?His?Leu

1

<210>224

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>224

Lys?His?Ser

1

<210>225

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>225

Lys?His?Thr

1

<210>226

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is a nor-leucine.

<400>226

Xaa?His?Ser

1

<210>227

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>227

Phe?His?Ile

1

<210>228

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>228

Phe?His?Leu

1

<210>229

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(3)..(3)

＜223〉Xaa is a nor-leucine.

<400>229

Phe?His?Xaa

1

<210>230

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>230

Trp?His?Ser

1

<210>231

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>231

Trp?His?Ile

1

<210>232

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>232

Trp?His?Leu

1

<210>233

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>233

Trp?His?Phe

1

<210>234

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>234

Trp?His?Tyr

1

<210>235

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>235

Trp?His?Thr

1

<210>236

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>236

Lys?His?Ser

1

<210>237

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>237

Lys?His?Thr

1

<210>238

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>238

Lys?Arg?Asp?Ser

1

<210>239

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>239

Lys?Arg?Asp?Thr

1

<210>240

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>240

Trp?Arg?Asp?Ile

1

<210>241

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>241

Trp?Arg?Asp?Leu

1

<210>242

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>242

Phe?Arg?Asp?Leu

1

<210>243

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>243

Phe?Arg?Asp?Ile

1

<210>244

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>244

Phe?Arg?Asp?Xaa

1

<210>245

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>245

Phe?Arg?Glu?Xaa

1

<210>246

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>246

Phe?Arg?Glu?Ile

1

<210>247

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>247

Phe?Asp?Arg?Ile

1

<210>248

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>248

Phe?Glu?Arg?Ile

1

<210>249

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>249

Phe?Asp?Arg?Leu

1

<210>250

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>250

Phe?Arg?Glu?Leu

1

<210>251

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>251

Phe?Glu?Arg?Leu

1

<210>252

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>252

Phe?Asp?Arg?Xaa

1

<210>253

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>253

Phe?Glu?Arg?Xaa

1

<210>254

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>254

Lys?Glu?Arg?Ser

1

<210>255

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>255

Lys?Glu?Arg?Thr

1

<210>256

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>256

Lys?Asp?Arg?Ser

1

<210>257

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>257

Lys?Asp?Arg?Thr

1

<210>258

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>258

Lys?Arg?Glu?Ser

1

<210>259

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>259

Lys?Arg?Glu?Thr

1

<210>260

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>260

Leu?Glu?Arg?Ser

1

<210>261

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>261

Leu?Glu?Arg?Thr

1

<210>262

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>262

Trp?Arg?Asp?Ser

1

<210>263

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>263

Trp?Asp?Arg?Ser

1

<210>264

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>264

Trp?Glu?Arg?Ser

1

<210>265

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>265

Trp?Arg?Glu?Ser

1

<210>266

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>266

Lys?Glu?Arg?Leu

1

<210>267

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>267

Leu?Arg?Asp?Ser

1

<210>268

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>268

Leu?Asp?Arg?Ser

1

<210>269

<211>4

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>269

Leu?Glu?Arg?Ser

1

<210>270

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>270

Leu?Arg?Glu?Ser

1

<210>271

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>271

Leu?Arg?Asp?Thr

1

<210>272

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>272

Glu?Asp?Arg?Tyr

1

<210>273

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>273

Lys?Arg?Asp?Ser

1

<210>274

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>274

Trp?Arg?Asp?Ile

1

<210>275

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>275

Trp?Arg?Asp?Leu

1

<210>276

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>276

Phe?Arg?Asp?Ile

1

<210>277

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>277

Phe?Arg?Asp?Leu

1

<210>278

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>278

Trp?Arg?Asp?Phe

1

<210>279

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>279

Trp?Arg?Asp?Tyr

1

<210>280

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>280

Trp?Arg?Asp?Phe

1

<210>281

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>281

Trp?Arg?Asp?Tyr

1

<210>282

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(2)

＜223〉Xaa is an ornithine.

<400>282

Xaa?Arg?Glu?Ser

1

<210>283

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>283

Lys?Arg?Asp?Ser

1

<210>284

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>284

Lys?Arg?Asp?Thr

1

<210>285

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>285

Leu?Asp?Arg?Thr

1

<210>286

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>286

Leu?Glu?Arg?Thr

1

<210>287

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>287

Leu?Arg?Glu?Thr

1

<210>288

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is a nor-leucine.

<400>288

Xaa?Arg?Asp?Ser

1

<210>289

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is a nor-leucine.

<400>289

Xaa?Asp?Arg?Ser

1

<210>290

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is a nor-leucine.

<400>290

Xaa?Glu?Arg?Ser

1

<210>291

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is a nor-leucine.

<400>291

Xaa?Arg?Glu?Ser

1

<210>292

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>292

Lys?Arg?Asp?Ser

1

<210>293

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>293

Lys?Arg?Asp?Thr

1

<210>294

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>294

Lys?Glu?Arg?Ser

1

<210>295

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>295

Lys?Glu?Arg?Thr

1

<210>296

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>296

Lys?Asp?Arg?Ser

1

<210>297

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>297

Lys?Asp?Arg?Thr

1

<210>298

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>298

Lys?Arg?Glu?Ser

1

<210>299

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>299

Lys?Arg?Glu?Thr

1

<210>300

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>300

Lys?Glu?Arg?Leu

1

<210>301

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>301

Lys?Arg?Glu?Leu

1

<210>302

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>302

Lys?Arg?Asp?Thr

1

<210>303

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>303

Lys?Glu?Arg?Ser

1

<210>304

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>304

Lys?Glu?Arg?Thr

1

<210>305

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>305

Lys?Asp?Arg?Ser

1

<210>306

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>306

Lys?Asp?Arg?Thr

1

<210>307

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>307

Lys?Arg?Glu?Ser

1

<210>308

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>308

Lys?Arg?Glu?Thr

1

<210>309

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>309

Lys?Glu?Arg?Leu

1

<210>310

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>310

Lys?Arg?Asp?Ser

1

<210>311

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>311

Lys?Arg?Asp?Thr

1

<210>312

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>312

Lys?Glu?Arg?Ser

1

<210>313

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>313

Lys?Glu?Arg?Thr

1

<210>314

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>314

Lys?Asp?Arg?Ser

1

<210>315

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>315

Lys?Asp?Arg?Thr

1

<210>316

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>316

Lys?Arg?Glu?Ser

1

<210>317

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>317

Lys?Arg?Glu?Thr

1

<210>318

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>318

Lys?Glu?Arg?Leu

1

<210>319

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>319

Xaa?Arg?Glu?Ser

1

<210>320

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine

<400>320

Xaa?Glu?Arg?Ser

1

<210>321

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>321

Xaa?Arg?Asp?Ser

1

<210>322

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>322

Xaa?Asp?Arg?Ser

1

<210>323

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>323

Xaa?Asp?Arg?Thr

1

<210>324

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>324

Xaa?Arg?Asp?Thr

1

<210>325

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>325

Xaa?Glu?Arg?Thr

1

<210>326

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(1)..(1)

＜223〉Xaa is an ornithine.

<400>326

Xaa?Arg?Glu?Thr

1

<210>327

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>327

Trp?Asp?Arg?Ile

1

<210>328

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>328

Trp?Arg?Glu?Ile

1

<210>329

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>329

Trp?Glu?Arg?Ile

1

<210>330

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>330

Trp?Asp?Arg?Leu

1

<210>331

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>331

Trp?Arg?Glu?Leu

1

<210>332

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>332

Trp?Glu?Arg?Leu

1

<210>333

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>333

Phe?Asp?Arg?Ile

1

<210>334

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>334

Phe?Arg?Glu?Ile

1

<210>335

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>335

Phe?Glu?Arg?Ile

1

<210>336

<211>4

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>336

Phe?Asp?Arg?Leu

1

<210>337

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>337

Phe?Arg?Glu?Leu

1

<210>338

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>338

Phe?Glu?Arg?Leu

1

<210>339

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>339

Trp?Arg?Asp?Phe

1

<210>340

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>340

Trp?Arg?Glu?Phe

1

<210>341

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>341

Trp?Glu?Arg?Phe

1

<210>342

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>342

Trp?Asp?Arg?Tyr

1

<210>343

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>343

Trp?Arg?Glu?Tyr

1

<210>344

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>344

Trp?Glu?Arg?Tyr

1

<210>345

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>345

Trp?Arg?Asp?Thr

1

<210>346

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>346

Trp?Asp?Arg?Thr

1

<210>347

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>347

Trp?Arg?Glu?Thr

1

<210>348

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>348

Trp?Glu?Arg?Thr

1

<210>349

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>349

Phe?Arg?Asp?Xaa

1

<210>350

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>350

Phe?Arg?Glu?Xaa

1

<210>351

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>351

Phe?Lys?Asp?Leu

1

<210>352

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>352

Phe?Asp?Lys?Leu

1

<210>353

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>353

Phe?Lys?Glu?Leu

1

<210>354

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>354

Phe?Glu?Lys?Leu

1

<210>355

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>355

Phe?Lys?Asp?Ile

1

<210>356

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>356

Phe?Asp?Lys?Ile

1

<210>357

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>357

Phe?Lys?Glu?Ile

1

<210>358

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>358

Phe?Glu?Lys?Ile

1

<210>359

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>359

Phe?Lys?Asp?Xaa

1

<210>360

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>360

Phe?Asp?Lys?Xaa

1

<210>361

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>361

Phe?Lys?Glu?Xaa

1

<210>362

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>362

Phe?Glu?Lys?Xaa

1

<210>363

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>363

Phe?His?Asp?Leu

1

<210>364

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>364

Phe?Asp?His?Leu

1

<210>365

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>365

Phe?His?Glu?Leu

1

<210>366

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>366

Phe?Glu?His?Leu

1

<210>367

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>367

Phe?His?Asp?Ile

1

<210>368

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>368

Phe?Asp?His?Ile

1

<210>369

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>369

Phe?His?Glu?Ile

1

<210>370

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>370

Phe?Glu?His?Ile

1

<210>371

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>371

Phe?His?Asp?Xaa

1

<210>372

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>372

Phe?Asp?His?Xaa

1

<210>373

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>373

Phe?His?Glu?Xaa

1

<210>374

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<220>

<221>misc_feature

<222>(4)..(4)

＜223〉Xaa is a nor-leucine.

<400>374

Phe?Glu?His?Xaa

1

<210>375

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>375

Lys?Lys?Asp?Ser

1

<210>376

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>376

Lys?Asp?Lys?Ser

1

<210>377

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>377

Lys?Lys?Glu?Ser

1

<210>378

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>378

Lys?Glu?Lys?Ser

1

<210>379

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>379

Lys?His?Asp?Ser

1

<210>380

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>380

Lys?Asp?His?Ser

1

<210>381

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>381

Lys?His?Glu?Ser

1

<210>382

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>382

Lys?Glu?His?Ser

1

<210>383

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>383

Lys?Leu?Arg?Ser

1

<210>384

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>384

Lys?Arg?Leu?Ser

1

<210>385

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>385

Lys?Leu?Arg?Thr

1

<210>386

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>386

Lys?Arg?Leu?Thr

1

<210>387

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>387

Lys?Glu?Leu?Ser

1

<210>388

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>388

Lys?Leu?Glu?Ser

1

<210>389

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>389

Lys?Glu?Leu?Thr

1

<210>390

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>390

Lys?Leu?Glu?Thr

1

<210>391

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>391

Lys?Leu?Arg?Ser

1

<210>392

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>392

Lys?Leu?Arg?Thr

1

<210>393

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>393

Lys?Glu?Leu?Ser

1

<210>394

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>394

Lys?Glu?Leu?Thr

1

<210>395

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>395

Lys?Glu?Ile?Thr

1

<210>396

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>396

Lys?Leu?Arg?Ser

1

<210>397

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>397

Lys?Leu?Arg?Thr

1

<210>398

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>398

Lys?Glu?Leu?Ser

1

<210>399

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>399

Lys?Glu?Leu?Thr

1

<210>400

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>400

Lys?Leu?Arg?Ser

1

<210>401

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>401

Lys?Arg?Phe?Thr

1

<210>402

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>402

Lys?Leu?Arg?Thr

1

<210>403

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>403

Lys?Glu?Ile?Thr

1

<210>404

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>404

Lys?Glu?Val?Thr

1

<210>405

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>405

Lys?Glu?Ala?Thr

1

<210>406

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>406

Lys?Glu?Gly?Thr

1

<210>407

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>407

Lys?Glu?Leu?Ser

1

<210>408

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>408

Lys?Glu?Leu?Thr

1

<210>409

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>409

Lys?Arg?Trp?Tyr

1

<210>410

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>410

Lys?Trp?Arg?Tyr

1

<210>411

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>411

Lys?Arg?Tyr?Trp

1

<210>412

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>412

Lys?Tyr?Arg?Trp

1

<210>413

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>413

Lys?Arg?Tyr?Trp?Thr

1 5

<210>414

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>414

Lys?Arg?Tyr?Thr

1

<210>415

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>415

Lys?Arg?Trp?Thr

1

<210>416

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>416

Lys?Arg?Trp?Tyr

1

<210>417

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>417

Lys?Arg?Tyr?Trp

1

<210>418

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>418

Lys?Arg?Tyr?Trp?Thr

15

<210>419

<211>4

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>419

Lys?Arg?Tyr?Thr

1

<210>420

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>420

Lys?Arg?Trp?Thr

1

<210>421

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>421

Lys?Arg?Trp?Tyr

1

<210>422

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>422

Lys?Arg?Tyr?Trp

1

<210>423

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>423

Lys?Arg?Tyr?Trp?Thr

1 5

<210>424

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>424

Lys?Arg?Tyr?Thr

1

<210>425

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>425

Lys?Arg?Trp?Thr

1

<210>426

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>426

Glu?Lys?Arg?Tyr

1

<210>427

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>427

Lys?Arg?Trp?Tyr

1

<210>428

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>428

Lys?Arg?Tyr?Trp

1

<210>429

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>429

Lys?Arg?Tyr?Trp?Thr

1 5

<210>430

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>430

Lys?Arg?Tyr?Thr

1

<210>431

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>431

Lys?Arg?Phe?Thr

1

<210>432

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>432

Lys?Arg?Trp?Thr

1

<210>433

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>433

Lys?Phe?Trp?Phe?Ser

1 5

<210>434

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>434

Lys?Phe?Trp?Phe?Thr

1 5

<210>435

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>435

Lys?Phe?Tyr?Phe?Ser

1 5

<210>436

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>436

Lys?Phe?Tyr?Phe?Thr

1 5

<210>437

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>437

Lys?Phe?His?Phe?Ser

1 5

<210>438

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>438

Lys?Phe?His?Phe?Thr

1 5

<210>439

<211>6

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>439

Lys?Val?Phe?Phe?Tyr?Ser

1 5

<210>440

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>440

Lys?Phe?Trp?Phe?Ser

1 5

<210>441

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>441

Lys?Phe?Trp?Phe?Thr

1 5

<210>442

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>442

Lys?Phe?Tyr?Phe?Ser

1 5

<210>443

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>443

Lys?Phe?Tyr?Phe?Thr

1 5

<210>444

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>444

Lys?Phe?His?Phe?Ser

1 5

<210>445

<211>5

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>445

Lys?Phe?His?Phe?Thr

1 5

<210>446

<211>5

<212>PRT

＜213〉artificial

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>446

Leu?Phe?Trp?Phe?Thr

1 5

<210>447

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>447

Leu?Phe?Trp?Phe?Ser

1 5

<210>448

<211>7

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>448

Gly?Gly?Gly?Gly?Ser?Ser?Ser

1 5

<210>449

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>449

Lys?Arg?Asp?Ser

1

<210>450

<211>4

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>450

Lys?Arg?Asp?Ser

1

<210>451

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>451

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>452

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>452

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>453

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>453

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>454

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>454

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>455

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>455

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>456

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>456

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>457

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>457

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>458

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>458

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>459

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>459

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>460

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>460

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>461

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>461

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>462

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>462

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>463

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>463

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>464

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>464

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

<210>465

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉peptide of chemosynthesis.Aminoacid can be protection or unprotected D or L type.

<400>465

Asp?Trp?Phe?Lys?Ala?Phe?Tyr?Asp?Lys?Val?Ala?Glu?Lys?Phe?Lys?Glu

1 5 10 15

Ala?Phe

Claims (17)

1. peptide that improves one or more symptoms of struvite situation, the aminoacid sequence of wherein said peptide is made up of Phe-Arg-Glu-Leu shown in the SEQ ID NO:250.

2. the peptide of claim 1, wherein said peptide bag all is made up of D-aminoacid.

3. the peptide of claim 1, wherein said peptide all is made up of L-aminoacid.

4. the peptide of one of claim 1-3, wherein said peptide has the aminoterminal blocking group.

5. the peptide of one of claim 1-3, wherein said peptide has the c-terminus blocking group.

6. the peptide of one of claim 1-3, wherein said peptide has aminoterminal blocking group and c-terminus blocking group.

7. the peptide of one of claim 4-6; wherein said aminoterminal blocking group and/or c-terminus blocking group are independently selected from: tertbutyloxycarbonyl (Boc); Fmoc; nicotinoyl; OtBu; benzoyl; acetyl group (Ac); carbobenzoxy group; methyl ester; ethyl ester; propyl ester; butyl ester; pentyl ester; own ester; N-methylamino o-tolyl; 3 to 20 carbon alkyl; amide; 9-fluorenes Acetyl Groups; 1-fluorenes carboxylic group; 9-fluorenes carboxylic group; 9-Fluorenone-1-carboxylic group; xanthyl (Xan); trityl (Trt); 4-methyl trityl (Mtt); 4-methoxyl group trityl (Mmt); 4-methoxyl group-2; 3; 6-trimethyl-benzenesulfonyl (Mtr); mesitylene-2-sulfonyl (Mts); 4; 4-dimethoxy benzhydryl (Mbh); tosyl (Tos); 2; 2; 5; 7; 8-pentamethyl chromane-6-sulfonyl (Pmc); 4-methylbenzyl (MeBzl); 4-methoxybenzyl (MeOBzl); benzyloxy (BzlO); benzyl (Bzl); 3-nitro-2-pyridine sulfinyl (Npys); 1-(4; 4-dimethyl-2; 6-dioxy ring caproic subunit) ethyl (Dde); 2; 6-dichloro-benzenes methyl (2; 6-DiCl-Bzl); 2-chlorine benzyloxycarbonyl (2-Cl-Z); 2-bromo-benzyloxy-carbonyl (2-Br-Z); benzyloxymethyl (Bom); cyclohexyloxy (cHxO); tert-butoxy methyl (Bum); the tert-butyl group (tBu); trifluoroacetyl group (TFA); 4-[N-{1-(4; 4-dimethyl-2; 6-dioxy ring caproic subunit)-3-methyl dibutyl)-amino } benzyl esters (ODmab); α-allyl ester (OAll); 2-propyloxy phenyl base ester (2-PhiPr); and 1-[4; 4-dimethyl-2,6-dioxy hexamethylene-1-base-subunit) ethyl (Dde).

8. the peptide of claim 7, described peptide is made up of Boc-Phe-Arg-Glu-Leu-OtBu.

9. each peptide of claim 1-8, wherein said peptide and pharmacology go up acceptable mixed with excipients.

10. the peptide of claim 9, wherein said peptide be suitable for going up acceptable mixed with excipients to the Orally administered pharmacology of mammal.

11. the peptide of claim 9, wherein said peptide provides as the unit formulation in the pharmaceutically acceptable excipient.

12. the peptide of claim 9, wherein said peptide provides as time release formulation.

13. being configured to, the peptide of claim 9, wherein said peptide be used for using by being selected from by Orally administered, suction, rectal administration, peritoneal injection, intravascular injection, subcutaneous injection, the approach of using the group that constitutes with intramuscular injection of using, suck of wearing epidermis.

14. the peptide of one of claim 1-13, wherein said peptide and inhibin are co-administered.

15. the peptide of claim 14, wherein said inhibin is selected from the group that is made of cerivastatin, atorvastatin, simvastatin, pravastatin, fluvastatin, lovastatin, rosuvastatin and Pitavastatin.

16. each peptide of claim 1-13 is used for alleviating purposes in the medicine of atherosclerotic one or more symptoms mammal in preparation.

17. each peptide of claim 1-13 is used for the purposes of medicine of one or more symptoms of amelioration of inflammation in preparation.

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