CN100475835C - 23-Hydroxyl betulinic acid kind derivant and preparing process, preparation and use thereof - Google Patents

23-Hydroxyl betulinic acid kind derivant and preparing process, preparation and use thereof Download PDF

Info

Publication number
CN100475835C
CN100475835C CNB2006100402772A CN200610040277A CN100475835C CN 100475835 C CN100475835 C CN 100475835C CN B2006100402772 A CNB2006100402772 A CN B2006100402772A CN 200610040277 A CN200610040277 A CN 200610040277A CN 100475835 C CN100475835 C CN 100475835C
Authority
CN
China
Prior art keywords
acid
white birch
diacetyl
white
dihydroxyl
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CNB2006100402772A
Other languages
Chinese (zh)
Other versions
CN1861627A (en
Inventor
吴晓明
徐进宜
毕毅
周金培
张陆勇
袁貹涛
李丹
徐萍
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
China Pharmaceutical University
Original Assignee
China Pharmaceutical University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by China Pharmaceutical University filed Critical China Pharmaceutical University
Priority to CNB2006100402772A priority Critical patent/CN100475835C/en
Publication of CN1861627A publication Critical patent/CN1861627A/en
Application granted granted Critical
Publication of CN100475835C publication Critical patent/CN100475835C/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Landscapes

  • Steroid Compounds (AREA)

Abstract

A kind of modified 23-hydroxy betulic acid derivatives, their preparing process, the medicinal composition containing them and their application in preventing and treating tumor are disclosed.

Description

23-Hydroxyl betulinic acid kind derivant, its preparation method, preparation and purposes
Technical field
The present invention relates to natural drug and pharmaceutical chemistry field, be specifically related to the modified 23-hydroxyl radical white birck acid derivative of a class, the invention also discloses the preparation method of these 23-hydroxyl radical white birck acid derivatives, the pharmaceutical composition that contains these 23-hydroxyl radical white birck acid derivatives and this class 23-hydroxyl radical white birck acid derivative and be used for anti-tumor application.
Background technology
The 23-hydroxyl radical white birck acid is the compound with white birch acid structural similitude, the 23-hydroxyl radical white birck acid is a kind of pentacyclic triterpenoid that extracts from herbal medicine Root of Chinese Pulsatilla (Pulsatilla chinensis Regel), the Root of Chinese Pulsatilla beginning is stated from Shennong's Herbal, belong to Ranunculaceae Pulsatilla plant, have clearing heat and detoxicating, remove heat from the blood and relieve diarrhea, the effect of removing dampness and killing parasites.Be mainly used in treatment bacillary dysentery, amebic dysentery, gynecology colpitis etc. clinically.Modern study is found Root of Chinese Pulsatilla except antibiotic and disease-resistant protozoon effect, also has anticancer, effects such as essence, treatment endotoxemia extremely.Separate obtaining sapogenin 23-hydroxyl radical white birck acid (23-Hydroxy betulinic acid) from Root of Chinese Pulsatilla, structure and white birch acid are very close.CN 1491651A discloses the 23-hydroxyl radical white birck acid has good effect to prevention and the treatment of murine melanoma, glioma brain tumour, intestinal canal tumour, lung cancer, liver cancer and AIDS.CN1682740A discloses white birch acid and has had the glycogen phosphorylase inhibitors effect.
People carry out structure of modification to white birch acid, attempt to find the better compound of pharmacologically active, as Bioorganic; MedicinalChemistry Letters, 9 (8), a series of derivatives to white birch acid structural modification are disclosed among the 1201-1204, part-structure is as follows:
Figure C20061004027700031
This paper author has also done pharmaceutical research to this series derivates, and test-results is as follows:
Table?1.Cytotoxicity?profile?of?amino?acid?conjugates?of?betulinic?acid?against?MEL-2?and?KB?cells?lines.
Figure C20061004027700041
Note：The?assay?was?performed?twice?in?triplicates.
The?solubility?was?tested?on?the?compounds?that?showed?cytotoxicity?against?MEL-2.
From these data as can be seen the anti-tumor activity of the modified compound of part and white birch acid quite or lower slightly, the activity of part of compounds in indivedual tumour cells is higher than white birch acid, as compound 11,13,23,25.
In antitumor field, most studies person thinks that white birch acid and derivative thereof reach antineoplastic purpose by suppressing vasculogenesis, but does not still have this compounds to enter clinical experimental stage for many years at present.The clinical better antitumor drug of drug effect that needs.
Summary of the invention
The invention discloses the new 23-hydroxyl radical white birck acid derivative of a class,, obtain the 23-hydroxyl radical white birck acid derivative of general formula I by structure of modification to the 23-hydroxyl radical white birck acid:
Figure C20061004027700042
R wherein 1Represent H, Br, Cl or F;
R 2Represent OH, OR 6, OCOR 6, OSO 2R 6, OSO 2NH 2, NH 2, NHCH 2OR 6, NHR 6, N (R 6) 2, NOR 6, OCOR 6NHR 6, NHCOR 6, NHCOR 6(COOH) X, NHSO 2R 6, NHNH R 6, NHNHCH (OH) R 6, OCO R 6COOH, OCOCH (OH) CH (Ph) R 7, OCOCOOR 6, NOSO 3H, NOR 6Or NHOR 6
R 3Represent H, CH 2OH, CHO, COOH, CH 2OR 6, CH 2OCOR 6, CH 2OSO 2R 6, CH 2OCOR 6NHR 6,, CH 2NHCOR 6, CH 2NHCOR 6(COOH) X, CH 2NHSO 2R 6X, CH 2NHNHR 6, CH 2NHNHCH (OH) R 6, CH 2OCO R 6COOH, CH 2OCOCH (OH) CH (Ph) R 7, CH 2OCOCOO R 6, CH 2NOSO 3R 6, CH 2NOR 6Or CH 2NHOR 6
R 4Representative and R 3Identical group is perhaps represented COOM, COCl, CON 3, CONH 2, COOR 6, CONR 6, CON (R 6) 2, COOR 6COZ, COOR 6OX, COO (CH 2) mCH=CH (CH 2) nOR 6, COO (CH 2) mC ≡ C (CH 2) nOR 6, COOCOCH=C (R 6) 2, COOCOR 6, COOR 6OCOR 6, COOR 6OCOR 6COOH, CONHR 6NHCOR 6COOH, CONHR 6COOR 6, CONHR 6CONHR 6COOR 6, COOR 6CHO, COOCH 2CH=NOA, COOCH 2CH 2NHOA, COOCH 2CH=NR 6, COOCH 2CH 2NHNHR 6Or COOCH 2CH 2N (R 6) 2
R 5Represent CH 2=C-CH 3, CH 2=C-CH 2OH, CH 3CHCH 3Or CH 2=C-CH 2Br;
R 6The C that represents H, non-replacement or X to replace 1-C 10The alkane of straight or branched, alkene, alkynes, naphthenic hydrocarbon, phenyl, benzyl, naphthyl; And, work as R 2Be OH, R 4Be COOH or CH 2During OH, R 6Be not H;
R 7Represent NH 2Or NH R 6X;
X represents H, Cl, Br, F, I, CN, NH 2, NO 2, CF 3, OH, OCH 3, COOH, COOCH 3, C 1-C 10The alkane of straight or branched, alkene, alkynes, naphthenic hydrocarbon, phenyl, benzyl, naphthyl;
Z represents OH, OR 6, halogen, N 3Or NH 2
A represents H, CH 3, SO 2C 6H 4CH 3, OCOR 6Or R 6
M represents Na, K, Li;
m＝1-8，n＝1-8。
Preferred compound is in the said derivative:
R 1Represent H;
R 2Represent OH, OCOR 6, OCOR 6COOH ,=O, NOH, NOCOR 6, OCOCOOR 6
R 3Represent H, CH 2OH, CH 2OCOR 6, CH 2OCOR 6COOH, CH 2OCOC 6H 5, CHO, COOH, CH 2OCOCOOR 6
R 4Represent COOH, CONHR 6,
Figure C20061004027700061
COO R 6, COOR 6OH, COO (CH 2) 4OCO R 6, COOR 6OCOR 6COOH, CONHR 6NHCOR 6COOH, CONHR 6COOCH 3, CONHR 6COOH, CONHR 6CONHR 6COOR 6, CONHR 6CONHR 6COOH or CH 2OH;
R 5Represent CH 2=C-CH 3Or CH 2=C-CH 2OH;
R 6The C that represents H, non-replacement or X to replace 1-C 10The alkane of straight or branched, alkene, alkynes, naphthenic hydrocarbon, phenyl, benzyl, naphthyl; And, work as R 2Be OH, R 4Be COOH or CH 2During OH, R 6Be not H;
X represents H, Cl, Br, F, I, CN, NH 2, NO 2, CF 3, OH, OCH 3, COOH, COOCH 3, C 1-C 10The alkane of straight or branched, alkene, alkynes, naphthenic hydrocarbon, phenyl, benzyl, naphthyl;
More preferred compound is:
3,23-O-diacetyl-30-hydroxyl-28-white birch acid methyl esters
3,23-dihydroxyl-28-white birch acid acid amides glycine
3,23-dihydroxyl-28-white birch acid acid amides L-Ala
3,23-dihydroxyl-28-white birch acid acid amides L-glutamic acid
3,23-dihydroxyl-28-white birch acid acid amides Xie Ansuan
3,23-dihydroxyl-28-white birch acid acid amides leucine
3,23-dihydroxyl-28-white birch acid acid amides phenylalanine
3, the acid of 23-O-disuccinic acid monoacyl white birch
3,23-O-two phenylacetyl white birch acid
3-ketone-23-aldehyde white birch acid
3-O-phthalic acid monoacyl-23-hydroxyl-white birch acid
3-hydroxyl-23-O-benzoyl white birch acid
3,23-O-diacetyl-28-white birch acid acid amides glycine methyl ester
3,23-O-diacetyl-28-white birch acid acid amides glutamic acid dimethyl ester
3,23-O-diacetyl-28-white birch acid acid amides alanine methyl ester
3,23-O-diacetyl-28-white birch acid acid amides leucine methyl esters
3,23-O-diacetyl-28-white birch acid acid amides phenylalanine methyl ester
3,23-O-diacetyl-28-white birch acid acid amides valine methyl ester
3,23-dihydroxyl-28-white birch acid n-octyl
3, the 23-dihydroxyl-just own ester of 28-white birch acid
3, the 23-dihydroxyl-positive heptyl ester of 28-white birch acid
3,23-dihydroxyl-28-white birch acid cyclohexyl
3,23-dihydroxyl-28-white birch acid propylene ester
3,23-dihydroxyl-28-white birch acid propynyl ester
3,23-dihydroxyl-28-white birch acid butyleneglycol monoesters
3,23-dihydroxyl-28-white birch acid ethylene glycol ester
3,23-dihydroxyl-28-white birch acid hexylene glycol monoesters
3,23-dihydroxyl-28-white birch acid butynediol monoesters
3, the 23-O-diacetyl-positive butyramide of 28-white birch acid
3,23-O-diacetyl-28-white birch acid Isopropamide
3,23-O-diacetyl-28-white birch acid Cyclohexamide
3,23-O-diacetyl-28-white birch acid DB
3,23-O-diacetyl-28-white birch acid phenylacetamide
3,23-O-diacetyl-28-white birch acid diethylamide
3,23-O-diacetyl-28-white birch acid (3, the 4-(methylenedioxy)) phenylacetamide
3,23-O-diacetyl-28-white birch acid (Alpha-Methyl) benzyl acid amides
3,23-O-diacetyl-28-white birch acid methyl esters
3,23-O-diacetyl-28-white birch acid isobutyl ester
3,23-O-diacetyl-28-white birch isopropyl propionate
3,23-O-diacetyl-28-white birch acid propylene ester
3,23-O-diacetyl-28-white birch acid propynyl ester
3,23-O-diacetyl-28-white birch acid ethylene glycol ester
3,23-O-diacetyl-28-white birch acid butyleneglycol monoesters
3,23-O-diacetyl-28-white birch acid butynediol monoesters
3,23-O-diacetyl-28-white birch acid hexylene glycol monoesters
3,23-O-diacetyl-28-white birch acid butyleneglycol Succinic Acid dibasic acid esters
3,23-O-diacetyl-28-white birch acid butyleneglycol pentanedioic acid dibasic acid esters
3,23-O-diacetyl-28-white birch acid hexylene glycol pentanedioic acid dibasic acid esters
3,23-O-diacetyl-28-white birch acid ethylene glycol Succinic Acid dibasic acid esters
3,23-O-diacetyl-28-white birch acid ethylene glycol pentanedioic acid dibasic acid esters
3,23-O-diacetyl-28-white birch acid ethylene glycol bis phthalate
3,23-O-diacetyl-28-white birch acid hexylene glycol Succinic Acid dibasic acid esters
3,23-O-diacetyl-28-white birch acid ethylene glycol maleic acid dibasic acid esters
3,23-dihydroxyl-28-white birch acid acid amides glycine acid amides glycine methyl ester
23-hydroxyl radical white birck acid derivative of the present invention can prepare (R wherein with following method 2, R 3, R 4, R 6Definition before described):
Work as R 2Be OCOR 6Or OCOR 6COOH ,=O; R 3Be CH 2OCOR 6, CH 2OCOR 6COOH, CH 2OCOC 6H 5, when CHO or COOH, preparation process comprises:
A, with big excessive acid anhydrides or acyl chlorides or oxidizer treatment 23-hydroxyl radical white birck acid 4-48 hour, temperature is a room temperature to 120 ℃;
B, in the solution of step a, add entry;
C, use organic solvent extraction, obtain organic layer,
Wherein, oxygenant is PCC oxygenant (pyridinium chlorochromate) or Jone ' s oxygenant (H 2CrO 4/ H 2SO 4/ acetone), organic solvent is methylene dichloride, chloroform or ethyl acetate.
Work as R 2Be OCOR 6R 3Be CH 2OCOR 6R 4Be COOR 6OH, COO (CH 2) 4OCOR 6Or COOR 6OCOR 6During COOH, preparation process comprises:
A, with 3,23-O-diacyl-17-acyl chlorides betulinic acid derivative is dissolved in the glycol, reacts 1-48 hour, obtains 3,23-O-diacyl-28-white birch acid diol monoester derivative;
B, a product is dissolved in the halogenated organic solvent; add acid anhydrides or dicarboxylic anhydride and organic bases; reacted 1-48 hour; obtain 3; 23-O-diacyl-28-white birch acid glycol list acid diester or 3; 23-O-diacyl-28-white birch acid glycol two acid diesters, wherein halogenated organic solvent is methylene dichloride, chloroform or tetracol phenixin.
Work as R 2Be OH or OCOR 6R 3Be CH 2OH, CH 2OCOR 6R 4Be CONHR 6COOCH 3, CONHR 6COOH, CONHR 6CONHR 6COOR 6Or CONHR 6CONHR 6During COOH, preparation process comprises:
A. amino acid ester or amino acid ester hydrochloride are dissolved in the halogenated organic solvent, add 3,23-O-diacyl-28-white birch acid acyl-oxygen reacted 1-48 hour, obtained 3,23-O-diacyl-28-white birch acid acyl ammonia amino acid ester;
B. a product is dissolved in the solution of halogenated organic solvent or THF (tetrahydrofuran (THF)) and methyl alcohol, adds the strong base solution of 1-8mol/L, back flow reaction 1-8 hour, obtain 3,23-dihydroxyl-28-white birch acid amidoamino acid;
C. condensing agent, organic bases and amino acid ester or amino acid ester hydrochloride and above-mentioned product are dissolved in the halogenated solvent, reacted 1-48 hour, obtain 3,23-dihydroxyl-28-white birch acid amidoamino acid amidoamino acid ester;
D. the c product is dissolved in the solution of halogenated organic solvent or ether and methyl alcohol, adds the strong base solution of 1-8mol/L, back flow reaction 1-8 hour, obtain 3,23-dihydroxyl-28-white birch acid amidoamino acid amidoamino acid.
Wherein halogenated solvent is methylene dichloride, chloroform or tetracol phenixin; Strong base solution is sodium hydroxide or potassium hydroxide solution; Condensing agent is N, N-dicyclohexylcarbodiimide, N-(3-dimethyl amine propyl group)-N '-ethyl carbodiimide; Organic bases is 4-dimethyl amine pyridine, 1-hydroxyl-benzo-triazole or N-methylmorpholine.
Work as R 2Be OH or OCOR 6R 3Be CH 2OH or CH 2OCOR 6R 4Be COOR 6R 5Be CH 2=C-CH 2During OH, preparation process comprises:
A. with 3,23-O-acyl group-28-carboxylicesters betulinic acid derivative and metachloroperbenzoic acid are dissolved in the halogenated solvent, back flow reaction 4-10 hour, obtain 3,23-O-diacyl-30-hydroxyl-28-carboxylicesters white birch acid;
B. a product is dissolved in the solution of halogenated organic solvent or THF (tetrahydrofuran (THF)) and methyl alcohol, adds the strong base solution of 1-8mol/L, back flow reaction 1-8 hour, obtain 3,23,30-trihydroxy--17-carboxylicesters white birch acid;
Wherein halogenated solvent is methylene dichloride, chloroform or tetracol phenixin; Strong base solution is sodium hydroxide or potassium hydroxide solution.
Work as R 2Be OH, OCOR, OCOR 6COOH ,=O, NOH or NOCOR 6R 3During for OH, preparation process comprises:
The protected 23-hydroxyl radical white birck acid of a, the acid of 23-OH white birch or 28-COOH is protected with selective protection reagent (can be selected from triphenylmethyl chloride, dimethyl tertiary butyl chloride silane), or directly uses;
B, an amount of acid anhydrides or acyl chlorides or oxidizer treatment 23-hydroxyl radical white birck acid or derivatives thereof 4-48 hour, temperature was a room temperature to 120 ℃;
C, in the solution of step b, add entry;
D, use organic solvent extraction, obtain organic layer, described derivative is present in the organic layer;
E, the d product is sloughed blocking group in formic acid solution, obtain 3-O-acyl group-23-hydroxyl-28-white birch acid or 3-ketone-23-hydroxyl-28-white birch acid;
F, with e product 3-ketone-23-hydroxyl-28-white birch acid esters, oxammonium hydrochloride is dissolved in the pyridine, room temperature to 70 ℃ is stirred to and reacts completely;
G, with f product organic solvent extraction, obtain organic layer, described 3-oximido-23-hydroxyl radical white birck acid derivative is present in the organic layer;
Wherein oxygenant is PCC oxygenant (pyridinium chlorochromate) or Jone ' s oxygenant (H 2CrO 4/ H 2SO 4/ acetone); Organic solvent is methylene dichloride, chloroform or ethyl acetate.
Part derivative of the present invention can represent that preparation process is (as not clearly marked, then R with the following formula reaction scheme 2, R 3, R 4, R 6Definition before described):
Reaction scheme 1:
Can R be arranged the synthetic type of compounds by reaction scheme 1 2Represent OH, OCOR 6R 3Represent CH 2OH, CH 2OCOR 6R 4Represent COOH, CONHR 6, COOR 6, COOR 6OH, COO (CH 2) 4OCO R 6, COOR 6OCOR 6COOH, CONHR 6COOCH 3Or CONHR 6COOH.
Reaction scheme 2:
Figure C20061004027700103
Can R be arranged the synthetic type of compounds by reaction scheme 2 2Represent OCOR 6R 3Represent CH 2OCOR 6R 4Represent COOR 6OCOR 6COOH or CONHR 6NHCOR 6COOH.
Reaction scheme 3:
Figure C20061004027700111
EDC is N-(3-dimethyl amine propyl group)-N '-ethyl carbodiimide, and HOBT is 1-hydroxyl-benzo-triazole.
By reaction scheme 3 main synthetic types of compounds is R 2And R 3Represent OH, R 4Represent CONHR 6CONHR 6COOR 6Or CONHR 6CONHR 6COOH.
Reaction scheme 4:
Figure C20061004027700112
Wherein m-CPBA is a metachloroperbenzoic acid
By reaction scheme 4 main synthetic types of compounds is R 2Represent OH, OCOR 6R 3Representative, CH 2OH, CH 2OCOR 6R 4Represent COO R 6R 5Represent CH 2=C-CH 2OH.
Reaction scheme 5:
Figure C20061004027700121
TrCl represents triphenylmethyl chloride, and DMAP is the 4-dimethylamino pyridine
Type by reaction scheme 5 main synthetic compounds has R 2Represent OH, OCOR 6R 3Represent CH 2OH, CH 2OCOR 6R 4Represent COOH.
Reaction scheme 6:
Figure C20061004027700122
By reaction scheme 6 main synthetic types of compounds R is arranged 2Represent OCOCO OR 6R 3Represent CH 2OCOCO OR 6R 4Represent COO R 6
Reaction scheme 7
Figure C20061004027700131
TBSCl is a TERT-BUTYL DIMETHYL CHLORO SILANE, and PCC is a pyridinium chlorochromate PCC oxygenant, and DMAP is a 4-dimethyl amine pyridine.
Mainly contain R by reaction scheme 7 main synthetic types of compounds 2Representative=O, NOH or NOCOR 6R 3Represent CH 2OH, CH 2OCOOR 6R 4Represent COOR 6
Reaction scheme 8
Figure C20061004027700132
PCC is a pyridinium chlorochromate PCC oxygenant, and Jone ' s oxygenant is H 2CrO 4/ H 2SO 4/ acetone.
By reaction scheme 8 available types of compounds R is arranged 2Representative=O; R 3Represent CHO or COOH.
The structure of synthetic part of compounds of the present invention sees Table 1:
Table 1 part of compounds structure (R 1Be H)
Figure C20061004027700141
Figure C20061004027700151
23-hydroxyl radical white birck acid derivative of the present invention can form pharmacy acceptable salt with alkali.Described alkali can be sodium hydroxide, potassium hydroxide, lithium hydroxide etc.
Pharmacological testing proves that 23-hydroxyl radical white birck acid derivative of the present invention has antitumor action, can be used to prepare antitumor drug.The tumor disease of preferred therapeutic is liver cancer, lung cancer, cancer of the stomach, melanoma, cervical cancer, mammary cancer, leukemia or brain tumor.
Be the pharmacology test and the data of part of compounds of the present invention below:
One, external MTT screening method
1 gets and is in one bottle in cell in good condition exponential phase of growth, adds 0.25% tryptic digestive juice, and digestion comes off attached cell, counting 2～4 * 10 4Individual/ml, make cell suspension.
2 obtained cell suspensions are inoculated on 96 orifice plates, and constant temperature CO is put in 180 μ l/ holes 2Cultivated 24 hours in the incubator.
3 change liquid, add to be subjected to the reagent thing, and cultivated 48 hours in 20 μ l/ holes.
4 add MTT in 96 orifice plates, 20 μ l/ holes, and reaction is 4 hours in the incubator.
Supernatant liquor is removed in 5 suctions, adds DMSO, 150 μ l/ holes, and jolting is 5 minutes on the dull and stereotyped shaking table.
6 usefulness enzyme-linked immunosorbent assay instruments are the light absorption value that the 570nm place measures every hole at wavelength, and calculate cell inhibitory rate
Figure C20061004027700171
Table 2
Figure C20061004027700172
Table 3
Figure C20061004027700181
Table 4
Figure C20061004027700182
Table 5
Figure C20061004027700183
Table 6
Figure C20061004027700184
Table 7
Figure C20061004027700191
Table 8
Figure C20061004027700201
Annotate: BA is white birch acid in table 1～table 8, and 23HBA is the 23-hydroxyl radical white birck acid.
The clone code table is shown:
BGC-823 represents that the low differentiation of people adenocarcinoma of stomach cell BEL-7402 represents human liver cancer cell
A549 represents that human lung carcinoma cell SF-763 represents the human brain oncocyte
SMMC-7721 represents that human liver cancer cell MDA-MB-231 represents human breast cancer cell
HL-60 represents that human promyelocytic leukemia cell HO-8910 represents Proliferation of Human Ovarian Cell
B16 represents that mouse melanin tumor cell HELA represents human cervical carcinoma cell
Above-mentioned test shows that part of compounds of the present invention all has certain restraining effect for different tumours, and effect is better than the 23-hydroxyl radical white birck acid.
Two, anti-tumor in vivo test
Compd B Y-59 is to the experimental therapy effect of mouse H22 liver cancer
Select for use 18-22 to restrain the H22 knurl kind of female ICR mouse and well-grown 7-11 days, it is subcutaneous to be inoculated in the right side of mice armpit, about 4.5-5 * 10 6Cell/only, inoculate and divide cage at random, intraperitoneal administration after 24 hours.Stop giving 2 days again in 2 days by various dose successive administration 4 days or to 2 days.Put to death animal on the 8th day, weigh, knurl is heavy, calculate that respectively to organize average knurl heavy, press formula and obtain tumor control rate and carry out the T check.
Figure C20061004027700202
The results are shown in Table 9.Table 9 shows: compd B Y-5920mg/kg is to the high greatly and 23-hydroxyl radical white birck acid of tumor growth restraining effect of mouse H22 liver cancer.
Table 9
Annotate: 23HBA is the 23-hydroxyl radical white birck acid
Three, anti-tumor in vivo test
Compd B Y-59 is to the experimental therapy effect of mouse B16 sarcoma
Select for use 18-22 to restrain the B16 knurl kind of female ICR mouse and well-grown 7-11 days, it is subcutaneous to be inoculated in the right side of mice armpit, about 4.5-5 * 10 6Cell/only, inoculate and divide cage at random, intraperitoneal administration after 24 hours.Pressed same dosage successive administration 11 days.Put to death animal on the 12nd day, weigh, knurl is heavy, calculate that respectively to organize average knurl heavy, press formula and obtain tumor control rate and carry out the T check.
Figure C20061004027700211
The results are shown in Table 10.Table 10 shows: compd B Y-5920mg/kg of the present invention is much higher than the 23-hydroxyl radical white birck acid to the tumor growth restraining effect of mouse B16 sarcoma.
Table 10
Figure C20061004027700212
Annotate: 23HBA is the 23-hydroxyl radical white birck acid
The invention also discloses a kind of pharmaceutical composition, wherein contain general formula (I) compound and the pharmaceutically acceptable carrier for the treatment of significant quantity.Described pharmaceutical composition can be a dosage form conventional on the technology of pharmaceutics such as tablet, capsule, injection.Usually dosage be 1.0-400mg/ people/time, also can depart from this dosage range according to the severity difference of disease and the difference of formulation.
Embodiment
Embodiment 1
3,23-O-diacetyl white birch acid (X3)
With the 5ml pyridine with the 23-hydroxyl radical white birck acid (0.12g 0.25mmol) is dissolved in the 10ml eggplant-shape bottle, drip acetic anhydride (0.15ml, 1.5mmol), stirred overnight at room temperature.The dilution of 10ml ethyl acetate, 9% salt acid elution pH to 4-5, saturated aqueous common salt (5ml * 3) washing, anhydrous magnesium sulfate drying filters, and re-crystallizing in ethyl acetate gets pale yellow powder shape solid 0.136g, yield 98%.
ESI-MS：579[M+Na] +
1HNMR(300Hz，CDCl 3)，δ(ppm)：0.80(s，3H，CH 3)，0.88(s，3H，CH 3)，0.93(s，3H，CH 3)，0.97(s，3H，CH 3)，1.69(s，3H，CH 3)，2.01(s，3H，CH 3)，2.06(s，3H，CH 3)，3.00(m，1H，C 19-H)，3.68～3.70(d，1H，23-H)，3.83～3.85(d，1H，23-H)，4.61(s，1H，29-H)，4.74(s，1H，29-H)，4.75～4.78(m，1H，3-H)
Embodiment 2
3,23-O-diacetyl-28-white birch acyl chlorides (BYA-2)
With 3, (0.1g 0.18mmol) is dissolved in the 10ml eggplant-shape bottle 23-O-diacetyl-28-acid-white birch acid, drips oxalyl chloride 0.1ml, and stirring at room 4h boils off methylene dichloride with the refining exsiccant methylene dichloride of 5ml.Add refining exsiccant methylene dichloride (5ml * 3) again, dissolving, steaming removes.Obtain pale-yellow galss shape solid 0.1g at last.
Embodiment 3
3,23-diacetyl-28-white birch acid methyl esters (SZ1)
(0.1g 0.17mmol) is dissolved in the 10ml eggplant-shape bottle BYA-2 that embodiment 2 methods is prepared with the refining exsiccant methylene dichloride of 5ml, drips methyl alcohol 5ml, stirring at room 6h.Saturated aqueous common salt (5ml * 3) washing, anhydrous sodium sulfate drying filters, and filtrate concentrates system sand.Column chromatography [sherwood oil (60 ℃～90 ℃): ethyl acetate=12: 1 (v: v)], get white powder solid 0.092g, total recovery 90.0%, 110～112 ℃ of mp.
ESI-MS：593[M+Na] +
1HNMR(300Hz，CDCl 3)，δ(ppm)：0.78(s，3H，CH 3)，0.83(s，3H，CH 3)，0.87(s，3H，CH 3)，0.89(s，3H，CH 3)，1.68(s，3H，CH 3)，1.99(s，3H，CH 3)，2.04(s，3H，CH 3)，2.94～3.01(m，1H，C 19-H)，3.64(s，3H，CH 3-O)，3.59～3.68(d，1H，23-H)，3.79～3.83(d，1H，23-H)，4.57(s，1H，29-H)，4.71(s，1H，29-H)，4.71～4.77(m，1H，3-H)
Embodiment 4
3, the positive butyramide white birch of 23-diacetyl-28-acid (XA1)
(0.1g 0.17mmol) is dissolved among the exsiccant methylene dichloride 5ml BYA-2 that embodiment 2 methods are prepared, and drips n-Butyl Amine 99 3ml, drips the 0.05ml triethylamine again, stirring at room 7h.9% salt acid elution pH to 4-5, saturated aqueous common salt (5ml * 3) washing, anhydrous sodium sulfate drying filters, and filtrate concentrates system sand.Column chromatography [methylene dichloride: acetone=60: 1 (v: v)], get white powder solid 0.093g, total recovery 85.0%, 115～117 ℃ of mp.
ESI-MS：612[M+H] +
1HNMR(300Hz，CDCl 3)，δ(ppm)：0.77(s，3H，CH 3)，0.85(s，3H，CH 3)，0.90～0.93(m，9H，3CH 3)，1.65(s，3H，CH 3)，1.99(s，3H，CH 3)，2.04(s，3H，CH 3)，3.13～3.15(m，1H，C 19-H)，3.25～3.32(m，2H，CH 2-N)，3.63～3.66(d，1H，23-H)，3.80～3.84(d，1H，23-H)，4.56(s，1H，29-H)，4.70(s，1H，29-H)，4.70～4.77(m，1H，3-H)，5.52～5.56(t，1H，H-N)
Embodiment 5
3,23-O-diacetyl-28-white birch acid acid amides glycine methyl ester (BY-19-3)
(0.07g, 0.54mmol), (0.012g 0.1mmol) is dissolved in the 10ml exsiccant methylene dichloride DMAP, drips BYA-2 (0.1g, dichloromethane solution 0.17mmol), the stirring at room 12h of embodiment 2 with glycine methyl ester hydrochloride.9% salt acid elution pH to 7, saturated aqueous common salt (5ml * 3) washing, Na 2SO 4Drying is filtered, and filtrate concentrates system sand.Column chromatography [sherwood oil: acetone=5: 1 (v: v)], get white solid 0.062g, total recovery 55%, mp109-111 ℃.
ESI-MS：628.3[M+H] +
1HNMR (300Hz, CDCl 3), δ (ppm): 0.80 (s, 3H, CH 3), 0.87 (s, 3H, CH 3), 0.91 (s, 3H, CH 3), 0.96 (s, 3H, CH 3), 1.58 (s, 3H, CH 3), 2.01,2.06 (s, each 3H, 3,23-COOCH 3), 3.09 (m, 1H, C 19-H), 3.76 (s, 3H, COOCH 3), 3.76 (dd, 2H, 23-H), 4.00 (t, 2H, N-CH 2), 4.66 (d, 2H, 29-H), 4.75 (m, 1H, 3-H), 6.05 (t, 1H, CONH)
Embodiment 6
3,23-O-diacetyl-30-hydroxyl-28-white birch acetoacetic ester (BYA-4-3)
With 3, (0.1g, 0.17mmol), (0.03g 0.18mmol) is dissolved in the 10ml chloroform m-CPBA 23-O-diacetyl-28-white birch acetoacetic ester, and heating reflux reaction 6h uses saturated NaHCO 3(10ml*2) washing, and then use the 10ml water washing, with saturated aqueous common salt 10ml washing, anhydrous sodium sulfate drying filters at last, and filtrate concentrates system sand.Column chromatography [sherwood oil: ethyl acetate=3: 1 (v: v)], get white solid 0.05g, total recovery 47%, mp110-111 ℃.
ESI-MS：623.5[M+Na] +
1HNMR (300Hz, CDCl 3), δ (ppm): 0.80 (s, 3H, CH 3), 0.87 (s, 3H, CH 3), 0.93 (s, 3H, CH 3), 0.96 (s, 3H, CH 3), 1.58 (s, 3H, CH 3), 2.01,2.06 (s, each 3H, 3,23-COOCH 3), 2.89 (m, 1H, C 19-H), 3.76 (dd, 2H, 23-H), 4.12 (m, 4H, 30-CH 2With 28-COO CH 2), 4.76 (m, 1H, 3-H), 4.94 (d, 2H, 29-H)
Embodiment 7
3,23-dihydroxyl-28-white birch acid n-pentyl ester (BY-61)
With 3; (0.1g 0.16mmol) is dissolved in the mixing solutions 6ml (v/v=1: 1), add the NaOH solution 2ml of 4M of THF and methyl alcohol to 23-O-diacetyl-28-white birch acid n-pentyl ester; heating reflux reaction 4 hours; transfer PH to 7 with 10%HCl, have solid to separate out, filter; obtain white solid 0.08g;, productive rate 95%, mp76-78 ℃.
ESI-MS：543.4[M+H] +
1HNMR(300Hz，DMSO-D6)，δ(ppm)：0.51(s，3H，CH 3)，0.77(s，3H，CH 3)，0.84(s，3H，CH 3)，0.93(s，3H，CH 3)，1.65(s，3H，CH 3)，2.90(m，1H，C 19-H)，3.76(s，3H，COOCH 3)，3.18(dd，2H，23-H)，3.45(m，1H，3-H)，4.02(m，2H，28-COOCH 2)，4.10(d，1H，3-OH)，4.57(t，1H，23-OH)，4.60(d，2H，29-H)
Embodiment 8
3,23-dihydroxyl-28-benzamide white birch acid (I1)
With 3, (0.13g 0.20mmol) is dissolved in the CH of 3mlTHF and 1.5ml to 23-O-diacetyl-28-benzamide white birch acid 3Among the OH, to the NaOH aqueous solution (1mol/L) that wherein drips 1.5ml, the reaction solution stirred overnight at room temperature, concentrating under reduced pressure is removed organic solvent, adds the ethyl acetate dilution of 50ml.Organic phase is washed to water layer with 5% aqueous hydrochloric acid and is shown slightly acidic, and water is given a baby a bath on the third day after its birth time again, saturated common salt washing twice, anhydrous Na 2SO 4Dry two hours.Be evaporated to driedly, silica gel column chromatography separates, sherwood oil: and ethyl acetate=2: 3 (v: v) wash-out, obtain white solid 0.10g, productive rate is 95%, mp 144-146 ℃.
ESI-MS：528[M+H] +
1H-NMR(CDCl 3，300MHz)：δ(ppm)：0.85(3H，s，CH 3)，0.86(3H，s，CH 3)，0.94(3H，s，CH 3)，0.96(3H，s，CH 3)，1.67(3H，s，CH 3)，3.09～3.19(2H，m，N-CH 2)，3.26～3.32(1H，m，C 19-H)，3.40(1H，d，23-H)，3.58～3.63(1H，m，3-H)，3.70(1H，d，23-H)，4.58and?4.72(1H?each，brs，29-H)，5.57(1H，t，N-H).
Embodiment 9
3,23,30-trihydroxy--28-white birch acetoacetic ester (BY-65)
With 3, (0.05g 0.083mmol) is dissolved in the CH of 3mlTHF and 1.5ml to 23-O-diacetyl-30-hydroxyl-28-white birch acetoacetic ester 3Among the OH, to the NaOH aqueous solution (4mol/L) that wherein drips 1.5ml, back flow reaction 5h, thin up is transferred PH to 7, has solid to separate out, and filters, and obtains white solid 0.04g, and productive rate is 95%, mp 248-250 ℃ (dec)
ESI-MS：539.3[M+Na] +
1HNMR (300Hz, CDCl 3), δ (ppm): 0.86 (s, 3H, CH 3), 0.87 (s, 3H, CH 3), 0.91 (s, 3H, CH 3), 0.96 (s, 3H, CH 3), 1.25 (s, 3H, CH 3), 2.89 (m, 1H, C 19-H), 3.56 (dd, 2H, 23-H), 3.61 (m, 1H, 3-H), 4.13 (m, 4H, 30-CH 2With 28-COO CH 2), 4.93 (d, 2H, 29-H)
Embodiment 10
3,23-dihydroxyl-28-white birch acid acid amides glycine (BY-26)
With 3, (0.06g 0.096mmol) is dissolved in the CH of 3mlTHF and 1.5ml to 23-O-diacetyl-28-white birch acid acid amides glycine methyl ester 3Among the OH, to the NaOH aqueous solution (4mol/L) that wherein drips 1.5ml, back flow reaction 5h, thin up is transferred PH to 4-5, and the adularescent solid is separated out, and filters, and obtains white solid 0.048g, productive rate is 95%, mp 213-215 ℃ ESI-MS:528.4[M-H] -
1HNMR (300Hz, CDCl 3), δ (ppm): 0.52 (s, 3H, CH 3), 0.77 (s, 3H, CH 3), 0.83 (s, 3H, CH 3), 0.91 (s, 3H, CH 3), 1.62 (s, 3H, CH 3), 2.90 (m, 1H, C 19-H), 3.20 (dd, 2H, 23-H), 3.29-3.73 (m, 4H, 3-H and 3-OH, 23-OH, 28-NCH 2), 4.13 (m, 4H, 30-CH 2With 28-COO CH 2), 4.59 (d, 2H, 29-H), 7.86 (t, 1H, 28-CONH)
Embodiment 11
3,23-dihydroxyl-28-white birch acid acid amides glycine acid amides glycine methyl ester (BY-63-3)
With 3, the 23-dihydroxyl-(0.1g 0.19mmol) is dissolved in the 10ml methylene dichloride 28-white birch acid acid amides glycine, adds EDC (0.06g, 0.38mmol), HOBT (0.06g, 0.48mmol), glycine methyl ester hydrochloride (0.035g, 0.29mmol), room temperature reaction spends the night, and filters, and filtrate concentrates system sand.Column chromatography [sherwood oil: acetone=5: 1 (v: v)], get white solid 0.09g, total recovery 80%, mp213-215 ℃.
ESI-MS：601.3[M+H] +
1HNMR(300Hz，CDCl 3)，δ(ppm)：0.85(s，3H，CH 3)，0.88(s，3H，CH 3)，0.91(s，3H，CH 3)，0.96(s，3H，CH 3)，1.68(s，3H，CH 3)，3.05(m，1H，C 19-H)，3.55(dd，2H，23-H)，3.61(m，1H，3-H)，3.75(s，1H，COOCH 3)，3.94-4.06(m，4H，28-NCH 2*2)，4.66(d，2H，29-H)，6.33(t，1H，28-CONH)，6.66(t，1H，28-CONH)
Embodiment 12
3,23-O-diacetyl-28-white birch acid ethylene glycol ester (BY-8)
With 3,23-O-diacetyl-28-white birch acyl chlorides is dissolved in the 10ml eggplant-shape bottle with the refining exsiccant methylene dichloride of 5ml, adds big excessive ethylene glycol, stirring at room 6h.Saturated aqueous common salt (5ml * 3) washing, anhydrous sodium sulfate drying filters, and filtrate concentrates system sand.Column chromatography [sherwood oil (60 ℃～90 ℃): ethyl acetate=3: 1 (v: v)], get white solid 0.07g, total recovery 70.0%, mp 85-87 ℃.
ESI-MS：601.6[M+H] +
1HNMR(300Hz，CDCl 3)，δ(ppm)：0.80(s，3H，CH 3)，0.87(s，3H，CH 3)，0.93(s，3H，CH 3)，0.96(s，3H，CH 3)，1.68(s，3H，CH 3)，2.01(s，3H，CH 3)，2.06(s，3H，CH 3)，2.98(m，1H，C 19-H)，3.76(dd，2H，23-H)，3.83(m，2H，28-C H 2 OH)，4.23(m，2H，28-COOCH 2)，4.66(d，2H，29-H)，4.74～4.77(m，1H，3-H)
Embodiment 13
3,23-O-diacetyl-28-white birch acid ethylene glycol succsinic acid dibasic acid esters (BY-44)
With 3, and 23-O-diacetyl-28-white birch acid esters ethyl alcohol (0.07g .0.12mmol); Succinic anhydried (0.05g, 0.5mmol), DMAP (0.012g; 0.1mmol) be dissolved in the 10ml methylene dichloride; room temperature reaction 16h, with 10%HCl (2*10ml), water 10ml washing; use the water washing of 10ml saturated common salt at last; anhydrous sodium sulfate drying filters, and filtrate concentrates system sand.Column chromatography [sherwood oil (60 ℃～90 ℃): ethyl acetate: acetate=50: 50: 0.4 (v: v)], get faint yellow solid 0.057g, total recovery 70.0%, mp 78-80 ℃.
ESI-MS：699.3[M-H] -
1HNMR(300Hz，CDCl 3)，δ(ppm)：0.81(s，3H，CH 3)，0.87(s，3H，CH 3)，0.91(s，3H，CH 3)，0.96(s，3H，CH 3)，1.68(s，3H，CH 3)，2.01(s，3H，CH 3)，2.06(s，3H，CH 3)，2.66(m，4H，28-OCOC H 2 C H 2 COO)，3.00(m，1H，C 19-H)，3.76(dd，2H，23-H)，3.75(m，2H，28-C H 2 OH)，4.29(m，4H，28-COOC H 2 C H 2 O)，4.66(d，2H，29-H)，4.73(m，1H，3-H)
Embodiment 14
3,23-O-disuccinic acid monoacyl white birch acid (BY-24-2)
(0.1g, 0.21mmol), (0.12g, 1.2mmol), (0.05g 0.4mmol) is dissolved in the 10ml pyridine DMAP Succinic anhydried, is heated to 90 ℃ with the 23-hydroxyl radical white birck acid.Reaction 48h, reaction solution is poured in the cold water, transfer PH to 4-5 with 10%HCl, have solid to separate out, filter, filter cake is husky with methylene dichloride dissolving system, column chromatography [sherwood oil (60 ℃～90 ℃): ethyl acetate: acetate=50: 50: 0.4 (v: v)], get faint yellow solid 0.09g, total recovery 75%, mp 153-155 ℃.
ESI-MS：671.3[M-H] -
1HNMR(300Hz，DMSO-D6)，δ(ppm)：0.75(s，3H，CH 3)，0.83(s，3H，CH 3)，0.86(s，3H，CH 3)，0.94(s，3H，CH 3)，1.64(s，3H，CH 3)，2.45(m，8H，3，23-COC H 2 C H 2 CO)，2.90(m，1H，C 19-H)，3.70(dd，2H，23-H)，4.62(d，2H，29-H)，4.67(m，1H，3-H)，12.10(s，3H，3，23，28-COOH)
Embodiment 15
3-hydroxyl-23-O-triphenyl methyl ether white birch acid (BY-34-1)
(0.1g, 0.21mmol), (0.12g, 0.42mmol), (0.014g 0.12mmol), is dissolved in the 10ml pyridine DMAP tri-phenyl chloride, is heated to 90 ℃ with the 23-hydroxyl radical white birck acid.Reaction 24h, reaction solution is poured in the cold water, transfers PH to 4-5 with 10%HCl, has solid to separate out, filter, filter cake is husky with methylene dichloride dissolving system, column chromatography [sherwood oil (60 ℃～90 ℃): ethyl acetate=3: 1 (v: v)], get pale solid 0.10g, total recovery 65%, mp200-202 ℃.
ESI-MS：737[M+Na] +
1HNMR(300Hz，CDCl 3)，δ(ppm)：0.71(s，3H，CH 3)，0.82(s，3H，CH 3)，0.89(s，3H，CH 3)，1.00(s，3H，CH 3)，1.69(s，3H，CH 3)，2.99～3.00(m，1H，C 19-H)，2.77～2.80(d，1H，23-H)，3.18～3.21(d，1H，23-H)，3.59～3.65(m，1H，3-H)，4.61(s，1H，29-H)，4.74(s，1H，29-H)，7.23～7.46(m，15H，3ArH)
Embodiment 16
3-O-phthalic acid monoacyl-23-O-triphenyl methyl ether white birch acid (BY-30-3)
(0.1g, 0.14mmol), (0.04g, 0.28mmol), (0.012g 0.1mmol), is dissolved in the 10ml pyridine DMAP Tetra hydro Phthalic anhydride, is heated to 90 ℃ with 3-hydroxyl-23-O-triphenyl methyl ether-28-white birch acid.Reaction 24h, reaction solution is poured in the cold water, transfers PH to 4-5 with 10%HCl, have solid to separate out, filter, filter cake is husky with methylene dichloride dissolving system, column chromatography [sherwood oil (60 ℃～90 ℃): ethyl acetate: acetate=50: 50: 0.4 (v: v)], get pale solid 0.07g, total recovery 60%.
ESI-MS：885.5[M+Na] +
Embodiment 17
3-O-phthalic acid monoacyl-23-hydroxyl radical white birck acid (BY-30-4)
(0.07g 0.08mmol) is dissolved in the 10ml chloroform, adds 2ml formic acid with 3-O-(2-acid+benzoic acid ester)-23-O-triphenyl methyl ether-28-white birch acid, back flow reaction 4h, evaporate to dryness adds water, has solid to separate out, filter, filter cake is husky with methylene dichloride dissolving system, column chromatography [sherwood oil (60 ℃～90 ℃): ethyl acetate: acetate=50: 50: 0.4 (v: v)], get yellow solid 0.035g, total recovery 65%, 280 ℃ of mp (dec).
ESI-MS：647.4[M-H] -
1HNMR(300Hz，DMSO-D6)，δ(ppm)：0.74(s，3H，CH 3)，0.84(s，3H，CH 3)，0.89(s，3H，CH 3)，0.96(s，3H，CH 3)，1.65(s，3H，CH 3)，2.95(m，1H，C 19-H)，3.91(d，1H，23-H)，4.24(d，1H，23-H)，4.56(s，1H，29-H)，4.69(s，1H，29-H)，4.82(m，1H，3-H)，7.23～7.61(m，4H，ArH)
Embodiment 18
3-O-benzoic ether-23-triphenyl methyl ether white birch acid (LD-34)
With the 10ml pyridine with 3-hydroxyl-23-O-triphenyl methyl ether-28-white birch acid (0.1g 0.14mmol) is dissolved in the 25ml round-bottomed flask, add Benzoyl chloride (0.1ml, 0.86mmol), add again DMAP (0.012g, 0.1mmol), stirring at room 2h.The dilution of 20ml ethyl acetate, 9% salt acid elution pH to 4-5, saturated aqueous common salt (10ml * 3) washing, anhydrous magnesium sulfate drying filters, and filtrate concentrates system sand.Column chromatography [sherwood oil (60 ℃～90 ℃): ethyl acetate=10: 1 (v: v)], get white powder solid 0.08g, yield 70%
ESI-MS：841[M+Na] +
Embodiment 19
3-hydroxyl-23-O-benzoic ether white birch acid (XB3a)
(0.08g 0.097mmol) is dissolved in the 10ml exsiccant methylene dichloride, drips 1ml formic acid, stirring at room 30min with 3-O-benzoic ether-23-triphenyl methyl ether-28-white birch acid.Saturated NaHCO 3Solution washing pH to 7-8, saturated aqueous common salt (10ml * 3) washing, anhydrous magnesium sulfate drying filters, and filtrate concentrates system sand.Column chromatography [sherwood oil (60 ℃～90 ℃): ethyl acetate=8: 1 (v: v)], get white powder solid 0.033g, yield 55.6%, 144～145 ℃ of mp.
ESI-MS：575[M-H] -
1HNMR(300Hz，CDCl 3)，δ(ppm)：0.88(s，3H，CH 3)，0.89(s，3H，CH 3)，1.02(s，6H，2CH 3)，1.72(s，3H，CH 3)，3.03～3.09(m，1H，C 19-H)，3.42～3.50(d，1H，23-H)，3.62～3.65(t，1H，3-H)，3.73～3.74(d，1H，23-H)，4.65(s，1H，29-H)，4.77(s，1H，29-H)，7.49～8.05(m，5H，ArH)
Embodiment 20
3-ketone-23-aldehyde white birch acid (Y1)
With the 23-hydroxyl radical white birck acid (0.12g 0.25mmol) is dissolved among the 10ml methylene dichloride 25ml, add the PCC be stated from aluminum oxide (0.66g, 0.5mmol), stirring at room.Behind the 15min, reaction solution is become brown by yellowish-orange.Continue to stir 5.5h, stopped reaction, inclining supernatant liquor.Add anhydrous diethyl ether 10ml, fully stir 15min, inclining supernatant liquor.Handle again twice with anhydrous diethyl ether with same method.Merge organic layer, the silica gel short column of flowing through concentrates system sand.Column chromatography [methylene dichloride: acetone=100: 1 (v: v)], get white powder solid 0.082g, yield 70.5%, 210～212 ℃ of mp.
1HNMR(300Hz，CDCl 3)，δ(ppm)：0.97～1.25(m，9H，3CH 3)，1.27(s，3H，CH 3)，1.70(s，3H，CH 3)，2.99～3.03(m，1H，C 19-H)，4.62(s，1H，29-H)，4.75(s，1H，29-H)，9.34(s，1H，H-C＝O)Anal.C 30H 44O 4
Found(％)：C?76.32 H?9.74
Calcd(％)：C?76.88 H?9.46
Embodiment 21
3-ketone-23-acid white birch acid (Y2)
(0.12g 0.25mmol) is dissolved among the 10ml acetone 25ml, drips the Jones oxygenant of prepared fresh, no longer becomes green in the short period of time up to reaction solution, adds the oxygenant of one times of amount again, and ice bath stirs 1.5h down with the 23-hydroxyl radical white birck acid.Add 1ml methyl alcohol and fully stir 15min, solution becomes green by tawny.Add 10ml water again, then with the organic solvent evaporate to dryness.Ethyl acetate (10ml * 3) is extracted water layer, merges.The organic layer that saturated aqueous common salt (10ml * 3) washing merges, anhydrous magnesium sulfate drying filters, and filtrate concentrates system sand.Column chromatography [methylene dichloride: acetone=120: 1 (v: v)], get white powder solid 0.073g, yield 60.7%, 218～220 ℃ of mp.
ESI-MS：439[M-COOH] -
1HNMR(300Hz，CDCl 3)，δ(ppm)：0.96～1.02(m，12H，4CH 3)，1.69(s，3H，CH 3)，2.98～3.03(m，1H，C 19-H)，4.61(s，1H，29-H)，4.74(s，1H，29-H)
Embodiment 22
3-hydroxyl-23-O-tertiary butyl dimethyl-silicon ether-28-white birch acid methyl esters (XP-5)
With 3, the 23-dihydroxyl-(0.23g, 0.47mmol), (0.10g 0.82mmol) is dissolved in 10ml exsiccant CH to DMAP to 28-white birch acid methyl esters 2Cl 2In, (0.13g, 0.86mmol), stirred overnight at room temperature adds the excessive TBSCl of 2ml water decomposition, stirs half an hour, adds 40ml CH again to add TERT-BUTYL DIMETHYL CHLORO SILANE (TBSCl) rapidly 2Cl 2Dilute reaction solution.Organic phase use successively 5% aqueous hydrochloric acid (3 * 20ml), water (2 * 20ml) and saturated aqueous common salt (2 * 20ml) washing, anhydrous sodium sulfate drying two hours, concentrating under reduced pressure as for.Silica gel column chromatography separates, sherwood oil: (v: v) wash-out obtains 0.26g white powder solid, productive rate 92%, mp156～158 ℃ to ethyl acetate=16: 1.
ESI-MS：623[M+Na] +
1H-NMR(CDCl 3，300MHz)：δ(ppm)：0.065(3H，s，SiCH 3)，0.068(3H，s，SiCH 3)，0.84(3H，s，CH 3)，0.86(3H，s，CH 3)，0.90(9H，s，3×CH 3)，0.91(3H，s，CH 3)，0.95(3H，s，CH 3)，1.68(3H，s，CH 3)，2.18～2.24(2H，m，2-H 2)，2.97～3.01(1H，m，C 19-H)，3.33(1H，d，23-H)，3.54～3.60(1H，m，3-H)，3.63(1H，d，23-H)，3.66(3H，s，OCH 3)，4.59and?4.73(1H?each，brs，29-H).
Embodiment 22
3-ketone-23-O-tertiary butyl dimethyl-silicon ether-28-white birch acid methyl esters (XP-6)
With 3-hydroxyl-23-O-tertiary butyl dimethyl-silicon ether-(0.22g 0.37mmol) is dissolved in the 10ml exsiccant methylene dichloride, under the room temperature 28-white birch acid methyl esters, add PCC (0.24g, 1.11mmol), stirring 3 hours, black mixture dilutes with the 40ml anhydrous diethyl ether, directly adding 0.5g silica gel room temperature is evaporated to dried, silica gel column chromatography separates, sherwood oil: (v: v) wash-out obtains the white powder solid of 0.14g to ethyl acetate=30: 1, productive rate 64.5%, mp137～139 ℃.
ESI-MS：599[M+H] +
1H-NMR(CDCl 3，300MHz)：δ0.01(6H，s，2×SiCH 3)，0.84(3H，s，CH 3)，0.85(3H，s，CH 3)，0.87(9H，s，3×CH 3)，0.96(3H，s，CH 3)，0.98(3H，s，CH 3)，1.69(3H，s，CH 3)，2.97～3.01(1H，m，C 19-H)，3.29(1H，d，J＝9.2Hz，23-H)，3.57(1H，d，J＝9.2Hz，23-H)，3.67(3H，s，OCH 3)，4.60and?4.74(1Heach，brs，29-H).
Embodiment 23
3-ketone-23-hydroxyl-28-white birch acid methyl esters (II)
With 3-ketone-23-O-tertiary butyl dimethyl-silicon ether-28-white birch acid methyl esters (0.16g, 0.27mmol) be dissolved among the THF of 5ml, to the THF solution (0.5mol/L) that wherein drips the 1ml tetrabutyl ammonium fluoride, stirring at room 3 hours is evaporated to driedly, and silica gel column chromatography separates, sherwood oil: ethyl acetate=4: 1 (v: v) wash-out, the 0.11g white solid, productive rate 90%, mp186～188 ℃.
ESI-MS：485[M+H] +
1H-NMR(CDCl 3，300MHz)：δ(ppm)0.97(6H，s，2×CH 3)，1.00(3H，s，CH 3)，1.04(3H，s，CH 3)，1.68(3H，s，CH 3)，2.25～2.29(1H，m，2-H)，2.55～2.63(1H，m，2-H)，3.00～3.04(1H，m，C 19-H)，3.40(1H，d，23-H)，3.62(1H，d，23-H)，3.67(3H，s，OCH 3)，4.60and?4.73(1Heach，brs，29-H).
Embodiment 24
3-oximido-23-hydroxyl-28-white birch acid methyl esters (III)
(0.30g, 0.60mmol), (0.3g 4.20mmol) is dissolved in the 10ml pyridine oxammonium hydrochloride, and 50 ℃ were stirred 4 hours, after reaction solution is cooled to room temperature, adds the dilution of 40ml ethyl acetate with 3-ketone-23-OH white birch acid methyl esters.Separate organic layer, and use successively 5% aqueous hydrochloric acid (3 * 20ml), water (2 * 20ml), saturated aqueous common salt (2 * 20ml) washings.The organic layer anhydrous Na 2SO 4After dry two hours, be evaporated to dried.Silica gel column chromatography separates, sherwood oil: ethyl acetate=4: 1 (v: v) wash-out, 0.29g white powder solid, productive rate 95%, mp114～116 ℃.
ESI-MS：500[M+H] +
1H-NMR(CDCl 3，300MHz)：δ(ppm)0.92(6H，s，2×CH 3)，0.95(3H，s，CH 3)，1.02(3H，s，CH 3)，1.68(3H，s，CH 3)，2.96～3.12(2H，m；2-H，C 19-H)，3.49(1H，d，23-H)，3.64(1H，d，23-H)，3.66(3H，s，OCH 3)，4.60and?4.73(1H?each，br?s，29-H).
Embodiment 25
3-ketone-23-ethanoyl-28-white birch acid methyl esters (II1)
With 3-ketone-23-OH white birch acid methyl esters (0.06g, 0.12mmol), DMAP (0.014g, O.12mmol) and diacetyl oxide (0.06g 0.6mmol) is dissolved in 5ml exsiccant CH 2Cl 2In, stirring at room 10h adds 20ml CH 2Cl 2Dilution.The saturated NaHCO of organic phase 3The aqueous solution is washed 3 times, and 5% aqueous hydrochloric acid is washed 2 times, washes saturated common salt washing 1 time, anhydrous Na 2 times 2SO 4Dry 2 hours, be evaporated to dried, column chromatography for separation, sherwood oil: (v: v) wash-out obtains white powder solid 0.062g, productive rate 99%, mp58～60 ℃ to ethyl acetate=10: 1.
ESI-MS：527[M+H] +
1H-NMR(CDCl 3，300MHz)：δ(ppm)O.94(3H，s，CH 3)，0.96(3H，s，CH 3)，0.98(3H，s，CH 3)，0.99(3H，s，CH 3)，1.68(3H，s，CH 3)，2.02(3H，s，COCH 3)，2.42～2.50(2H，m，2-H 2)，2.95～3.04(1H，m，C 19-H)，3.67(3H，s，OCH 3)，4.00～4.10(2H，m，23-H 2)，4.60，4.74(1Heach，brs，29-H).
Embodiment 26
3-ethanoyl oximido-3-O-ethanoyl-28-white birch acid methyl esters (III1)
With 3-oximido-23-hydroxyl-28-white birch acid methyl esters (0.1g, 0.20mmol) and diacetyl oxide (0.2g 2mmol) is dissolved in the anhydrous pyridine of 5ml, and 90 ℃ of oil baths are reacted and spent the night 24 hours reaction times.Be cooled to room temperature, add the dilution of 40ml ethyl acetate.Separate organic layer, use successively then 5% aqueous hydrochloric acid (3 * 20ml), saturated NaHCO 3The aqueous solution (3 * 20ml), water (2 * 20ml), saturated aqueous common salt (2 * 20ml) washing.The organic layer anhydrous Na 2SO 4After dry two hours, be evaporated to driedly, thick product is through column chromatography for separation, sherwood oil: ethyl acetate=6: 1 (v: v) wash-out, white solid 0.11g, productive rate 95%, mp156～160 ℃.
ESI-MS：606[M+Na] +
1H-NMR(CDCl 3，300MHz)：δ(ppm)0.85(3H，s，CH 3)，0.95(3H，s，CH 3)，0.96(3H，s，CH 3)，1.16(3H，s，CH 3)，1.68(3H，s，CH 3)，2.05(3H，s，CH 3-CO)，2.17(3H，s，CH 3-CO)，2.58～2.66(2H，m，2-H 2)，3.00(1H，m，C 19-H)，3.67(3H，s，OCH 3)，4.08(1H，d，23-H)，4.18(1H，d，23-H)，4.61and?4.74(1H?each，br?s，29-H).
Embodiment 27
3,23-dihydroxyl-28-white birch acid propylene ester (BY-59)
(0.1g 0.17mmol) is dissolved in the 10ml eggplant-shape bottle BYA-2 that embodiment 2 methods is prepared with the refining exsiccant methylene dichloride of 5ml, drips vinylcarbinol 2ml, stirring at room 6h.Saturated aqueous common salt (5ml * 3) washing, anhydrous sodium sulfate drying filters, filtrate evaporate to dryness ethyl alcohol recrystallization, filter, filter cake is dissolved in the mixing solutions 8ml (v/v=1: 1), add NaOH solution (4mol/L) 2ml of THF and methyl alcohol, heating reflux reaction 4 hours, thin up is transferred PH to 7 with 10%HCl, has solid to separate out, filter, filter cake concentrates system sand with acetone solution.Column chromatography [sherwood oil (60～90 ℃): ethyl acetate=1: 1 (v: v)], get white powder solid 0.07g, total recovery 75.5%, mp111-112 ℃
ESI-MS：513.3[M+H] +
1HNMR(300Hz，DMSO-D6)，δ(ppm)：0.51(s，3H，CH 3)，0.77(s，3H，CH 3)，0.83(s，3H，CH 3)，0.93(s，3H，CH 3)，1.65(s，3H，CH 3)，2.90(m，1H，C 19-H)，3.18(dd，2H，23-H)，3.45(m，1H，3-H)，4.55(m，2H，28-COOCH 2)，，4.60(d，2H，29-H)，5.27(m，2H，CH＝C H 2 )，5.92(m，1H，C H＝CH 2)
Embodiment 28
Get compound 50mg and starch 500mg that embodiment 27 methods make, dextrin 500mg mixes, and makes wetting agent with an amount of 30% ethanol, makes softwood, and ordinary method is granulated, and adds the 2mg Magnesium Stearate and mixes, and makes tablet.

Claims (4)

1, the 23-hydroxyl radical white birck acid derivative of following arbitrary structure or its pharmacy acceptable salt:
3,23-dihydroxyl-28-white birch acid acid amides glycine;
3,23-dihydroxyl-28-white birch acid acid amides L-Ala;
3,23-dihydroxyl-28-white birch acid acid amides L-glutamic acid;
3,23-dihydroxyl-28-white birch acid acid amides leucine;
3,23-dihydroxyl-28-white birch acid acid amides phenylalanine;
3,23-O-diacetyl-28-white birch acid acid amides glycine methyl ester;
3,23-O-diacetyl-28-white birch acid acid amides glutamic acid dimethyl ester;
3,23-O-diacetyl-28-white birch acid acid amides alanine methyl ester;
3,23-O-diacetyl-28-white birch acid acid amides leucine methyl esters;
3,23-O-diacetyl-28-white birch acid acid amides phenylalanine methyl ester;
3,23-O-diacetyl-28-white birch acid acid amides valine methyl ester;
3, the 23-O-diacetyl-positive butyramide of 28-white birch acid;
3,23-O-diacetyl-28-white birch acid Cyclohexamide;
3,23-O-diacetyl-28-white birch acid DB;
3,23-O-diacetyl-28-white birch acid phenylacetamide;
3,23-O-diacetyl-28-white birch acid diethylamide;
3,23-O-diacetyl-28-white birch acid (3, the 4-(methylenedioxy)) phenylacetamide;
3,23-O-diacetyl-28-white birch acid (Alpha-Methyl) benzyl acid amides; Or
3,23-dihydroxyl-28-white birch acid acid amides glycine acid amides glycine methyl ester.
2, pharmaceutical composition wherein contains the 23-hydroxyl radical white birck acid derivative and the pharmaceutically acceptable carrier of the claim 1 for the treatment of significant quantity.
3, the 23-hydroxyl radical white birck acid derivative of claim 1 is used to prepare the purposes of the medicine for the treatment of tumor disease.
4, the purposes of claim 3, wherein tumor disease is liver cancer, lung cancer, cancer of the stomach, melanoma, cervical cancer, mammary cancer, leukemia or brain tumor.
CNB2006100402772A 2006-05-12 2006-05-12 23-Hydroxyl betulinic acid kind derivant and preparing process, preparation and use thereof Expired - Fee Related CN100475835C (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CNB2006100402772A CN100475835C (en) 2006-05-12 2006-05-12 23-Hydroxyl betulinic acid kind derivant and preparing process, preparation and use thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CNB2006100402772A CN100475835C (en) 2006-05-12 2006-05-12 23-Hydroxyl betulinic acid kind derivant and preparing process, preparation and use thereof

Related Child Applications (1)

Application Number Title Priority Date Filing Date
CN2008100084181A Division CN101230082B (en) 2006-05-12 2006-05-12 23-hydroxy betulinic acid derivative, preparation method, preparation and use thereof

Publications (2)

Publication Number Publication Date
CN1861627A CN1861627A (en) 2006-11-15
CN100475835C true CN100475835C (en) 2009-04-08

Family

ID=37389168

Family Applications (1)

Application Number Title Priority Date Filing Date
CNB2006100402772A Expired - Fee Related CN100475835C (en) 2006-05-12 2006-05-12 23-Hydroxyl betulinic acid kind derivant and preparing process, preparation and use thereof

Country Status (1)

Country Link
CN (1) CN100475835C (en)

Families Citing this family (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008138200A1 (en) * 2007-05-11 2008-11-20 Hong Kong University Of Science & Technology Nmda and mc receptor antagonists exhibiting neuroprotective and memory enhancing activities
CN101066273A (en) * 2007-06-06 2007-11-07 邱日辉 Antitumor medicine containing betulic acid derivative
JP2011020922A (en) * 2007-11-06 2011-02-03 Nihon Univ Agent for treatment of malignant tumor
US9029414B2 (en) * 2008-09-19 2015-05-12 Biotechnology Research Corporation Triterpenoid compounds and methods of use thereof
US9067966B2 (en) 2009-07-14 2015-06-30 Hetero Research Foundation, Hetero Drugs Ltd. Lupeol-type triterpene derivatives as antivirals
CN101880309A (en) * 2010-07-13 2010-11-10 中国药科大学 Semisynthesis method of natural product of 23-hydroxy betulinic acid
CN101880310B (en) * 2010-07-13 2014-05-07 中国药科大学 Split heterocyclic 23-hydroxybetulinic acid derivatives and preparation method, preparation and application thereof
CN101928323B (en) * 2010-07-13 2014-05-07 中国药科大学 23-hydroxy betulinic acid derivative with modified sites 3, 23 and 28 as well as preparation method, preparation and application thereof
CN102108092B (en) * 2011-01-29 2014-07-02 暨南大学 23-hydroxy betulinic acid derivative as well as preparation method and application thereof
WO2014105926A1 (en) 2012-12-31 2014-07-03 Hetero Research Foundation Novel betulinic acid proline derivatives as hiv inhibitors
US20170129916A1 (en) 2014-06-26 2017-05-11 Hetero Research Foundation Novel betulinic proline imidazole derivatives as hiv inhibitors
MA40886B1 (en) 2015-02-09 2020-03-31 Hetero Research Foundation Novel c-3 triterpenone with c-28 reverse amide derivatives as hiv inhibitors
US10370405B2 (en) 2015-03-16 2019-08-06 Hetero Labs Limited C-3 novel triterpenone with C-28 amide derivatives as HIV inhibitors
CN105541955B (en) * 2015-12-16 2017-06-23 中国药科大学 23 hydroxyl radical white birck acid fluorescence probes, preparation and its purposes in cellular localization with intake
CN107501380B (en) * 2016-11-16 2020-09-29 中国药科大学 23-hydroxy betulinic acid 23-position modified derivative, preparation method and application thereof
CN108727460B (en) * 2018-05-19 2019-06-07 河南省科学院高新技术研究中心 Betulonic acid derivative and its synthetic method and application
CN109553591B (en) * 2019-01-15 2020-12-15 山东安信制药有限公司 Preparation method of quetiapine fumarate intermediate

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Medicinal Chemistry Letters,Vol.9 No.8. 1999 *
Preparation of amino acid conjugates of betulinic acid withactivity against human melanoma.. Jeong, Hyeh-Jean, Chai, Hee-Byung, Park, So-Young, Kim,Darrick S. H. L.Bioorganic & Medicinal Chemistry Letters,Vol.9 No.8. 1999 *
Preparation of amino acid conjugates of betulinic acid withactivity against human melanoma.. Jeong, Hyeh-Jean, Chai, Hee-Byung, Park, So-Young, Kim,Darrick S. H. L.Bioorganic &amp *

Also Published As

Publication number Publication date
CN1861627A (en) 2006-11-15

Similar Documents

Publication Publication Date Title
CN100475835C (en) 23-Hydroxyl betulinic acid kind derivant and preparing process, preparation and use thereof
JP6043361B2 (en) Oleanolic acid amidated derivatives, their preparation and use
CN105153122B (en) [(indol-3-yl) pyrimidine -2-base] aminophenyl propyl- 2- alkenylamide derivatives and salt, preparation method, application
CN101230082B (en) 23-hydroxy betulinic acid derivative, preparation method, preparation and use thereof
WO2013044811A1 (en) Gemcitabine amide derivative and preparation method and use thereof
Lan et al. Synthesis and antiproliferative evaluation of 23-hydroxybetulinic acid derivatives
CN108524482A (en) The purposes of 2- (substitution phenylamino) benzoic acids FTO inhibitor for treating leukaemia
JP6211527B2 (en) 2-substituted oleanolic acid derivatives, their preparation and use
CN102796124B (en) Double β carbolines alkaloid compounds, its preparation method and its pharmaceutical composition and purposes
CN105503894A (en) 1-oxo/acylated-14-acylated Oridonin derivative and preparation method and application thereof
CN103694238A (en) NO donor type matrine derivative and preparation method and medical application thereof
CN101157715B (en) Ursolic acid chemical modified compound amino alcohol having antitumor activity
CN104812761B (en) Double B-carboline alkaloid compounds, its preparation method and its pharmaceutical composition and purposes
WO2021000770A1 (en) Heterocyclic compound capable of enhancing immune activity, preparation method therefor and application in medicine
Li et al. Design, synthesis and biological activities of tetrandrine and fangchinoline derivatives as antitumer agents
CN107827875A (en) A kind of application of benzimidazoles derivative as the inhibitor of Cyclin dependent kinase 4/6
CN107556361A (en) Driffractive ring lupinane derivative and its anticancer usage
CN114380864A (en) Dihydroartemisinin derivative, preparation method, pharmaceutical composition and application of dihydroartemisinin derivative in preparation of antitumor drugs
JP2015523349A (en) Acylated derivatives of porphyrin I, methods for their preparation and use
CN101928323B (en) 23-hydroxy betulinic acid derivative with modified sites 3, 23 and 28 as well as preparation method, preparation and application thereof
CN104530171A (en) Pentacyclic triterpenoid compounds, preparation method thereof, and application of compounds to prepare medicines controlling tumors
CN108137644B (en) Compound with anti-tumor effect and preparation method and application thereof
WO2021098716A1 (en) Crystal forms of fused ring compound, and composition thereof, preparation method therefor and application thereof
WO2014154179A1 (en) 2β,3α,5α-TRIHYDROXY-ANDROST-6-ONE AND PREPARATION METHODS AND USE THEREOF
CN102786458B (en) Pyrrole formamide derivative, and preparation method and application thereof

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
C17 Cessation of patent right
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20090408

Termination date: 20120512