WO2007146823A2 - Carbohydrate derivatives of heptanoic acids - Google Patents

Abstract

In one embodiment, the present invention relates to a compound having the formula (1), wherein R is selected from the group consisting of heterocyclic, aromatic, and cyclic compounds; wherein the dashed line in Formula (1) is selected from the group consisting of single and/or double carbon-carbon bonds; and wherein at least one of R' or R' is a carbohydrate.

Description

CARBOH YPRATE DERIVATIVES OF IIEFΓANOIC ACIDS

INVENTORS:

Anatolc Kiyosov, Ph.D. (36 Walsh Road., Newton. MA 02459) David Plait, Ph.D. 112 Appleion Circle, Newton, MA 02459)

DOCKIiT NO:

089918.024002

US. Paient Cooperation Treaty Application

BACKGROUND OF THE INVENTION

^"This patent application claims priority of Provisional Patent Application Ser, No. 60/804,242 filed June 8, 2006 and entitled "Carbohydrate-Derived Statins".

Cholesterol is a highly decorated small molecule. Thirteen Nobel Frizes have been awarded to scientists who have worked with cholesterol. There are two principal ways for cholesterol to get to the plasma: one is with the diet, another is via biosynthesis. Diet typically contributes iφ to 40% of cholesterol, that enters through the intestine. The

remaining 60% of cholesterol is synthesized from acetyl-CoA via a scries of enzymatic reactions. Eight of these enzymatic reactions are shown below, and the enclosed figure denotes the entire cholesterol biosynthelie pathway.

Λeetyl-CoA ~> Aceioaeetyl-CoA -* β~Hydroxy-β-Methy3giutaryl-CoA -> Mevalonic

acid ™> Geranyl pyrophosphate ~> Famesyl pyrophosphate -> Sqαalεne -> Lanosteroi

-> Cholesterol

■'v'ϊlv ^.v^

\ϊc^ K\

AV-. <* -^^" ^vi

M« valorise ^ckl

O X ^s Λ ϊ

i v> >, \ - Λ ^VV V - -^■>!^■-

i ^'- ^•

CO^ f

V"

G*raπyl PP

s5KN^>M^>M1fS

« C«l;

2,3 «._!κkio$qμ^^n6

f

Xs' ϊ \V λ\ vv. V * VΛ ^V \V. v-

In thi* chain of reactions, a comcrsion of HMG-CoΛ ω mcv alooate is d rate- limuing step. v,hich is catalyzed b\ HMG-CoΛ reductase.

The os frail cholesterol level in plasma ss tight!) controUcd b} a feedback mechanism, the principal element of which i^ that cholesterol is an inhibitor of HMG- CoΛ reductase. An eJexated level of cholesterol leads to the inhibition of the Λifiversion of HMG-CoA to mevalonate. This leads to an increase in HMG-CoA concentration; decreased biosynthesis of cholesterol; and a drop in mevalonic acid concentration, causing deprivation of mevalonatε in the cells. This In turn triggers the accumulation of large amounts of IiMG-CoA reductase (still being inhibited by an excess of cholesterol) in the cell, ami ••■• as a result of said tightly coordinated regulatory events - axs increase of

low-density lipoprotein (LDl,) receptors on the surface of the liver and therefore increased removal of LDL cholesterol from the blood. As a result, the total plasma cholesterol decreases, and the cholesterol balance in the blood, or cholesterol homeostasis, is maintained, even in situations of an occasional excessive absorption of

cholesterol with the diet.

However, in some cases, this feedback mechanism is unable to keep the

cholesterol level in check. This typically results hi hyperiipkiemia, or hypercholesterolemia (high blood cholesterol) due to chronic elevated LDL when the levels of cholesterol hi the blood are too high, such as 6.5 mmol/L (2,5 g/L) or higher.

To correct this situation a certain class of drugs was developed, with a collective name of statins. Statins are a subset of the modified heptanoic acid family of compounds. Statins competitively inhibit HMG-CoA reductase, slow down cholesterol biosynthesis

via competitively inhibiting this enzymatic pathway, and thus increase the level oi^" plasma II MG-CoA with a concomitant increase in LDL receptors on the surface of liver cells and removal of LDL cholesterol from the blood, As a result, total plasma cholesterol decreases, as well as the risk of atherosclerosis and coronary diseases.

4 When a statin Ls added to the system, ii competes with HMG-CoA reductase by interacting with the enzyme binding site. In fact, the β,δ~dihydføxy~heptanøic acid moiety of all statins (in their open acid form) closely resembles the 3-methyl-3-giutaryf binding

moiety of the IiMG-CoA molecule, and this is apparently a primary, functional part of the statins. Most statins also possess a "secondary", hydrophobic function, which

increases their binding to the active site of the enzyme by about 10,000 times.

There are only two chemical functions that belong to all eight statin examples considered here (mevasiatin. lovastatin, pravastatin., simvastatin, iluvastatln, eerivasiatin, atorvastatrø and rosυvastatiπ) and located in the same position of the open acid form of

the statins. They are β,d-diMdroxy groups in the heptanoic (or heptenoic) chain, and the heptanoic (or heptenoic) carboxyite groups. The cycled, lactone form of the heptanoic acid moiety is biologically inactive (or much less active compared to the open acid form), and has to be enzymatieally hydrolyzed to generate the active form. All other chemical groups in statins seem to be replaceable with almost unrelated structures, though hydrophobic ones may be particularly suited to binding with the HM⁽J-CoA binding ϊϊioiety. increase of hydrøphobieity. though within certain stearieal boundaries, seems io be beneficial for competitive inhibition of statins with respect to HMG-CoA reductase.

There are five principal structural elements in statins: (1) the 3^!- and 5 "-hydroxy! groups in the open acid form of the lactone part (β- hydroxy-δ-taetone} of all the statins, or the ^'-hydroxy! group in ihe closed lactone part of statins of microbial origin (mevastatin and iovastatin) and ihe semisynthetic statin

(simvastatin);

(2) ihe side chain bridging ihe lactone (closed or open) and the hydrophobic rnsdeous, that is, the 3.5~dihkkoxy-heptanoic acid moiety of all the microbial statins {mevastatin, iov&st&iin and pravastatins, in the semisynthetic statin (simvastatin) &nd only one fully synthetic statin (atorvastatin), or 3,5-dihidroxy-6-heptenoic acid moiety in three fully syrsihetic staύns (iluvastatin, ceπvastatin and rosuvastatin);

{3 ] ihe carboxylic group of the open lactone raoiety, that is of the heptanoic or heptcnoic acid;

(4) the hydrophobic nucleous moiety — hexahydro naphthalene in ail the microbial and semisynthetic statins (mevastatin, iovasiatin, pravastatin and simvastatin), and substituted heterocyclic nitroaromatic compounds in all fully synthetic statins ffiuvastaiin, cerivastatin, atorvastatin and rosuvastatin); and

(5) the side chain ester moiety, such as ihe 2~melhylbuiyrate ester in all three microbial statins (mevastatin, lovastatin and pravastatin) and 2-dimeihyIbuϊyrate ester in the semisynthetic statin (simvastatin).

6 Statins are a group of compounds generally classified as {a) microbial (for example, raevastatin, iovastatin, and pravastatin); (b) semisynthetic (for example, simvastatin); and (c) fully synthetic (for example, flisvastatin, atorvastatln, and serivastatm). Examples of several natural and synthetic statins are provided below:

O

O

O

O

HzC

CH₃

H^!><< o^X^ H3C^!

Mevastatiπ Lovastatin

HO V /^v ^■Cθ2Na

O O

\/ OH

..^

O ,_'"^" ^ '^■! O

H3C CH₃? H3C ≡ H j-'

^■v, -^v ^CH₃ CH₃

V^

H3C'"^' HO¹

Simvastatin Pravastatin

^ 9^H3

W ^ -H y v

CH3

Fiuvastatin N

'erivastatin Rosuvastatin

Aton astatin The first statin (mevastatin) was isolated in the 1970s. Its formula was identified as f ] S-π α(R*),?βJβ(2S^:5t,4S*}_!8aβ]]-2-methy!bιιtanoic acid l ,2,3,7,8a-hexahydro-7- niethyi~8-[2-(ietøhydro-4-hydroxy-6-oxo-21-ϊ-pyran-2-yl_p)ethyl]-l-naphthaienyl ester, and later was called mevastatin, since it is a substituted meviπic acid lactone (that is tctrahydro-4-hydroxy-6-oxo-2H-ρyran-2-yl).

By 1976 a similar compound, lovasiatin, was isolated from Aspergillus ϊerreus. Instead of C3-hydroge& in mevastatin, iovastatin has a C7-methy! group. The rest of the ϊΩoiecule was the same. The chemical name of lovastatin is [ iS-[1α(R*),3«Jβ,8β (2S*_I4S*),8aβjj-2-methyli>utanoic acid L23;7_!8_>8a~hεxahydR>3,?-dimethyl-8~[2- {teirahydro~4-h_]ydroxy~6-oxct~2ii-p>'ran-2-yl)ethyll-l-naphthak¹siyl ester-

Both statins serve as potent competitive, inhibitors of MMG-CoA reductase, with the inhibition constants close to 1 πM (in tiheir open, water-soluble, acid form, which can be obtained by an alkaline treatment of the lactone form). Since ihe Micliaeiis constant for the natural substrate, HMG-CoA, is close to 10 μfvl mevastatin and lovastatin bind to HMG-CoA reductase 10,000 limes stronger than the natural substrate. Hence, even at a 10QG~tim.es lower concentration compared to that for HMG-CoA, at, 0.1-10 mM of the latter, the statins are able to competitively suppress the enzyme activity and, hence, decrease the conversion of MMG-CoA to mevalonic acid aid then tα cholesterol by 90%.

10 The finding that mevastatin and lovastatin effectively block the biosynthesis of cholesterol that causes the accumulation of HMG-CoA and the respective increase in LDL receptors and the overall clearance of "bat!" cholesterol m plasma, which in turn leads to a lower risk of atherosclerosis and coronary events and the concomitant deaths,

led to a search for more statins. Continuation of research into microbial production of

statins has resulted in one more microbial "fungal statin", named pravastatin. Its chemical

structure was close to those of mevastatin and lovastaim, also of a microbial origin, but the C3-hydrogen in mevastatiri or C3-røethy! group in Jovasiatin was replaced with a C3~ hydroxy group in pravastatin. In addition, a lactone ring in mevastatiπ and lovastatin was substituted with its open form. The chemical name of pravastatin (sodium sail) is [I S- I lαf βS*,δS* ),2α_sόα,8β-(R*),8aαIl- i ;2_s6_>7,8_?8a-heχahydπ>p,δ₅6-trihydroxy-2-methy [-8-

(2-raetliyl~ ! -oxobtUoxy)- 1 -naphthalεneheptanoic acid mcmosodium salt.

Pravastatin has about the same competitive inhibition constant as those for mevastatin and lovastatin, and showed a reduced risk of muscle toxicity, as compared

with iovastatin.

Structural-activity relationship studies have revealed thai the inhibitory activity of the statins in their open, acid form is abolished by the conversion of either of the two hydroxyl-groups φ~ and §-) in the heptanoic chain, linked to the hexahydro naphthalene nueleous, into the methyl tester. Replacement of the carboxyl group of the acid form with carboxaroide also ablated the inhibitory activity of the statins.

I l By chemically modifying iovastatin, a semisynthetic statin, simvastatin, is obtained. It is almost identical with Iovastatin and has the same 3,7-dmietbyl function in the hexahydronapfathalεne ring, except its 1-butanoic acid chain, linked to the hexahydro naphthalene nucleoids, was converted from 2-methylhutaπok acid into 2- dimethyl bmanoic acid. The overall chemical name for simvastatin is [1 S-[Ia Jα,7β,Sp (2S*_>4S*),8aβ]]-2,-2-dimethyIbutanoic acid 1 ,2,3,7 ASa-hexahydro-3, 7-diroeihyI-8-[2- (teti'aiiyclro~4-hydroxy~6-oxo-2II-pyran-2-yl)ethyl]~l -naphthalcn>'l ester.

This additional CHj-group in the butanαic acid side chain has led to doubling to quadrupling in the inhibitory potency of simvastatin compared to pravastatin.

Finally, the fully synthetic statins were made, for example, fluvastatin, ccrivastadn, atorvastatin and rosuvastatin. These fully synthetic statins have a πucleous completely different from the hexaliydro naphthalene nucleoli found in statins from natural sources. Tilt: natural hexahydro naphthalene ring is formed biosyntheiically from acetate units linked to each other in head-to-tail fashion. The active site precursor of the

microbial (and semisynthetic statins on that matter) appears to be the heptaπoic chain cycled to a lactone. The lactone ring in biologically transformed into active metabolites, having an open, acid form of a ! IMG-CoA specific inhibitor. The rest of the active moiecuie serves a tight binding function with respect to HMG-CoA reductase. Hence, the idea of fully synthetic statins was to replace the hexahydro naphthalene nucleus with some other hydrophobic structures, carrying side chains similar to those in the microbial

statins but not necessarily mirroring them.

12 For example, lour fully synthetic statins possess - instead of the hexahydro naphthalene nudeous - Indole (iluvastatin), pyridine (eerivastatln), pyrrole (atorvastaim),

and pyrimidme (rosuvastaiin) nuclei.

The respective chemical names of the fully synthetic statin, examples disclosed

herein are;

Fluvastalm - [R*,S*-(EΪJ ~{±)- 7-[3-{4-πuorophenyi)-I-(1-methylethyi)-lH-

Indol-2-yI]-3,5-dϊhidroxy-6-heptenoic acid,

Ccrivastalin -^■ [R*,S*-( K)J -7-| 4-( 4-tluorophenyi)-5-meihoxymeihyIV 2,6~bJs{ I-methylethy!) 3-ρsτidiny3-3₅5-dihydroxy-6-heptenoic acid.

Atorvastatm - [R-(R*,R*)]-2-(4-fluorophenyl)-β,δ-dihϊdroxy-5-{ ]-inethyiethyl)-

3-phenyI-4-[(pheny lamiπoVcarbonyl^'J- Ui-pysτole~ I -heptanoic acid.

Rosuvas&itisi -bis[(E)-7-[4-(4-flυorophenyl)~6~isopropyI-2-

[methylfmethylsulfoαyl)amino| pyrimidin-5-ylj(3R,5S)-3,5-<iihydroxy-6-heptrøoic acid],

calcium salt.

13 Three out of the four folly synthetic statin examples disclosed here - flisvastatin, eerivastatin and rosuvastatin - contain a similar chemical function, that is an unsaturated

p,δ-dihidroxy-heplenoic acid. Only atorvastaiin contains a saturated, heptanoic acid.

Structural changes in the statins also affect solubility of the statins in water, as well as other physkochemica) properties. Hiss in turn influences the above biomedical properties. All statins are poorly soluble in water, except their acid form at pi I values

above their pK.

For example, iøvasiatm, having a closed lactone structure, dissolves in water at 0.4 μg/mL (it dissolves much better in organic solvents, such as in acetone [47 mg/mLj. ethanoi [16 mg/mL], methanol [28 mg/niLj), Atorvastatin in its sodium salt form (pK 4.46), dissolves in water at pH 2.1 in the concentration of 20,4 μg/mL, but at pH 6,0 it dissolves in the concentration of 1 .23 mg/mL,. Pravastatin in its sodium form is soluble in methanol and water at more than 300 mg/mL.

UMG-COA binds to the MMG-CoA reductase enzyme with the Miehaεlis constant of approximately 2xiø^~sM. in a simplified manner, the Miεhaelis constant is close to a dissociation constant of the enzyme-substrate complex. If a biological concentration of

HMCi-CoA is around 10^'4 M, about 80% of the enzyme is bound with the substrate, After the enzyme-substrate complex (HMG-CoA + HMG-CoA reductase) is formed, the coenzyme, NADPI !, binds to the complex, making it a triple complex of enzyme-

14 substrate-coenzyme. Within this triple complex NADPI ! is oxidized to NADP-CoA, and

HMG-CoA is reduced to mevalonic acid.

The binding of the statins to HMG-CoA reductase proceeds in two steps, ^n the first, initial step, a "simple" interaction occurs, with competitive inhibition constants around 1 nM. In the second step the enzyme-substrate complex undergoes a slow (minutes) conformational change, which increases the inhibition constant by an

additional 20-50 limes, making it into the fractions of the nanomolar range. In the presence of 0.1 mM of HlViG-CoA (5xfC,_T,, that is about 80% saturation of .he enzyme), the "initial" inhibition constants for atorvastatin and rosuvastatin examples were 130 and ! 60 nM, respectively, and after the conformational change of the enzyme, the "steady- state^"' inhibition constants were equal to 44 nM (pravastatin), 28 xiM (ϊluvastatin), 1 1 nM (simvastatin), !0 nM (cerivastatin), 8 nM (atorvastatin} and 3.5 nM (rosuvastatin).

For a comparison, IC₅,; values (which for competitive inhibition depend on the amount of a substrate employed, and for K_m ^::;; 2x10^"" M and the amount of HMG-CoA of Hf⁺M would be 6 limes higher than the respective K, values) for the statins were; 28 nM (fiuvastatin). 23 nM (mevaslatin). I I nM (simvastatin), IO nM (cerivastatin}, S nM (atorvastatin}, and 5 nM (rosuvastatin).

Previous data have shown that a I C ₅₀ value for the ML-236B compound (later named as mevastaun) was equal to 0.01 μg/mL. Since the molecular weight of mevastatin is 1000 Da., this !C_So is actually equal to 10 nM, close to the above figures.

15 The above eorαpeiuive inhibition figures show that all the statin examples above demonstrate similar Inhibitory properties.

The bioavailability or amount of statins reaching the systemic circulation is

generally in a 10-30% range, and most statins are tightly bound with plasma proteins. Fo? example, systemic bioavailability of fϊuvasfatin has been reported as 24%, pravastatin

17%, atorvastatin 14% (some data put it at 30%), lovastatln and simvastatin more than 5%. According Io other data, the absolute oral bioavailability of atorvastatin (Lipiior) varies between 12% and 14%, fϊuvastatin 19-29%, pravastatin 18%, iovastaiin and

simvastatin 5%, rosuvastafin 20%

Maximum plasma concentration of the statins is reached at 2-4 hours for lovastatin, 2-3 hoars for atorvastatin and rasuvastatin. 1.3-2,4 hours for simvastatin, 0.9- 1.6 hours for pravastatin, and 0.5-1 hours for liuvastatin.

Maximum plasma concentrations (C_m8x) of the statins - based on a 40-mg oral dose - were reported to be 448 ng/mL for flovastaiin, 45-55 ng/mL for pravastatin, 27-66 ng/niL for atorvastatin, 37 ng/mL for rosuvastatin, and 10-34 ng/ml, for lovastatin and

simvastatin (Ibid).

16 SUMMARY QF THE INVENTION

In one embodiment of the claimed invention, a compound is disclosed having the formula (I), wherein R is selected from the group consisting of heterocyclic, aromatic,

and cyclic compounds; wherein the dashed Hue in Formula 1 is selected from the group consisting of single or double carbon-carbon bonds; and wherein at least one of R^* or R^"

is a carbohydrate.

R

(Formula ϊ)

In another embodiment, at least one of R' and R" of Formula 1 is a

mono sacc hart de .

in. another embodiment, at least one of R^' and R^"' of Formula l is a galactose .

In another embodiment, R of Formula 1 is selected from the aromatic group

consisting of condensed aromatic, substituted aromatic, hcxahydronaplithalene, and

aromatic derivative compounds. In another embodiment, R of Formula lis selected from a heterocyclic group consisting of substituted heterocyclic, heterocyclic derivative, aromatic heterocyclic, ttitroaromatic, indole, pyrrole, pyridine, and pyrrolidine compounds.

In another embodiment, R of Formula I is selected from the cyclic group consisting of cyclic, substituted cyclic, and cyclic derivative compounds,

In another embodiment, R of Formula 1 is further comprised of a side chain

selected from the group consisting of butanoic acid, 2-methyibutanoic acid, or 2-

dimethyibutanoic acid.

In another embodiment, R of Formula 1 is Formula 4, wherein the dashed line represents a chemical bond between R and Formula 1 :

(Form u Sa 4)

In another embodiment, X of Formula 4 is a monosaccharide. in another embodiment, X of Formula 4 is a galactose,

In another embodiment, the compound of Formula 1 is a lactone.

DETAILED DESCRIPTION OF THE INVENTION

Detailed embodiments of the present invention are disclosed herein; however, it is to be understood that the disclosed embodiments are merely illustrative of the Invention that may be embodied in various forms. In addition, each of the examples given in connection with the various embodiments of the invention are intended to be illustrative,

and are not restrictive, Therefore, specific structural and functional details disclosed herein are not to bε interpreted as limiting, but merely as a representative basis for teaching one skilled in the atl to variously employ the present invention.

The following terms shall have the meanings indicated herein, unless required otherwise by the context.

"Acetyl CoA" refers to a component in cholesterol synthesis. Acetyl Co-A is the precursor to HMG-CoA, which, in animals, is a component in cholesterol and ketone synthesis

"Adrnir&Istra.ioii" refers to oral, or parenteral including intravenous, subcutaneous, topical, transdermal, intradermal, transmucosat intraperitoneal . intramuscular, intracapsular, intraorbital, intracardiac, , transtracheal subcutaneous.

19 subcuticular, intraarticular subcapsular, subarachnoid, intraspinal epidural and/pr inirastemal injection and infusion,

"Ammosugar" refers to a sugar containing an amino group m place of a hydroxy! group in the sugar molecule, and inc!uding,hut not limited to, N-aeeiylglueosamine. The

following structure is an example of an aminosugar:

"Aromatic compound^*' refers to an organic molecule that contains at least one aromatic rings, Suitable aromatic compounds include, such as. but are not limited to, benzene, pyridine, and indole. These organic molecules are typically characterized by a

high degree of saturation, contain alternate single and double bonds when presented in a simplified way, and exhibit resonance between the single and double bond states. The

following structure is an example of an aromatic compound;

20 '-Aromatic derivative compounds" refers to an aromatic compound containing at least one functional group which can be obtained from the initial aromatic compound in one or several .sequential chemical reactions. It is commonly referred to a chemically

substituted aromatic compound.

"* Aromatic heterocyclic compound" refers to an eiemenio-organic compound containing a ring struciure(s) which contains atoms in addition to carbon, such as nitrogen, oxygen or sulfur, as part of the ring. These compounds may be either simple aromatic rings or fused, or condensed aromatic rings, which include, bin are not limited

to, indoles.

"Rutanoic acid" or butyric acid means a carboxylic acid with structural formula i-ϊjsCHs€I-b-COOH. The following structure is an example of hutanoic acid:

'"Carbohydrate" means any molecule consisting of carbon, oxygen and hydrogen

atoms and having a general formula (CHjO)_n. Carbohydrates contain alcohol (aldose) or ketone (ketose) factional groups, and range from the simple sugars, or monosaccharides, to the polysaccharides. "Condensed" or "fused" aromatic compounds refer to a group of organic molecules consisting of two or more fused benzene rings.

"Cyclic compound" refers to an organic compound in which a series of carbon atoms are connected together to form a loop or ring. These compounds do not have alternate single and double chemical bonds. These compounds are alternately referred to

as cyclσalkanεs. They do not contain nitrogen, oxygen or sulfur atoms in the cycle.

"Cyclic derivative" refers to a cyclic organic compound which can be obtained from the initial cyclic compound in one or several sequential chemical reactions. Il is commonly referred to a chemically substituted cyclic compound.

"Decreased total cholesterol" means a decrease in mammalian plasma overall cholesterol level of greater than about 10% as compared to an initial iota! cholesterol teve! of a subject and/or a measurement of mammalian plasma overall cholesterol level of a control mamma! fed an identical diet. Total cholesterol is assayed by heparin-MnCb precipitation, or as assayed by use of polyethylene glycol 6000 precipitation, or using so-

called automatic ^'Triple Lipid Screening Test (for total cholesterol, HD L -cholesterol and triglycerides) and its modifications. These assays can be conducted enzymatieaOy with a cholesterol kit (cholesterol ES-reagent, Beekman Instruments, Inc.) (Mao. S. JX and Kottke, B, A, Determination of high-density lipoprotein cholesterol by heparm-MnCb precipitation. Clin, Chern., v. 26, 1369, 1980} (hereinafter incorporated by reference} or by polyethylene glycol 6000 precipitation (Izzo, C, Grilio, F. and Murador, E. Improved method for determination of high-density- lipoprotein cholesterol. Isolation of high-

?? density lipoproteins by use of polyethylene glycol 6000. CHn. Chem,, v. 27, 371 -374, 1981) {hereinafter incorporated by reference).

^■'Decreased total low-density lipoproteins (LDL)" means a decrease in overall low-density lipoproteins in. plasma of greater than about 10% as compared io an initial overall lipoprotein level of a subject and/or a measurement of mammal ian plasma overall low-density lipoprotein level of a control mammal fed an identical diet. Overall iow- density lipoproteins may be assayed by uliracenirifugatioπ, or by using an imniunoseparation or specific detergents to separate LDL panicles from other

lipoproteins, followed by measurement of cholesterol by conventional enzymatic reactions. This assay may be conducted as taught, in Brian D. Ragland, Robert J. Konraii, Carolyn €haffiπ, C. Andrew Robinson and Robert W. Hardy (Clin, Chem, 24, No, I K 1848-1851 , 2000). which is herein incorporated by reference. LDL-choksiero! in plasma can also be calculated based on total cholesle.ro! minus HDL-ehαiesteroI and

triglycerides.

"Decreased serum osteocalcin" means a decrease m serum osteocalcin (as assayed by radioimnmneassay as m U.S. Patent No. 6,756,401, herein incorporated, by reference) and as compared to serum osteocalcin levels prior to treatment or in an untreated mammal.

^iyDisaeeharkk" means a carbohydrate composed of two monosaccharides.

23 "Effective dosε" refers to a (lose of a compound that improves the symptoms of the subject or the longevity of the subject suffering from or at high risk of suffering from hypercholesterolemia, cancer, or other diseases. The effective dosε hi embodiments of this Invention can. he quantitatively defined as an amount of a polysaecharide-derived statin administered alone or m a mixture with a dose of a therapeutic agent administered in a subject for treating hypercholesterolemia or cancer, or other disease that decreases a level of a chosen symptom by at least 20 per cent, or between 5 and 50 per cent and more,

depending on an advancement of the disease.

"Efficacy" of a therapeutic agent refers Io the relationship between a minimum effective dose and an extent of toxic side effects. Efficacy of an agent is increased If a therapeutic end point can be achieved by administration of a lower dose or a shorter dosage regimen. If toxicity can be decreased, a therapeutic agent can be administered on a longer dosage regimen or even chronically with greater patient compliance and improved quality of life. Further, decreased toxicity of an agent enables the practitioner

to increase the dosage to achieve the therapeutic endpoint sooner, or to achieve a higher

therapeutic endpoint.

^•'Enhance" means to alter the relationship between a given dose of a compound

and its effect on a mammal or mammalian ceil, so that the given dose of a compound will have a greater effect (positive or negative) on the given mammal or mammalian cell than

the given dose of the compound.

24 "Galactose^" refers to a monosaccharide, in which ihe first wά ihe last hvdroxvlgroups point the same \\a> and the second and third livάruxyl groups point ihe o:her wsy. The sullowmg structure it* an example of galactose"

CHPH

O OH OH

OH

C

π i^ *™¹*¹ \ '-^•-^•-^•-^•-'^■Λ ^

H — f' — OH

t ^'HjOH

"i lctCioc>eHe compound" is an elcmonlo-organie compound coutaming a ring struiti.u-c(s ) which contains atoms in addition m earbt>π, such as ratrogeu, o\\gcn or sulfur, as pan of the ring. Heterocyclic cυrnpounds are either ^insple aromatic ring^ or non-arornatic rings, which include but are not limited io pyridine. The following structure is an example of a heterocyclic compound:

Fhe following is the structure of pyrimidine:

N

¹^ .

N

"^kI Ieterocyde derivative compound^"' refers to an organic compound containing at least one heteraeyde which can be obtained from an initial heteroeyele compound in one or seveπil sequential chemical reactions. It is commonly referred to a chemically

substituted heteroeyele compound.

"Hexahvdronaphihaiene compound"¹ refers to a condensed or fused aromatic compound having two or several fused benzene rings, which includes but not is not limned to the following structure:

26 And the following structure:

And the foHowhig structure:

And the following structure:

"TIMCJ - CoA^v HMG-CoA (or 3-hydroxy-3-met%!giutaryi-coens!yme A) means an intermediate in the Mevalonate pathway, formed from acetyl CoA and acetoacety!

CoA by HMG-CoA synthase.

'1-IMG-CoA reductase" means an enzyme that converts HJMG-Co.A to mevalonic

acid.

2? ¹IIMG-CoA reductase inhibitors^'", or "statins", means a class of compounds that lower cholesterol by inhibiting the enzyme FIMG-CoA reductase, which .us the rate-

limiting enzyme of the mevalonate pathway of cholesterol synthesis,

'Ηypoeholesieroiemic effective amount" is an amount of a compound that decreases plasma levels of cholesterol (HDL₅ LDl., or combination.) in the plasma of a

mammal by at least 10%,

'Increase mean plasma elimination half iife" means an. increase of the residence time of a compound in question in the blood (in plasma) until the concentration of the compound reaches 50% of the Initial concentration (or the maximum concentration, or its radioactivity when a radiolabeled compound is used) of the administered compound in plasma. A noticeable increase of the half-life time is when it exceeds the error margin of

the measurement of the half-life time by at least twice,

"indole" means an aromatic heterocyclic organic compound with a hieyclie structure, consisting of a six-membered benzene ring fused to a five-mernbered nitrogen- containing pyrrol y ring. The following structure is an example of an indole:

~~-^ /

28 "Inhibit cholesterol biosytttheses" means in vitro or in vivo inhibition of IiMG- CoA reductase activity as compared io a control. This inhibition is demonstrable using the methodology of Beg et. _<il, FEBS Letters 80: 123 - 129 (1911) or as described in J. Bio). Cheni. 234; 2835 (1959), herein incorporated by reference.

'improved biodistribution" means increased exposure of tissue to a compound, for example through binding of the compound or portion of a compound to rmuranaiian cells and/or increased uptake of the compound and/or portion of a compound by mammalian cells.

"Lactone" refers to a cyclic ester, which is an intramolecular condensation product of an alcohol group and a carbαxylic acid group.

"Mean plasma elimination half life" means the average time required until the concentration of a compound in question in the blood (in plasms) reaches 50% of the initial concentration (or maximum concentration, or its radioactivity when a radiolabeled compound was used) of the administered compound in plasma. It can also bε defined as a lime span until half of the administered compound is eliminated from, (he bloodstream.

"Modified heptanoic acid compound" means a compound (including

pharmaceutical salts and/or any alternative structures) having any of formulas ( I) - (3). "Modified heptanoic acid compound'^* includes as a subset of the group the class of compounds described here as '"statins". The following structures are examples of modified heptanoic acid, compounds:

29 NHfTV sP

HOs s-o ^

,vOR"

$w

> φ' ^p: ..^

\ ^^R' J^ ^ .QK

s&

R R

(Formula 1} (Formula 2) (Formula 3}

"Moiety" means a functional group, or part of a molecule.

"Monosaccharide" refers to polyhydroxyaldehyde (aldose) or polyhdroxyketotie (keiose) and derivatives and analogs thereof.

'"Niiroaromatk compound" refers to an elememo-organic compound conUiining a ring structure(s) which contain at least one nitrogen as part of the ring. There may be either simple aromatic, rings or fused, or condensed aromatic rings, which include but are not limited to indoles.

"Oligosaccharide" refers to a linear or branched chain of monosaccharides that includes up Io about 10 saccharide units linked via glyeosϊdic bonds.

"Patient" shall mean a human subject who has presented at. a clinical setting with a particular symptom or symptoms suggesting the τmed for treatment.

30 "Pharmaceutically acceptable carrier" refers to any and ail solvents, dispersion media, e.g., human albumin or cross-linked gelatin polypeptides, coatings, antibacterial and antifungal agents, isotonic, e,g., sodium chloride or sodium glutamate, and absorption delaying agents, and the S ike that are physiologically compatible. The use of such media and agents for pharmaceutically active substances is well known in the art. Preferably, the earner is suitable for oral, intravenous, intramuscular, subcutaneous. parenteral spinal or epidural administration (e.g., by injection or infusion). Depending on the route of administration, the active compound can be coated in a material to protect the compound from the action of acids and other natural conditions thai can inactivate the

compound,

"Plasma" means the liquid component of blood, in which the blood cells are suspended.

"Polysaccharide" means a linear or branched chain of monosaccharides which

includes about 10 or more saccharide units linked by glycosidic bonds,

"Pyridine" refers to a heterocyclic aromatic compound similar t.o benzene and pyrimidme, containing one nitrogen atom at position 1 in the six-member ring. The

following is an example of the structure of pyridine;

N

31 "Tyriinidme" refers to a heterocyclic aromatic compound similar to benzene and pyridine, containing two nitrogen atoms ai position 1 and 3 in the six-member ring. The following is an example of the structure of pyrarmdine;

^"H

S> N

"Pyrrole" refers to a heterocyclic aromatic organic compound, a five-merøbered ring with the formula €4-1 <N. The .following is an example of the structure of pyrrole:

-NH

"Saccharide" refers to any simple carbohydrate including monosaccharides, monosaccharide derivatives, monosaccharide analogs, sugars, including those, which form the individual units in an oligosaccharide or a polysaccharide.

"Side chain^"" means s part of a molecule that is attached to a core structure. An R group is a generic label for a side chain which can be anything; however, it is typically

stable and covaknϋy linked to the adjoining atom.

32 "Statins" refer Io a group of hypolipidemic agents which possess inhibitory properties with respect to HMG-CoA reductase in the liver (or isolated from the liver), and thereby lower blood cholesterol levels in mammals. The statins include but not limited to Mevastatin, Lovastatin. Pravastatin, Simvastatin. Fiuvastatin, Cerfvasiatiπ,

R o s u vastati n , A torvastat? n .

"Subject" is defined here and in the claims as a mammal including a human in need of therapy for, or susceptible to, a condition or its sequelae. The subject may include dogs, cats, pigs, cows, sheep, goats, horses, rats, and mice and humans; and cells and cellular cultures derived therefrom. The terra "subject" does not exclude an individual

that is normal in all respects,

"Substituted aromatic compound" refer to an aromatic compound covaientiy attached to another chemical group, forming thereby a new chemical entity.

"Substituted cyclic compound^'' refers Io a cyclic compound covaientiy attached to another chemical group, forming thereby a new chemical entity.

""Substituted heterocyclic compound" refers to a heterocyclic compound covaientiy attached to another chemical group, forming thereby a new chemical entity.

33 "Systematic bioavailability'^" means the fraction (in per cent) of the compound reaching the mammalian systemic circulation compared with the administered amount,

measured by any appropriate analytical procedure.

"'Thiosugar^'" refers to a sugar containing either a sulfur atom in the cyclical group of the sugar or & sulfur-containing functional group attached to the sugar molecule, including but not limited to 5-thiomethyl xylose. Tiie following is an example of a

thiosugar:

HO

HQ

"Treatment" may be applied to the reduction or prevention of abnormal cell proliferation, cell aggregation and cell dispersal (metastasis).

In one embodiment of the invention, a compound is disclosed having the formula (1), wherein R is selected trom the group consisting of heterocyclic, aromatic, and cyclic compounds; wherein the dashed ime in Formula I is selected from the group consisting of single or double carbon-carbon bonds; and wherein at least one of R^! or R" is a carbohydrate.

34 HO-

\.<^

_Λv*0R"

\ _,,^^R>

R

(Formula 1}

In another embodiment, at least one of R' and R^s' in Formula 1 is a polysaccharide,

hi another embodiment, at ieast one of R" and R" in Formula ! is an oligosaccharide.

In another embodiment, at Ieast one of R' and R" in Formula 1 is a di saccharide.

hi imoύisτ embodiment, at least one of R' and R" in Formula 1 is a monosaccharide.

In another embodiment, at least one of R.^" and R" in Formula 1 is a simple sugar,

In another embodiment, at least one of R^% and R" in Formula I ss a galactose.

35 In another embodiment R in Formula I is selected from the aromatic group consisting of condensed aromatic, substituted aromatic, hexahydronapbihalene, and aromatic derivative compounds.

In another embodiment R in Formula 1 is selected from the heterocyclic group consisting of substituted heterocyclic, heterocyclic derivative, aromatic heterocyclic, siitroaromatic heterocyclic, indole, pyrrole, pyridine, and pyraniidine compound.

In another embodiment, R in Formula 1 is selected from the cyclic group consisting of cyclic, substituted cyclic, and cyclic derivative compounds.

In another embodiment, R in Formula 1 and as shown in Formula 4 is further

comprised of a 1-butanoic acid side chain.

Ia another embodiment, the 1 -butanoic acid side chain of Formula 4 is 2-

πKtfrylbutanoic acid.

In another embodiment, the 1-butanoic acid side chain of Formula 4 is 2- dimeihylbutanoie acid.

In another embodiment having the formula (2), wherein R is selected from the group consisting of heterocyclic, aromatic, and cyclic compounds; the dashed line in

36 Formula 2 is selected from the group consisting of single or double carbon-carbon bonds; and wherein at least one of R\ R", or R"' is a carbohydrate.

^0R'"\ ^⁰

J^

^m

f

(Formula 2)

In another embodiment, at least one of R'. R" and R'^*' in Formula 2 is a

polysaccharide.

in another embodiment, at least one of R\ R" and R"^* in Formula 2 is an oi igosacchar ide .

IB another embodiment, at least one of R', R" and R"^: in formula 2 is a

disaccharide.

In another embodiment, at least one of R', R" and R'" in Formula 2 is & monosaccharide.

In another embodiment, at least one of R\ R" and R⁵" in Formula 2 is a simple suaar.

37 In another embodiment, at least one of R\ R" and R"^" of Formula 2 is a galactose.

In another embodiment, R in Formula 2 is selected from the aromatic group consisting of condensed aromatic, substituted aromatic, hexahydronaphthalene, and/or

aromatic derivative compounds.

In another embodiment, R in Formula 2 is selected from the heterocyclic group consisting of substituted heterocyclic, heterocyclic derivative, aromatic heterocyclic, nitroaromatic heterocyclic, indole, pyrrole, pyridine, and pyramidine compounds.

hi another embodiment, R in Formula 2 is selected from the cyclic group consisting

of substituted cyclic, and cyclic derivative compounds.

In another embodiment, R in Formula 2 is further comprised of a ! -btuaπoic acid

side chain as shown in Formula 4.

In another embodiment, the 1-bιrtaπoie acid side chain of Formula 2 and shown in

Formula 4 is 2-methylbutanok acid or 2-dimethyibutanoic acid,

In another embodiment, the compound of Formula (3), wherein R is selected from the group consisting of heterocyclic, aromatic, and cyclic compounds; the dashed line in

38 Formula 2 is selected from ihe group consisting of single or double carbon-carbon bonds; and wherein at least one of IV, R", and R'" is a carbohydrate.

ji AOR"

_s\OR-

>*^•"

R

(Formula 3}

In another embodiment, at least one of JV, R" and R"' in Formula 3 is a

polysaccharide.

hi another embodiment, at least one of R\ R" and IV" in Formula 3 is an oli gosace h&ride,

in another embodiment at least one of R', R" and IV^* in Formula 3 is a di saccharide,

IB another embodiment at least one of R.\ R" and R^1-" in Formula 3 is a

monosaccharide.

In another embodiment, at least one of R', R" and IV" in Formula 3 is a simple sugar.

In another embodiment, a.t least one of R', R"' and R"' Ln Formula 3 Is a galactose.

39 In another embodiment, R in Formula 3 is selected from the aromatic group consisting of condensed aromatic, substituted aromatic, hexahydronaphthalenε, and/or

aromatic derivative compounds.

In another embodiment, R in Formula 3 is selected from the heterocyclic group consisting of substituted heterocyclic, aromatic heterocyclic, nitroaromatie heterocyclic,

indole, pyrrole, pyridine, and pyramiding compounds.

In another embodiment, R in Formula 3 is selected from the cyclic group consisting

of substituted cyclic, and cyclic derivative compounds.

ϊn another embodiment, R of Formula 3 and Formula 4 is further comprised of a 1 -

butanoic acid side chain.

Io another embodiment, the 1 -butanoic acid side chain of Formula 3 and shown in

Formula 4 is 2-methylbutanoic acid.

In another embodiment, the 1 -butanoic acid side chain of Formula 3 and Formula 4 is

2-dimethylbutanoie acid.

In another embodiment, R in Formulas 1, 2 and/or 3 is Formula 4, and the dashed

line represents a chemical bond between R and Formula 1, 2 and/or 3: h£.

(Formula 4)

In another embodiment. X ^"m hormula 4 is a carhohvdrate.

in another embodiment. X in Formula 4 is a pol^ysaccharide.

In another embodiment, X in Formula 4 is an oligosaccharide.

hi anothoi embodiment. X in Formula 4 is a disaccharidc.

. another embodiment, X m Kormula 4 is a monosacchaiidv

HJ another embodiment. X m Fcsπnula 4 is a simple sugar.

In another embodiment. X in Formula 4 is a galactose.

ϊn anotlu'r embodiment, the cvjnψoimd shown in Formula 1 , 2. and or 3 exists in

the lactone fomi. In another embodiment, ihe invention, relates to a compound oϊ ihe structure 2- metliyf-birtyπc acid g-[2-{4-hydroxy-6-oxo-tetτahydro-pyran-2-yϊ)-eth>^jlj-7-sϊjethyi-3-(D-

galactopyraiiosyloxy)- 1 ,2,3 JASa-hexahydro-naphthalen-l -yl ester,

In another embodiment, the invention relates to a compound of ihe structure 2- rnethyl-butyrie acid 8-[2-(4-[D-gaiactopyranosyloxy]-6-oxo-tctτahydi-o-pyran-2-yf}-

elhyI]-7-methy]-3-hydroxy- 1 ,2,3₅7,8,8a-hexahydro-ϊia.phtha!en-l -y ^j ester,

In a fltrther embodimeni, the invention relates to a compound of the structure 2- ffiethyl-butyric acid S-f2-(4-[D-galactopyranosyloxy)-6-oxo-tetraiiydro-pyran-2-yl)-

ethyi]-7-methyl-3~(D-ga]actopyranosyloxy)-l,2,3_s7₎8,8a-hexahydro-iiaphthalen-1-yi

ester.

ϊn yet another embodiment, the invention relates to a compound of ihe structure

[l S-[Iϋ(βS*_JδS*),2ϊi,6fi,8β*(R*)_}8aαJJ-l,2,6_J7_>8_t8a-hexahydro-β,δ-<iibydroxy-6-CHβ-ϊ>- galactopyranosyk>xy)-2-methyl-8-(2-raethyI- 1 -oxobutoxyV 1 -naphthalenehepiaiioic acid

fflouosodium salt.

ki another embodiment, the invention relates to a compound of the structure [ IS- [ Jα(βS*_}δS*),2α,6o3[^(R*),8aα]]-K2Λλ8,8a-hexahydτo-β,S-dihydroxy-6^MCH2-CH2- CH_?.-CH₂-β-D-ga!actopyraϊiosyϊoxy)-2-meΛyl-8-{2-methyl-l-oxobutoxy)-ϊ-

naplithalendii'ptanoic acid monosodiurπ salt.

42 In another embodiment [!S-[lα(βS*,δS*},2α_>6α_t8βHR*),8aα]]- j_s2,6,7,8,8a-

hexahydro-β,δ,6-trihydroxy-2-metiiy!-8-(2-methyl-I-oxobutoxy)-l-naphthaieϊieheptanoic

acid β-D-galaεtopyranosylϋxy ester.

Ia another embodiment, the invention relates to a compound of the structure [1 B- [lα(βS*_sδS*),2α,δ«.8β<R*)3aα]3-l,2-6J3,δa-hes^ydro^_>δ,6-trihydroxy-2-raethyI-8-

(2-methy 1- 1 -oxobutoxy )- 1 -naphthaleneheptanoic acid (CHj)._! ^■ β-D-gaϊaclopyranosyloxy

ester,

In another embodiment, the invention relates to a compound of the structure [R- {R*,R*}|-2-(4-1luorophenyi)-β-0-(β-D-galaciopyraiiosyIoxy).δ-hydroxy-5-(l- niethy I etby [}-3 -pheny I -4- [(phen ylamino)carbony!]- 1 H -pyrrole- 1 -heptanoi c acid Iie.mi

calcium.

In another embodiment, the invention relates to a compound of the structure [R- (R*,R*)]-2-(4-f1uorophenyl)-β-hydτOxy^^(β-D-galactoρyranosyioxy)-S-{1 - methylethy [}-3-pheny l-4-[(phenyIammo)carbonyl]~11-i-pyrroIe- 1 -heptanoic acid hemi

caleiurø.

In another embodiment, the invention relates to a compound of the structure [R- (R*.R*)J-2-(4-f1uorophenyI)-β_{δ-dihydioxy-5-(l-methylethyI)-3-ρhenyϊ-4- [(phcnyjamino)carbonyl]- lH-pyrrole-1 -heptanoic acid β-D-galactopyranosyioxy ester.

43 In another embodiment, the invention relates to a compound of the structure [R- (R*,R*)]-2-(4-fluorophenyl>-p,δ-dihydroxy-S-(l-meΛyJcihyϊ)-3-pheny!-4- [{pheny iamino)car bony3|-l I I-pyrrole-1 -heptanoic acid. amide N"-[β~D-

gaS actopyranosy !oxy-{ 1 -^ 6)-O- 1 -deoxy-D-gluert- 1 -y 1] .

In another embodiment, the inveniioπ relates to a compound of the structure [R- {R*,R*)j-2^4-πuoroρhenyl>β_!δ-hydroxy-5^1-inethyleth>'i)-3-phenyl-4- [fpheny!amino)carbonyi]-lH-ρyrrole-1 -beptanoic acid amide N"-[β~D- ga!actoρyranosyioxy-(l ~>6)-0-l-deoxy-D-g!uch-l-yi].

In yet another embodiment the invention relates to employing each of the modified heptanoic acids disclosed and/or claimed herein as hypochoiesterolemic agents.

In another embodiment, the compounds of the present invention inhibit cholesterol biosynthesis in a patient,

In yet another embodiment, the compounds of the present Inveniioπ may be administered as a pharmaceutical composition, alone or in combination with a

pharmaceutically acceptable carrier, such as but not Limited to those as described above.

In another embodiment, the compounds of the present invention may be administered as aα anticancer agent, alone or in combination with a pharmaceutically

44 acceptable carrier, by administering an effective amount of said compound Io a patient in

need of such treatment.

As nseά herein, an "anti -cancer agent" is an agent that inhibits, for example,, the following cancers: chronic leukemia, breast cancer, sarcoma, ovarian carcinoma, rectal

cancer, throat cancer, melanoma, colon cancer, bladder cancer, lung cancer, mammary adenocarcinoma, gastrointestinal cancer, stomach cancer, prostate cancer, pancreatic

cancer, or Kaposi's sarcoma.

In another embodiment, the compound of the present invention may be in a powder form.

In another embodiment, the compound of the present invention may be in a liquid

form.

In another embodiment, the compounds of the present invention comprise an amount of a modified heptanoie acid that reduces a toxic effect in the subject by at least 20%, the toxic effect resulting from administration of a known chemotherapy drug and measurable by the respective grade, by weight loss of the subject., by decrease of rcύ blood ceils, white blood cells, platelets, by increase of other unfavorable biochemical and other indications in the subject, such as hair loss. eic.

45 hi another embodiment, the compounds of the present invention may he administered to enhance the hypochoieslerolerme effect of a corresponding hepianoic

acid,

in another embodiment, an effective dose of the compounds of the present

invention may enhance the decrease of total serum cholesterol

In another embodiment, an effective dose of the compounds of ihe present

invention may enhance the decrease of LDL cholesterol

In another embodiment, an effective dose of the compounds of the present

invention may inhibit cholesterol biosynthesis.

In another embodiment, an effective dose of the compounds of the present invention may inhibit MG-CoA reductase activity.

In another embodiment, an effective dose of the compounds of the present invention may increase mean plasma elimination half life as compared to unmodified heptanoic acid and heptaneie acid derivatives.

in another embodiment, an effective dose of the compounds of the present

invention may improve btodistribution of the compound.

46 In another embodiment, an effective dose of the compounds of the present

invention may suppress serum osteocalcin levels.

The following example of synthesis of the compounds of the present invention is

merely illustrative and do not serve to limit the scope of the Invention.

For example, glycoside derivatives of substituted i -naphthaieneheptanυic acid

(pravastatin, mevaslatin. iovastaun, simvastatin), 7-indoiheptenoie acid (fiuvastatm), 7- pyτidmvihq-jtenoic acid (cerivastatin), i-pyrroteheptanok acid (atorvastathO, 7- pyrimidmheptenok acid (rosiivastatin) may be synthesized by reaction of the respective hydroxy] groups with the respective monosaccharides using the Fischer .method, employing hydrogen chloride as a catalyst. Formation of carboxyltς acid esters of substituted derivatives of 1-naphthaieneheptanok acid. 7-indoiheptenoic acid, 7-

pyridinyfheptenok acid, l-pyrroleheptanoic acid and 7-pyrimidmhepteπoic acid are achieved using an acid catalyst, such as zinc chloride, in a pyridine solution of the

carbohydrate at 24⁰C.

In another example, modified heptanoic acids of substituted 1 - naphihaienehepianoic acid (pravastatin, mevastatin, Iovastatin. simvastatin), 7-

indolheplenoic acid (tluvastatin), 7-pyridinylheptenoic acid (cerivastatin), 1~ pyrroieheptanoie acid (atorvastatϊn), 7-pyrimidinheptenoic acid (rosuvastatin) may be assayed as (a) inhibitors of HMG-CoA reductase in vitro, (b) suppressors ofhuman breast iuraor eel! growth in vitro, (e) suppressors of COLO-205 human colorectal cancer tumors

47 in male N€r-nu atliymic nude mice, (ά) suppressor of estrogen deficiency bone loss in ovaxiectαrruzed rat model in vivo, and/or (e) suppressor of cholesterol levels in New-

Zealand white rabbits model m vivo.

Example I

Modified heptanoie acids of the present invention may be tested as inhibitors of HMCs-CoA reductase enzyme by the method described in U.S. Pat. No. 4,231,938 using enzymes prepared as described in the '938 patent At 0.02 mg/mL the Inhibition is more

than 95%, and IC\y is estimated between 0.7 and 1.5 nig/raL, indicating thai the compounds tested are very potent inhibitors of sterol synthesis acting at the B MG-CoA

reductase level

Example 2

Suppression in vitro of human breast tumor call growth with modified hepianoiε acids of the present Invention may be tested using MCF- 7 eel! line. The cells may be incubated with each of the obtained glycoside derivatives at their various concentrations for 7 days. The antiproliferative effects can be determined by direct cell counts, bach ol the compounds can inhibit the growth of the cells by 30-80% at concentrations of 0.1

ng/niL.

Example 3

Effect of modified heptanoic acids of the present invention on human colorectal cancer rumor growth in male NCr-nu athymic nude mice may be observed as described in (Klyosov, Platt and Zomer, Preclimca, v. 4, 175-186, 2003)(hereiπ incorporated by reference). Briefly, thirty- to -forty .milligram fragments from an />? vivo passage of COIX) 205 human colon tumor are implanted s.c. in mice near the right axillary area using a !2-gauge trocar needle. Tumors are allowed to reach 80-200 mm' in size/volume before the start of treatment. A sufficient number of mice were implanted, so that tumors in a volume range as narrow as possible may be selected for the trial on the day of treatment initiation (day 14 after tumor implantation). Animals selected with tumors in

the proper size range are divided into the various treatment groups of 10 mice in each. The median tumor volumes in each treatment group range from 90 to 120 rnπv\ Untreated control tumors may grow well in all mice, with the median number of days for quadrupling the tumor volume equal to 13 days. There is no tumor regression after 50 days of the study, and there may be practically no tumor reduction. Median tumor volume may increase from 1 10 rrmr at treatment initiation (with saline only) to 1900 mm* after 7 weeks. Mean survival time may be equal to 14 days. At an Lv. dosage of 6 mg/kg/dose of

modified iieptanoie acids of the present invention injected daily four days in. a row, the median numbers of days far quadrupling the tumor volume is at least 46 days, with mean survival time potentially shifting from 14 days to 46 days.

Example 4

Suppression of estrogen deficiency bone loss in ovarieetomized rat model in vivo (a model of posϊ-menopausai osteoporosis) is studied in 5-month~oki OVX female rats.

49 treated orally with doses of modified heplanoie acids of the present invention ( 10 rag/kg/day daily for 4 weeks). The treatment begins 1 day after surgery. Saline-treated OVX rats (6 rats per group) serve as controls. Effects of the compounds on bone loss are measured as described in U.S. Pat. No. 6,756,401 by measuring serum osteocalcin, a biochemical marker of bone turnover, by radioimmunoassay, 80% increase of serum osteocalcin concentration is observed 4 weeks after surgery in the control, while serum osteocalcin increase is oracticallv uπnoticeabie in the treated animals.

Example 5

Suppression of cholesterol levels in New-Zealand while rabbits is studied by oral adminismition of modified heptanoic acids of the present invention into their diet and then .measuring cholesterol level in the animal plasma. Female rabbits, age 3-4 months, six per group, are fed a control diet (rabbit chow) containing 0.2% cholesterol, or an experimental diet, containing 0.2% cholesterol (200 mg/100 g) and 0.002% of a glycosylated compound (2 mg/100 g). Alter 7 weeks, plasma is collected from the rabbits and cholesterol level Isdeteraiined.

In yet another embodiment, the therapeutical Sy effective amount of modified heptanoic acids of the present invention may be in the form of a liquid, a powder and/or an aerosol. These forms of administration of modified heptanoic acids of the present invention may utilize but are not limited to the dosage regimen and route of administration already established and approved for the therapeutic agent, the difference

50 being the inclusion of a carbohydrate group into She modified heptanok acids of the present Invention. For example, a single bolus can be administered, several divided doses can be administered over a period of time, or a dose can bε proportionally reduced and administered over a time period by infusion, or can be increased, as indicated by the

exigencies of Che therapeutic situation.

In yet another embodiment, the modified heptanoie acids of the present kiveotiosi and another therapeutic agent is administered sequentially as distinct formulations.

In another example, the formulation of the mixture may be derived from fee standard formulation of the therapeutic agent to which modified heptanoic acids of the present

invention are added in a compatible solvent or as a powder,

In yet another embodiment, pharmaceutically acceptable carriers are added. For example in oral formulations, suitable pharmaceutically acceptable carriers include, but

are not limited to, hydroxy propyl cellulose, colloidal silicon dioxide, magnesium carbonate, methaerylic acid copolymer, starch, talc, sugar sphere, sucrose, polyethylene glycol polysorbate 80, and titanium dioxide: croscamieilose sodium, edible inks, gelatin, lactose rnαπohydrate, magnesium stearaie, povidone, sodium Iauryi sulfate, camuba wax, crøspovideme. hydroxy-propyl methyleeliuiose. lactose, mierocrystelliπe cellulose, and

other ingredients may be used.

One of ordinary skill in the an can determine and prescribe the effective dose of the therapeutic composition required based on clinical protocols. Io general, a suitable daily close of a composition of the invention will be that amount of the composition, which is the lowest dose effective to produce a therapeutic effect,

in another embodiment, the use of the modified heplanoic acids of the present invention containing formulation can have an immediate effect of increasing ihe responses of patients to treatment utilizing the modified heptanoic acids of the present invention

treatment, for example, by decreasing in the dosage of the agent required for effective therapy, in the presence of the formulation. It can have an immediate beneficial effect for the patient by decreasing side effects of the drugs, and thereby improving a patient's

quality of Hfe,

The use of the modified heptanoic acids of the present invention administered in & suitable formulation can be applied to a wide range of agents and is not restricted to ants- hyperlipidernie, anti-hypercholcstcroiaemic. anti-tumor, anti-cancer or ami -metastatic agents. Therapeutic areas include anti-depressants, anti-mtiammatury agents, gastroenterology drugs (for treating ulcers and associated disorders), anti-psychotic drugs, agents, etc. As many therapeutic agents must he administered as & chronic medicine, i.e., oa a long-term basis, potential reduction in dosage and improvement in

quality of life become significant factors in availability,, cost of therapeutic agents, and

patient compliance. In yel another embodiment, the modified heptanoic acids of the present invention can be administered in any of the methods known in the art such as in a liquid formulation, tablet suppository, gel, cream, transdermal or iopicai patch or aerosol. The formulation may be administered to a subject by any of the routes known in the art including by oral,

mucosal inhalation, intravenous, intramuscular, or by parenteral administration as

defined above.

Claims

What is claimed is:

1. A compound having the formula (I)_* wherein R is selected from the group consisting of heterocyclic, aromatic, and cyclic

compounds; wherein the dashed line in Formula I is selected from the group consisting of single

and/or double carbon-carbon bonds;

and wherein at least one of R/ or R^" is a carbohydrate.

HOv

N\\QR"

_;0R'

..^

(Formula 1)

2. The compound according to claim 1, wherein at least one of R" and R"' is a

monosaccharide.

3. The compound according to claim. L wherein at least one of R^" and R" is a

galactose.

4. The compound according to claim L wherein R is selected from the aromatic group consisting of condensed aromatic, substituted aromatic, hcxahydronaphthaleπe, and aromatic derivative compounds.

54

5. The compound according to claim 1, wherein R is selected from the heterocyclic group consisting of substituted heterocyclic, heterocyclic derivative, aromatic heterocyclic, nitroaromatic heterocyclic, indole.

pyrrole, pyridine, and pyrimldine compounds.

6. The compound according to claim I , wherein R is selected from the cyclic group consisting of cyclic, substituted cyclic, and cyclic derivative

compounds.

7. ^'The compound according to claim 4, wherein R is further comprised of a

side chain selected from ihe group consisting of hutanoic acid, 2- rnethylbuiarioie acid, or 2-dimeihyIbuianoic acid.

S, The compound according to claim 1. wherein R is the following compound,

and wherein the dashed line represents a chemical bond between R and the

compound according to Claim 1 :

G¹

(Formula 4)

9, The compound according to claim 8, wherein X is a monosaccharide.

10, The compound according to claim S₅ wherein X is a galactose.

11 , A compound having the formula 1, wherein the compound is a lactone.