Summary of the invention
In order to overcome the deficiencies in the prior art, the object of the present invention is to provide a kind of new bit esterified camptothecin derivative and isomer thereof of 20-;
Another object of the present invention is to provide a kind of new 20-the preparation method of bit esterified camptothecin derivative;
One aspect of the present invention relates to pharmaceutical composition, and it comprises carrier commonly used in the compound of the general formula (I) as active ingredient and isomer and the pharmacy field.
Another object of the present invention is to provide the application of bit esterified camptothecin derivative of a kind of new 20-and composition thereof as antitumor drug.
First aspect of the present invention relates to the compound shown in general formula (I).
In this general formula
R
1Be selected from: hydrogen, halogen, low alkyl group, lower alkoxy, junior alkyl halides, hydroxyl, RCOO, cyano group, aldehyde radical, nitro, amino, carboxyl, lower alkoxycarbonyl, rudimentary alcoxyl carbonyl amino, lower alkyl aminomethyl, (4-methyl isophthalic acid-piperazinyl) methyl, the lower trialkyl silylation, alkane carbonyl oxygen methyl, benzene methoxyl methyl, benzene first carbonyl oxygen methyl, rudimentary alkoxyl-methyl, CH
2NR
5R
6(R
5And R
6Be respectively hydrogen, the 1-6 carbon atom alkyl, the aryl substituted alkyl, hydroxyl replaces low alkyl group, the amino low alkyl group that replaces, single or two substituted-amino low alkyl groups, ring is amino);
R
2Be selected from: hydrogen, halogen, low alkyl group, lower alkoxy, hydroxyl, RCOO, cyano group, aldehyde radical, nitro, amino, RCONH, junior alkyl halides, carboxyl, lower alkoxycarbonyl, rudimentary alcoxyl carbonyl amino, CH
2NR
7R
8(R
7And R
8Be respectively hydrogen, the 1-6 carbon atom alkyl, the aryl substituted alkyl, hydroxyl replaces low alkyl group, the amino low alkyl group that replaces, single or two substituted-amino low alkyl groups, ring is amino), CH
2R
9(R
9Be lower alkoxy, cyano group), NR
10R
11(R
10, R
11Be respectively low alkyl group, aryl, dialkylamino alkyl, amino low alkyl group, hydroxyl low-grade alkyl);
R
3Be selected from: hydrogen, halogen, low alkyl group, lower alkoxy, amino lower alkoxy, hydroxyl, the RCOO of replacing, cyano group, aldehyde radical, nitro, amino, junior alkyl halides, carboxyl, lower alkoxycarbonyl, rudimentary alcoxyl carbo methoxy group, rudimentary alcoxyl carbonyl amino, the amino low alkyl group that replaces, hydroxyl replaces low alkyl group, [4-(piperidino)-piperidino] carbonyl oxygen base;
R
4Be selected from: hydrogen, halogen, low alkyl group, lower alkoxy, RCOO, cyano group, aldehyde radical, nitro, amino, junior alkyl halides, carboxyl, lower alkoxycarbonyl, rudimentary alcoxyl carbonyl amino;
Or R
1And R
2Cheng Huan, or R
2And R
3Cheng Huan, R
3And R
4Cheng Huan;
It is characterized in that:
R is: the 1-pyrimidyl (I) of a.1-pyrimidyl or replacement;
R
5Be: hydrogen, halogen, low alkyl group.
B. benzene sulfydryl or substituted benzene sulfydryl (II);
R
6Be: hydrogen, halogen, nitro, amino, low alkyl group, lower alkoxy, cyano group, the list of aldehyde radical replaces or intersects polysubstituted.
C. pyridine sulfydryl or substituted pyridines sulfydryl (III);
R
7Be: hydrogen, halogen, nitro, amino, low alkyl group, lower alkoxy, cyano group, the list of aldehyde radical replaces or intersects polysubstituted, nitrogen-atoms again aromatic ring 2,3,4,5,6.
D. pyrimidine sulfydryl or substituted pyrimidines sulfydryl (IV);
R
8Be: hydrogen, halogen, nitro, amino, low alkyl group, lower alkoxy, cyano group, the list of aldehyde radical replaces or intersects polysubstituted, and the sulfydryl position can be at 2,4 or 5.
E. aryl or substituted aryl (V);
R
9Be: hydrogen, halogen, nitro, amino, low alkyl group, lower alkoxy, cyano group, aldehyde radical, the list of aryl replaces or intersects polysubstituted.
F. array structure (VI) under being:
R
10Be: hydrogen, halogen, nitro, amino, low alkyl group, lower alkoxy, cyano group, the list of aldehyde radical replaces or intersects polysubstituted.
According to the present invention, the compound of preferred general formula (I) is including but not limited to the compound shown in general formula (Ia):
Wherein, R as defined above.
According to the present invention, the compound of preferred general formula (I) is including but not limited to the compound shown in general formula (Ib):
R
11Be: hydrogen, low alkyl group, lower alkylcarbonyl.
Wherein, R as defined above.
According to the present invention, the compound of preferred general formula (I) is including but not limited to the compound shown in general formula (Ic):
Wherein, R as defined above.
In the present invention, term " halogen " is meant fluorine, chlorine, bromine, iodine.Term " low alkyl group " " lower alkyl " is the alkyl of the straight or branched of 1-6 carbon atom.
According to the present invention, the form that The compounds of this invention can isomer exists, and described usually " The compounds of this invention " comprises the isomer of this compound.
Can there be the cis-trans-isomer of two keys in The compounds of this invention, and asymmetric center has S configuration or R configuration, the present invention includes all possible steric isomer and two or more mixture of isomers.If there is suitable/trans isomer, the present invention relates to the mixture of cis form and trans forms and these forms, single if desired foreign body object can separate according to conventional methods or prepare by three-dimensional selection is synthetic.
According to embodiment of the present invention, described The compounds of this invention also comprises hydrate, ester or the prodrug of acceptable salt, salt on its pharmacodynamics.
Another aspect of the present invention relates to the preparation method of The compounds of this invention.
The compound of invention is to prepare by reacting camptothecine or its analogue and corresponding carboxylic acid or carboxylic acid halides.Camptothecin analogues is that in camptothecine 7,9,10,11 have various substituent compounds, and it is synthetic that their preparation can be consulted literature method.Provide the title of part document below.
Reference:
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Preferred acid be selected from 2-(ethyl thioglycollic acid of pyridine-4-), 2-(thymus pyrimidine-1-) acetate, 2-[(4-methylpyrimidine)-2-] ethyl thioglycollic acid, 3,4,5-trimethoxy toluylic acid, 3-(4-benzoyl bromide) propionic acid, 4-chlorobenzene ethyl thioglycollic acid.The mol ratio of acid and camptothecine is 1-5: 1; Preferred 1.5-3.5: 1; More preferably 2-3: 1.
Condensation reaction takes place under the condition that suitable condensing agent, catalyzer exist.Preferred condensation reagent is to comprise 1,3 dicyclohexylcarbodiimide (DCC), two pyridine carbonic ethers (2-DPC), 1-(3-dimethylamine propyl)-3-ethyl carbimide hydrochloride (EDCI), 1,3-di-isopropyl carbimide (DIPC), 4-pyrrolidyl pyridine.1-[(3-dimethylamino most preferably) propyl group]-3-ethyl carbimide hydrochloride (EDCI), the 1-[(3-dimethylamino) propyl group]-mol ratio of 3-ethyl carbimide hydrochloride and camptothecine is 2-6: 1; Preferred 2.5-5.5: 1; More preferably 3.5-4.5: 1.
Preferred catalyzer is a tertiary amine.Preferred tertiary amine is selected from 4-Dimethylamino pyridine (DMAP), 4-pyrrolidyl pyridine.4-Dimethylamino pyridine (DMAP) most preferably, the mol ratio of (0.71,0.95) 4-Dimethylamino pyridine and camptothecine is 0.5-1.5: 1; Preferred 0.75-0.95: 1.
Be reflected under the The suitable solvent and carry out, preferred solvent is anhydrous non-protonic solvent, more preferably haloalkane, DMF, tetrahydrofuran (THF) (THF), N, dinethylformamide (DMF) etc.Methylene dichloride most preferably.
Be reflected at-carry out under the 20-60 ℃, preferred temperature of reaction is between 0 ℃-60 ℃, and preferred temperature is 20 ℃-30 ℃, and most preferred temperature is a room temperature.
Reaction times is 2-48 hour, preferably 6-36 hour, and most preferably 10-24 hour.
With this neighborhood technician known method washing, dry, concentrated, separate with silica gel column chromatography, eluent is chloroform and methanol mixture.
Further aspect of the present invention also relates to and containing as the The compounds of this invention of active ingredient and the pharmaceutical composition of conventional medicine vehicle or assistant agent.Usually pharmaceutical composition of the present invention contains the The compounds of this invention of 0.1-95 weight %.
The pharmaceutical composition of The compounds of this invention can be according to method preparation well known in the art.When being used for this purpose, if desired, The compounds of this invention The compounds of this invention and one or more solids or liquid medicine vehicle and/or assistant agent can be combined, make and can be used as suitable administration form or the dosage form that people's medicine or veterinary drug use.
The compounds of this invention or contain its pharmaceutical composition can the unit dosage form administration, route of administration can be enteron aisle or non-enteron aisle, as oral, muscle, subcutaneous, nasal cavity, oral mucosa, skin, peritonaeum or rectum etc.
The compounds of this invention or the route of administration that contains its pharmaceutical composition can be drug administration by injection.Injection comprises intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection and acupoint injection therapy etc.
Form of administration can be liquid dosage form, solid dosage.As liquid dosage form can be true solution class, colloidal type, particulate formulations, emulsion dosage form, mixed suspension form.Other formulations are tablet, capsule, dripping pill, aerosol, pill, pulvis, solution, suspensoid, emulsion, granule, suppository, lyophilized injectable powder etc. for example.
The compounds of this invention can be made ordinary preparation, also can be sustained release preparation, controlled release preparation, targeting preparation and various particulate delivery system.
For example, can be extensive use of various carrier well known in the art for the unit form of administration is made tablet.Example about carrier is, for example thinner and absorption agent are as starch, dextrin, calcium sulfate, lactose, N.F,USP MANNITOL, sucrose, sodium-chlor, glucose, urea, lime carbonate, white bole, Microcrystalline Cellulose, pure aluminium silicate etc.; Wetting agent and tackiness agent are as water, glycerine, polyoxyethylene glycol, ethanol, propyl alcohol, starch slurry, dextrin, syrup, honey, glucose solution, mucialga of arabic gummy, gelatine size, Xylo-Mucine, lac, methylcellulose gum, potassiumphosphate, polyvinylpyrrolidone etc.; Disintegrating agent, for example dry starch, alginates, agar powder, laminaran, sodium bicarbonate and Citric Acid, lime carbonate, polyoxyethylene sorbitol fatty acid ester, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.; Disintegration inhibitor, for example sucrose, Tristearoylglycerol, theobroma oil, hydrogenation wet goods; Absorption enhancer, for example quaternary ammonium salt, sodium lauryl sulphate etc.; Lubricant, for example talcum powder, silicon-dioxide, W-Gum, stearate, boric acid, whiteruss, polyoxyethylene glycol etc.Tablet further can also be made coating tablet, for example sugar coated tablet, thin membrane coated tablet, ECT, or double-layer tablets and multilayer tablet.
For example, can be extensive use of various carrier well known in the art for pill is made in the administration unit.Example about carrier is, for example thinner and absorption agent are as glucose, lactose, starch, theobroma oil, hydrogenated vegetable oil, polyvinylpyrrolidone, Gelucire, kaolin, talcum powder etc.; Tackiness agent is as gum arabic, tragacanth gum, gelatin, ethanol, honey, liquid sugar, rice paste or batter etc.; Disintegrating agent is as agar powder, dry starch, alginates, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.
For example for capsule is made in the administration unit, the effective constituent The compounds of this invention is mixed with above-mentioned various carriers, and the mixture that will obtain thus places hard gelatine capsule or soft capsule.Also the effective constituent The compounds of this invention can be made microcapsule, be suspended in and form suspensoid in the aqueous medium, in the hard capsule of also can packing into or make injection and use.
For example, The compounds of this invention is made injection preparation, as solution, suspensoid solution, emulsion, lyophilized injectable powder, this preparation can be moisture or non-water, can contain acceptable carrier, thinner, tackiness agent, lubricant, sanitas, tensio-active agent or dispersion agent on a kind of and/or multiple pharmacodynamics.Can be selected from water, ethanol, polyoxyethylene glycol, 1 as thinner, the isooctadecanol of ammediol, ethoxylation, the isooctadecanol of polyoxyization, Polyoxyethylene Sorbitol Fatty Acid Esters etc.In addition, ooze injection liquid, can in injection preparation, add proper amount of sodium chloride, glucose or glycerine, in addition, can also add conventional solubility promoter, buffer reagent, pH regulator agent etc. in order to prepare etc.These auxiliary materials are that this area is commonly used
In addition, as needs, also can in pharmaceutical preparation, add tinting material, sanitas, spices, correctives, sweeting agent or other material.
Another aspect of the present invention also relates to the purposes of The compounds of this invention and pharmaceutical composition.The dosage of The compounds of this invention and pharmaceutical composition depends on many factors, for example to prevent or treat the character and the severity of disease, the sex of patient or animal, age, body weight, personality and individual reaction, route of administration, administration number of times etc., therefore therapeutic dose of the present invention can have large-scale variation.In general, the using dosage of Chinese materia medica composition of the present invention is well known to a person skilled in the art.Can be according to the actual drug quantity that is contained in the preparation last in the The compounds of this invention composition, in addition suitable adjustment to reach the requirement of its treatment significant quantity, is finished prevention of the present invention or therapeutic purpose.The suitable dose scope of the every day of The compounds of this invention is from 1-1000mg/m
2, 10-100mg/m especially
2Above-mentioned dosage can the single dose form or be divided into several, two, three or four these dosage regimens that are subject to administration doctor's clinical experience and comprise utilization chemotherapy, radiotherapy means of dosage form administration for example.
The invention compound by suitable medication such as oral, parenterai administration such as IP, IV treats animal tumor.Tumour comprises leukemia and solid tumor such as colon, ovary, mammary gland, prostate gland, lung, kidney and melanoma.The dosage range of administration depends on route of administration, age, body weight, body surface area and patient's treatment situation.Concerning the people, body surface area is determined by height and body weight.The suitable dose scope of the compound of invention is from 1-1000mg/m
2, 10-100mg/m especially
2
Abbreviation
EDCI (1-3-dimethylamine propyl)-3-ethyl carbimide hydrochloride;
DCC:1,3-dicyclohexyl carbimide;
DIPC:1,3-di-isopropyl carbimide;
The DMAP:4-dimethylamino pyridine;
THF: tetrahydrofuran (THF);
DMF:N, dinethylformamide.
Embodiment
The following examples are used for further specifying the present invention, but this and do not mean that any limitation of the invention.
Embodiment 1.
Camptothecine-20-O-2-(ethyl thioglycollic acid ester of pyridine-4-)
In the dry round-bottomed flask of the 25mL that has induction stirring, add 30mg (0.0861mmol) camptothecine, 30mg (0.172mmol) 2-(pyridine-4-) ethyl thioglycollic acid, 66mg (0.344mmol) 1-[(3-dimethylamino) propyl group]-3-ethyl carbimide hydrochloride (EDCI), 10mg (0.0819mmol) 4-Dimethylamino pyridine (DMAP) and 5mL anhydrous methylene chloride, reaction mixture was stirring at room 10 hours, reaction solution with the dilution of 50mL chloroform, is used 50mL water, the saturated NaHCO of 50mL then successively
3Solution and the water washing of 50mL saturated common salt add anhydrous MgSO
4Drying removes by filter MgSO
4After, the pressure reducing and steaming solvent, residue with silica gel column chromatography separate (eluent: chloroform-methanol 100: 1), obtain the 26mg white solid, yield: 60%, mp:207-210 ℃,
1H NMR (400MHZ, CDCl
3): δ 8.41 (s, 1H, Ar-H), 8.24 (m, 3H, Ar-H), 7.97 (d, J=8.0Hz, 1H, Ar-H), 7.88 (t, J=7.2Hz, 1H, Ar-H), 7.70 (t, J=7.2Hz, 1H, Ar-H), 7.35 (d, J=5.6Hz, 2H, Ar-H), 7.17 (s, 1H, Ar-H), 5.70 (d, J=17.2Hz, 1H, H17), 5.41 (d, J=17.2Hz, 1H, H17), 5.27 (s, 2H, H5), 3.97 (s, 2H, SCH
2), 2.28 (dm, 2H, 19-CH
2), 0.98 (t, J=7.6Hz, 3H, 18-CH
3).
Embodiment 2.
Camptothecine-20-O-2-(acetic ester of thymus pyrimidine-1-)
In the dry round-bottomed flask of the 25mL that has induction stirring, add 40mg (0.106mmol) camptothecine, 60mg (0.326mmol) 2-(thymus pyrimidine-1-) acetate, 80mg (0.424mmol) 1-[(3-dimethylamino) propyl group]-3-ethyl carbimide hydrochloride (EDCI), 10mg (0.0819mmol) 4-Dimethylamino pyridine (DMAP) and 5mL anhydrous methylene chloride, reaction mixture was stirring at room 24 hours, reaction solution with the dilution of 50mL chloroform, is used 50mL water, the saturated NaHCO of 50mL then successively
3Solution and the water washing of 50mL saturated common salt add anhydrous MgSO
4Drying removes by filter MgSO
4After, the pressure reducing and steaming solvent, residue with silica gel column chromatography separate (eluent: chloroform-methanol 100: 1), obtain the 14mg yellow solid, yield: 24%, mp:206-208 ℃,
1H NMR (400MH
Z, CDCl
3): δ 8.50 (s, 1H, Ar-H), 8.44 (d, J=8.0Hz, 1H, Ar-H), 8.00 (d, J=8.0Hz, 1H, Ar-H), 7.91 (t, J=7.6Hz, 1H, Ar-H), 7.74 (t, J=7.6Hz, 1H, Ar-H), 7.58 (s, 1H, Ar-H), 7.14 (s, 1H, Ar-H), 5.67 (d, J=17.6Hz, 1H, H17), 5.42 (d, J=17.6Hz, 1H, H17), 5.32 (s, 2H, H5), 4.89 (d, J=17.6Hz, 1H, NCH
2COO), 4.59 (d, J=17.6Hz, 1H, NCH
2COO), 2.28 (dm, 2H, 19-CH
2), 1.01 (t, J=7.6Hz, 3H, 18-CH
3).
Embodiment 3.
Camptothecine-20-O-2-[(4-methylpyrimidine)-and 2-] the ethyl thioglycollic acid ester
In the dry round-bottomed flask of the 25mL that has induction stirring, add 40mg (0.115mmol) camptothecine, 52mg (0.282mmol) 2-[(4-methylpyrimidine)-2-] ethyl thioglycollic acid, 90mg (0.469mmol) 1-[(3-dimethylamino) propyl group]-3-ethyl carbimide hydrochloride (EDCI), 10mg (0.0819mmol) 4-Dimethylamino pyridine (DMAP) and 5mL anhydrous methylene chloride, reaction mixture was stirring at room 12 hours, reaction solution with the dilution of 50mL chloroform, is used 50mL water, the saturated NaHCO of 50mL then successively
3Solution and the water washing of 50mL saturated common salt add anhydrous MgSO
4Drying removes by filter MgSO
4After, the pressure reducing and steaming solvent, residue with silica gel column chromatography separate (eluent: chloroform-methanol 100: 1), obtain the 50mg yellow solid, yield: 85%, mp:231-233 ℃,
1H NMR (400MH
Z, CDCl
3): δ 8.40 (s, 1H, Ar-H), 8.21 (d, J=8.0Hz, 1H, Ar-H), 7.97 (d, J=8.0Hz, 1H, Ar-H), 7.86 (t, J=7.6Hz, 1H, Ar-H), 7.69 (t, J=7.6Hz, 1H, Ar-H), 7.25 (s, 1H, Ar-H), 6.60 (s, 1H, Ar-H), 5.62 (d, J=17.2Hz, 1H, H17), 5.43 (d, J=17.2Hz, 1H, H17), 5.27 (s, 2H, H5), 4.12 (d, J=16.8Hz, 1H, SCH
2), 4.08 (d, J=16.8Hz, 1H, SCH
2), 2.34 (m, 3H NCCH
3), 2.22 (dm, 2H, 19-CH
2), 0.95 (t, J=7.6Hz, 3H, 18-CH
3).
Embodiment 4.
Camptothecine-20-O-3,4,5-trimethoxy phenylacetate
In the dry round-bottomed flask of the 25mL that has induction stirring, add 40mg (0.115mmol) camptothecine, 65mg (0.282mmol) 3,4,5-trimethoxy toluylic acid, 90mg (0.469mmol) 1-[(3-dimethylamino) propyl group]-3-ethyl carbimide hydrochloride (EDCI), 10mg (0.0819mmol) 4-Dimethylamino pyridine (DMAP) and 5mL anhydrous methylene chloride, reaction mixture was stirring at room 12 hours, reaction solution with the dilution of 50mL chloroform, is used 50mL water, the saturated NaHCO of 50mL then successively
3Solution and the water washing of 50mL saturated common salt add anhydrous MgSO
4Drying removes by filter MgSO
4After, the pressure reducing and steaming solvent, residue with silica gel column chromatography separate (eluent: chloroform-methanol 100: 1), obtain the 52mg yellow solid, yield: 82%, mp:216-219 ℃,
1H NMR (400MHZ, CDCl
3): δ 8.39 (s, 1H, Ar-H), 8.25 (d, J=8.0Hz, 1H, Ar-H), 7.93 (d, J=8.0Hz, 1H, Ar-H), 7.85 (t, J=7.2Hz, 1H, Ar-H), 7.68 (t, J=7.2Hz, 1H, Ar-H), 7.14 (s, 1H, Ar-H), 6.52 (s, 2H, Ar-H), 5.65 (d, J=17.2Hz, 1H, H17), 5.42 (d, J=17.2Hz, 1H, H17), 5.27 (s, 2H, H5), 3.77 (m, 9H, OCH
3), 2.29 (dm, 2H, 19-CH
2), 0.98 (t, J=7.6Hz, 3H, 18-CH
3).
Embodiment 5.
Camptothecine-20-O-3-(4-benzoyl bromide) propionic ester
In the dry round-bottomed flask of the 25mL that has induction stirring, add 30mg (0.0861mmol) camptothecine, 44mg (0.190mmol) 3-(4-benzoyl bromide) propionic acid, 60mg (0.313mmol) 1-[(3-dimethylamino) propyl group]-3-ethyl carbimide hydrochloride (EDCI), 10mg (0.0819mmol) 4-Dimethylamino pyridine (DMAP) and 5mL anhydrous methylene chloride; reaction mixture was stirring at room 12 hours; reaction solution with the dilution of 50mL chloroform, is used 50mL water, the saturated NaHCO of 50mL then successively
3Solution and the water washing of 50mL saturated common salt add anhydrous MgSO
4Drying removes by filter MgSO
4After, the pressure reducing and steaming solvent, residue with silica gel column chromatography separate (eluent: chloroform-methanol 100: 1), obtain the 44mg yellow solid, yield: 87%, mp:215-218 ℃, IR (KBr) ν 3485,2939,2821,1743,1662,1616,1500,1238,1157,760,723,592cm
-1 1H NMR (600MHZ, CD
3Cl
3): δ 8.40 (s, 1H, Ar-H), 8.28 (d, J=8.4Hz, 1H, Ar-H), 7.95 (d, J=8.4Hz, 1H, Ar-H), 7.81 (t, J=7.2Hz, 1H, Ar-H), 7.79 (d, J=7.8Hz, 2H, Ar-H), 7.69 (t, J=7.2Hz, 1H, Ar-H), 7.48 (d, J=7.8Hz, 2H, Ar-H), 7.41 (s, 1H, Ar-H), 5.67 (d, J=16.8Hz, 1H, H17), 5.41 (d, J=16.8Hz, 1H, H17), 5.29 (d, J=10.8Hz, 2H, H5), 3.25 (m, 2H, ArCOCH
2), 3.03 (m, 2H, ArCOCCH
2), 2.18 (dm, 2H, 19-CH
2), 1.01 (t, J=7.2Hz, 3H, 18-CH
3).
Embodiment 6
Camptothecine-20-O-4-chlorobenzene ethyl thioglycollic acid ester
In the dry round-bottomed flask of the 25mL that has induction stirring, add 30mg (0.0861mmol) camptothecine, 30mg (0.148mmol) 4-chlorobenzene ethyl thioglycollic acid, 60mg (0.313mmol) 1-[(3-dimethylamino) propyl group]-3-ethyl carbimide hydrochloride (EDCI), 10mg (0.0819mmol) 4-Dimethylamino pyridine (DMAP) and 3mL anhydrous methylene chloride, reaction mixture was stirring at room 12 hours, reaction solution with the dilution of 50mL chloroform, is used 50mL water, the saturated NaHCO of 50mL then successively
3Solution and the water washing of 50mL saturated common salt add anhydrous MgSO
4Drying removes by filter MgSO
4After, the pressure reducing and steaming solvent, residue with silica gel column chromatography separate (eluent: chloroform-methanol 100: 1), obtain the 23mg yellow solid, yield: 50%, mp:182-184 ℃,
1H NMR (600MHZ, CD
3Cl
3): δ 8.40 (s, 1H, Ar-H), 8.24 (d, J=8.4Hz, 1H, Ar-H), 7.96 (d, J=8.4Hz, 1H, Ar-H), 7.86 (t, J=7.2Hz, 1H, Ar-H), 7.69 (t, J=7.2Hz, 1H, Ar-H), 7.39 (d, J=8.4Hz, 2H, Ar-H), 7.22 (s, 1H, Ar-H), 7.10 (d, J=8.4Hz, 2H, Ar-H), 5.65 (d, J=16.8Hz, 1H, H17), 5.41 (d, J=16.8Hz, 1H, H17), 5.26 (s, 2H, H5), 3.78 (d, J=15.0Hz, 1H, SCH
2), 3.73 (d, J=15.0Hz, 1H, SCH
2), 2.23 (dm, 2H, 19-CH
2), 0.94 (t, J=7.2Hz, 3H, 18-CH
3).
Pharmacological evaluation
Experimental example 1.MTT legal person cancer cells fragmentation test
Medicine and reagent: DMSO analytical pure; RPMI1640 is the GIBCO product; Calf serum is the animal doctor of a Beijing Military Area Command centre of prevention and cure product.
Instrument: BIORAD550 type microplate reader.
Mtt assay is measured: collect well-grown tumour cell, be mixed with 1 * 10 with the RPMI1640 substratum that contains 10% calf serum
-4/ mL cell suspension is inoculated in 96 well culture plates, and every empty 100 μ L (containing 1000 tumour cells) put 37 ℃, 5%CO
2Cultivate dosing after 24 hours in the incubator, blank and solvent control are established in experiment, and given the test agent is established 4 concentration, and 3 parallel holes of every concentration are put 37 ℃, 5%CO
2Cultivated 4 days in the incubator.Discard nutrient solution, every hole adds MTT solution (0.4mg/mL, RPMI1640 preparation) 100 μ L, hatches 4 hours for 37 ℃.Abandoning supernatant, every hole add MTT solution 150 μ L, and dissolved particles behind the gentle agitation, is detecting wavelength 540nm with 550 type microplate reader, and reference wavelength 450nm measures the OD value down.
The result calculates: can obtain dose response curve with the different concns of medicine and the inhibiting rate mapping of pair cell, therefrom obtain half-inhibition concentration (IC
50).
Derivative of the present invention the results are shown in Table 1. to the anti-tumor activity of human tumor cell line
The anti tumor activity in vitro of table 1 camptothecin derivative
Compound | IC
50/μmol·L-1
|
KB | KB/VCR | A549 | HCT-8 | Bel7402 | A2780 |
Topotecan CPT 1 embodiment 12 embodiment 23 embodiment 34 embodiment 45 embodiment 56 embodiment 6 | 0.007 0.003 0.006 0.005 0.033 0.014 0.042 0.049 | 0.100 0.031 0.511 0.080 0.924 0.500 0.357 0.204 | 0.078 0.009 0.076 0.009 0.245 0.086 0.306 0.313 | 0.022 0.003 0.008 0.005 0.551 0.010 0.087 0.055 | 0.078 0.007 0.050 0.008 0.100 0.153 0.090 0.087 | 0.032 0.006 0.044 0.007 0.0815 0.063 0.093 0.072 |
KB: human oral cancer cells; KB/VCR: people's resistance cancer cell of oral cavity; A549: human lung adenocarcinoma cell; A2780: Proliferation of Human Ovarian Cell; HCT-8: human colon cancer cell; Bel7402: human liver cancer cell.