CN102558270A - 20(S) and 20(R)-dammarane-3beta,12beta,20,25-tetrol derivative and preparation method and application thereof - Google Patents
20(S) and 20(R)-dammarane-3beta,12beta,20,25-tetrol derivative and preparation method and application thereof Download PDFInfo
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Abstract
The invention belongs to the field of medicine, and relates to a 20(S) and 20(R)-dammarane-3beta,12beta,20,25-tetrol derivative with anti-tumor activity and a preparation method and application thereof. The derivative has a structural formula (I) shown in the specifications, wherein R1 and R2 are mono-substituted or bis-substituted or can be substituted simultaneously; and R1 and R2 are acylation substituted amino acid, amino protected amino acid and oligopeptide, amino protected oligopeptide and polypeptide, amino protected polypeptide, or a base pair which forms deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), and a derivative of the base pair. The anti-tumor activity of the derivative and salt thereof is higher than that of 20(S) and 20(R)-protopanaxadiol, the activity of other anti-tumor medicines can be more obviously enhanced, and the toxic and side effects of the medicines are reduced.
Description
Technical field:
The invention belongs to medical technical field; 20 (S) and 20 (R)-dammarane-3 β with anti-tumor activity are provided, 12 β, 20; The 25-tetraalcohol derivative and its production and application, also relate to this compound and reach with this compound as the purposes of composition of active components at anti-tumor aspect.
Background technology:
20 (R)-dammarane-3; 12; 20; 25-tetrol [20 (R)-25-OH-PPD] has stronger antitumour activity; Can suppress human tumor cells (human breast carcinoma, human small cell lung carcinoma, people's cancer of the stomach, human colon carcinoma, human neuroglia cancer, Humanmachine tumour, human cervical carcinoma, people's liver cancer, early children's grain white blood disease, sarcoma S-180, liver ascites, mouse cervical cancer-14 and ehrlich carcinoma etc.) growth and propagation, inducing tumor cell differentiation, apoptosis suppress tumor neogenetic blood vessels and generate, suppress infiltration and transfer, the enhancing body immunizing power of tumour and reduce anti-tumor activities such as toxic and side.Than the ginsenoside verivate Rg that is widely used at present cancer therapy drug at home
3Growth of tumor is suppressed active high 5-15 doubly.25-OH-PPD can also induce the apoptosis and inhibition cell proliferation of lung cancer cell line H838 (p53 wild-type) and H358 (p53 absence type) with the interdependent mode of dosage; And can make cell-cycle arrest in the G1 phase, this compound of different concns all demonstrates the activity stronger than other ginsenosides.In addition, 25-OH-PPD (when 50 μ M and 100 μ M) effectively reduces the survival rate of prostate cancer cell line LNCaP (p53 wild-type) and PC3 (p53 absence type), but does not influence inoblast; This compound can also be induced the apoptosis of these two kinds of cells, suppresses the propagation of cell, and makes cell-cycle arrest in the G1 phase.In the LNCaP and PC3 cell that 25-OH-PPD handled, the expression of cancer suppressor gene p21, p27 and Bax improves, and the expression of proto-oncogene MDM2, Bcl2, E2F1, cdk2, CDK4 and Cyclin D1 all reduces.25-OH-PPD has stronger anticancer bioactive, but has deficiencies such as poorly water-soluble, bioavailability are low, has had a strong impact on application clinically.Pair cell such as amino acid, peptides, gene base and nucleic acid have good affinity, are introduced in the drug molecule, help medicine and arrive in the cell tissue and play a role effectively.This patent to overcome above-mentioned shortcoming, is found new compound entity efficient, low toxicity through its structure is modified.
Summary of the invention:
The present invention provides 20 (S) and 20 (R)-dammarane-3 β with anti-tumor activity, 12 β, 20,25-tetraalcohol derivative and preparation method thereof.
20 (S) of general formula provided by the invention (I) and 20 (R)-dammarane-3 β, 12 β, 20,25-tetraalcohol derivative and salt thereof:
20 (S) and 20 (R)-dammarane-3 β, 12 β, 20, the general formula of 25-tetraalcohol derivative and salt thereof (I)
Wherein, R1, R2 can be single replacement, two replacement, or replace simultaneously; R1, R2 can be the substituted amino acid of acidylate, the amino acid of amido protecting, oligopeptides, polypeptide, or form base pair and the verivate thereof etc. of DNA, RNA.
20 (S) of general formula provided by the invention (I) and 20 (R)-dammarane-3 β, 12 β, 20,25-tetraalcohol derivative and salt thereof, wherein: described salt is sylvite, sodium salt, ammonium salt, magnesium salts, calcium salt or hydrochloride.
20 (S) provided by the invention and 20 (R)-dammarane-3 β, 12 β, 20, in 25-tetraalcohol derivative and the salt thereof:
Described amino acid is 20 kinds of gal4 amino acids, comprises; L-Ala, Xie Ansuan, leucine, Isoleucine, proline(Pro), phenylalanine(Phe), tryptophane, methionine(Met), Serine, Threonine, halfcystine, tyrosine, l-asparagine, Stimulina, Methionin, l-arginine, Histidine, aspartic acid, L-glutamic acid etc.;
Described base pair comprises VITAMIN B4, guanine, cytosine(Cyt), uridylic, thymus pyrimidine and verivate thereof etc.;
Described amino acid whose D type and the L type of being configured as;
Said oligopeptides is 2～20 peptides that amino-acid residue is connected to form through peptide bond;
The peptide that said polypeptide is made up of the amino-acid residue more than 20.
Described protection base is tertbutyloxycarbonyl (Boc-), carbobenzoxy-(Cbz) (CBz-), fluorenylmethyloxycarbonyl (Fmoc-) etc.
Wherein R1, R2 can singly replace, also can twoly replace.
Formula of the present invention (I) compound can prepare as follows:
20 (S) and 20 (R)-dammarane-3 β, 12 β, 20, the 25-tetraalcohol derivative be corresponding amino acid, oligopeptides and polypeptide under katalysis with 20 (S) and 20 (R)-dammarane-3 β, 12 β, 20, the 25-tetrol carries out acylation reaction, deprotection reaction and makes.The solvent of reaction can be THF, N, benzene, toluene, methylene dichloride, DMSO 99.8MIN. etc.; Temperature of reaction can be 0-150 ℃; The available dewatering agent can be NSC 57182 (DCC), 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (EDC) etc.; The available catalyzer can be 4-Dimethylamino pyridine (DMAP), I-hydroxybenzotriazole (HOBT), triethylamine etc., and available deprotection method has trifluoroacetic acid (TFA), silica gel catalyst, solution method etc.
Synthetic route I:
Each amino acid (b) can be the corresponding amino acid of L-Ala, Xie Ansuan, leucine, Isoleucine, proline(Pro), phenylalanine(Phe), tryptophane, methionine(Met), Serine, Threonine, halfcystine, tyrosine, l-asparagine, Stimulina, Methionin, l-arginine, Histidine, aspartic acid, L-glutamic acid and amido protecting thereof, verivates such as oligopeptides, polypeptide.
Synthetic route II:
Described base pair (M1) comprises VITAMIN B4, guanine, cytosine(Cyt), uridylic, thymus pyrimidine and verivate thereof, like Fluracil, uracil mustard etc.
20 (S) and 20 (R)-dammarane-3 β, 12 β, 20, the salt of 25-tetrol di-carboxylic acid and mineral acid verivate is to transfer pH with the aqueous solution of corresponding oxyhydroxide, gets with organic solvent deposit then.For example use sodium hydroxide, ammoniacal liquor transfers pH to neutral, uses acetone precipitation then, gets corresponding salt.
General structure of the present invention (I) is through to 20 (S) and 20 (R)-dammarane-3 β; 12 β, 20, the 25-tetrol is modified; Introduce some new groups; Particularly introduce and contain water-soluble polar group such as the substituted amino acid derivative of amino-contained,, and strengthen the anti-tumor activity of this compounds the solubleness and the bioavailability that improve verivate.
The invention provides the application of general structure (I) compound in the preparation anti-tumor medicinal preparation.
The invention provides and contain above-mentioned 20 (S) and 20 (R)-dammarane-3 β, 12 β, 20, the medicine of 25-tetraalcohol derivative and salt thereof mixes said derivative and salt thereof and processes preparation with conventional medicinal adjuvant.Said preparation can be granule, capsule, tablet, injection, transfusion or suppository.Medicine of the present invention can be used for treating tumour.
The described compound of general structure (I) can be used for treating various growth of tumor and transfer, and tumour mainly includes: lung cancer, nonsmall-cell lung cancer, liver cancer, carcinoma of the pancreas, cancer of the stomach, osteocarcinoma, esophagus cancer, mastocarcinoma, prostate cancer, carcinoma of testis, colorectal carcinoma, ovarian cancer, bladder cancer, cervical cancer, malignant melanoma, squamous cell carcinoma, rodent cancer, gland cancer, syringocarcinoma, sebaceous carcinoma, papillary carcinoma, papillary carcinoma, cryptomere gland cancer, cystocarcinoma, soft cancer, bronchogenic carcinoma, bone cell cancer, epithelial cancer, cholangiocarcinoma, choriocarcinoma, embryo cancer, spermatogonium cancer, this cancer of Weir nurse, spongiocyte cancer, stellate cell cancer, medulloblast cancer, craniopharyhgeal canal cancer, ependymoma, pinealoma, hemocytoblastoma, vocal cords neuroma, meningioma, neuroblastoma, one-tenth optic cell knurl, neurofibroma, fibrosarcoma, fibroblastoma, fibroma, fibroadenoma, inochondroma, inocystoma, fibromyxoma, fibro-osteoma, fibromyxosarcoma, fibropapilloma, myxosarcoma, myxocystoma, myxochondroma, myxochondrosarcoma, myxochondrofibrosarcoma, myxadenoma, myxoblastoma, liposarcoma, lipoma, lipoadenoma, lipoblastoma, lipochondroma, lipofibroma, lipoangioma, lipomyxoma, chondrosarcoma, chondroma, chondromyoma, chordoma, chorioadenoma, chorioepithelium knurl, chorioblastoma, osteosarcoma, osteoblastoma, osteochondrofibroma, osteochondrosarcoma, osteochondroma, osteocystoma, osteodentinoma, osteofibroma, burst fibrosarcoma, angiosarcoma, vascular tumor, angiolipoma, angiochondroma, hemangioblastoma, angiokeratoma, angioglioma, hemangioendothelioma, hemangiofibroma, angiomyoma, angiolipoma, hematolymphangioma, angiolipoleiomyoma, angiomyoliopma, angiomyoneuroma, angiomyxoma, angioreticuloendothelioma, lymphangiosarcoma, lymph granuloma, lymphangioma, lymphoma, lymphomyxoma, lymphosarcoma, lymphangiofibroma, lymphocytoma, lymphepithelioma, lymphoblastoma, endothelioma, endothelioblastoma, synovioma, synovial sarcoma, mesothelioma, mesocytoma, ewing's tumor, leiomyoma, leiomyosarcoma, leiomyoblastoma, leiomyofibroma, rhabdomyoma, rhabdosarcoma, rhabdomyomyxoma, acute lymphoblastic white blood disease, acute myelogenous leukemia, chronic disease cell, polycyth(a)emia, lymphoma, multiple myeloma.
Dammarane-3 β, 12 β, 20 contained provided by the invention; 25-tetraalcohol derivative and salt thereof are not only than dammarane-3 β, 12 β, 20; The 25-tetrol has stronger antiviral, antitumour activity, can strengthen more significantly also simultaneously that other is antiviral, cancer therapy drug active and reduce these drug toxicities.
Major advantage of the present invention is, at natural product 20 (S) and 20 (R)-dammarane-3 β, and 12 β, 20,3 of the 25-tetrol or 12 introducings contain amino acid, oligopeptides, polypeptide and various base group, and synthesis of derivatives strengthens anti-tumor activity.
At natural product 20 (S) and 20 (R)-dammarane-3 β, 12 β, 20,3 or 12 introducings of 25-tetrol contain the pharmaceutically acceptable salt that forms with alkali or acid neutralization, and the synthetic verivate has improved water-soluble.
Description of drawings:
Fig. 1 is 3 β-N-tertbutyloxycarbonyl-L-alanyl-dammarane-12 β, 20, and 25-triol proton nmr spectra;
Fig. 2 is 3 β-N-tertbutyloxycarbonyl-L-alanyl-dammarane-12 β, 20, and 25-triol carbon-13 nmr spectra;
Fig. 3 is 3 β-L-alanyl-dammarane-12 β, 20, and 25-triol proton nmr spectra;
Fig. 4 is 3 β-L-alanyl-dammarane-12 β, 20, and 25-triol carbon-13 nmr spectra;
Fig. 5 is 3 β-N-tertbutyloxycarbonyl-nitro-L-arginyl-dammarane-12 β, 20, and 25-triol proton nmr spectra;
Fig. 6 is 3 β-N-tertbutyloxycarbonyl-nitro-L-arginyl-dammarane-12 β, 20, and 25-triol carbon-13 nmr spectra;
Fig. 7 is 3 β-L-glycyl-dammarane-12 β, 20, and 25-triol proton nmr spectra;
Fig. 8 is 3 β-L-glycyl-dammarane-12 β, 20, and 25-triol carbon-13 nmr spectra;
Fig. 9 is compound 1-3, the 25-OH-PPD inhibiting rate to MCF-7 clone;
Figure 10 is compound 1-3, the 25-OH-PPD inhibiting rate to A549 clone;
Figure 11 is compound 1-3, the 25-OH-PPD inhibiting rate to Lovo clone.
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in the restriction scope of the present invention.The TP of unreceipted actual conditions in the following example is usually according to the normal condition or the condition of advising according to manufacturer.
Prepare 3 β-N-tertbutyloxycarbonyl-L-alanyl-dammarane-12 β, 20, the 25-triol
Take by weighing 0.3g dammarane-3 β, 12 β, 20, the 25-tetrol is dissolved in the anhydrous CH of 15ml
2Cl
2In, add DCC 0.87g, BOC-L-glycocoll DMAP 0.03g, Boc-L-L-Ala 0.24g, room temperature reaction 24h.Reaction is finished, and filters, and gets filtrate decompression recovery reagent and gets gross product.In the gross product solution 30mL ETHYLE ACETATE, 5%NaHCO
3Extraction (20mL * 3), organic layer is used anhydrous MgSO
4Drying is crossed leaching filtrating, and reclaim under reduced pressure reagent gets the bullion product.Through silica gel column chromatography, use sherwood oil: ETHYLE ACETATE (1:1) wash-out, obtain three components, wherein component 3 is 3 β-N-tertbutyloxycarbonyl-L-alanyl-dammarane-12 β, 20,25-triol (51.5%).
3 β-N-tertbutyloxycarbonyl-L-alanyl-dammarane-12 β, 20,25-triol nuclear magnetic data (accompanying drawing 1,2):
1H-NMR?(75Hz,?C
5D
5N):?δ4.53(1H,H-3),?δ3.58(1H,td,H-12),?δ2.04(3H,s;CH
3),?δ1.44(3H,s;CH
3),?δ1.41(6H,s,?2×CH
3),?δ1.11(3H,s,CH
3),?δ0.99(3H,s,CH
3),?δ0.91(3H,s;CH
3),?δ0.88(3H,?s;CH
3),?δ0.85(3H,s;CH
3);?
13C-NMR?(CDCl
3,?300MHz):?δ?81.8?(C-3),?71.0?(C-12),74.1?(C-20),?70.6?(C-25)，51.5(C-17)，?155.0(N-?C=O)，173.0(-CO-)。
Prepare 3 β-L-alanyl-dammarane-12 β, 20, the 25-triol
Take by weighing 3 β-N-tertbutyloxycarbonyl-L-alanyl-dammarane-12 β, 20,25-triol 0.1g adds in the 25mL round-bottomed flask; Add the dissolving of 10mL toluene, add 0.1g silica gel, reflux 6 hours; Reaction finishes postcooling to room temperature, filters, and uses washed with methanol silica gel; To filtrate behind the evaporate to dryness, get crude product.Through silica gel column chromatography, use methylene dichloride: methyl alcohol (25:1) wash-out obtains end product: buff powder 3 β-L-alanyl-dammarane-12 β, 20,25-triol (57.3%).
3 β-L-alanyl-dammarane-12 β, 20,25-triol nuclear magnetic data (accompanying drawing 3,4):
1H-NMR?(75Hz,C
5D
5N):δ5.75(N-H)，δ4.43(1H,H-3),?δ3.52(1H,td,H-12),?δ2.09(3H,s;?CH
3),?δ1.44(3H,s;CH
3),?δ1.03(3H,s,CH
3),?δ0.93(3H,s,CH
3),?δ0.89(3H,s;CH
3),?δ0.88(3H,s;CH
3),?δ0.83(3H,s;CH
3);?
13C-NMR?(CDCl
3,?300MHz):?δ?80.2?(C-3),?69.7(C-12),?72.3(C-20),?69.0(C-25),?51.1(C-17),?22.9(C-21),?44.7(C-22),?38.8?(C-1),?28.1(C-2),?38.1(C-4),?55.1(C-5),?18.5(C-6),?35.1(C-7),?40.2(C-8),?49.7(C-9),?37.7(C-10),?32.2(C-11),?49.5(C-13),?51.8(C-14),?31.2(C-15),?26.7(C-16),?16.3(C-18),?16.8(C-19),?18.7?(C-23),?45.6(C-24),?30.0(C-26),?30.2(C-27),?14.1(C-28),?15.9?(C-29),?17.4(C-30),?175.4(-CO-)。
Prepare 3 β-N-tertbutyloxycarbonyl-nitro-L-arginyl-dammarane-12 β, 20, the 25-triol
Take by weighing 0.3g dammarane-3 β, 12 β, 20, the 25-tetrol is dissolved in the anhydrous CH of 15ml
2Cl
2, add DCC 0.87g, DMAP 0.03g, Boc-NO
2-L-l-arginine 0.40g, room temperature reaction 24h.Reaction is finished, and filters, and gets filtrate decompression recovery reagent and gets gross product.In the gross product solution 30mL ETHYLE ACETATE, 5%NaHCO
3Extraction (20mL * 3), organic layer is used anhydrous MgSO
4Drying is crossed leaching filtrating, and reclaim under reduced pressure reagent gets the bullion product.Through silica gel column chromatography, use sherwood oil: acetone (1:1) wash-out obtains three components: component 3 is 3 β-N-tertbutyloxycarbonyl-nitro-L-arginyl-dammarane-12 β, 20, and 25-triol (63.5%).
3 β-N-tertbutyloxycarbonyl-nitro-L-arginyl-dammarane-12 β, 20,25-triol nuclear magnetic data (accompanying drawing 5,6):
1H-NMR?(75Hz,?C
5D
5N):?δ4.53(1H,H-3),?δ3.58(1H,td,;H-12),?δ4.28(1H,CH-CO);?
13C-NMR?(CDCl
3,?300MHz):?δ?82.6?(C-3),?71.0?(C-12),74.2?(C-20),?70.6?(C-25)，?60.3(N-C-CO)，155.0(N-?C=O)，172.1(-CO-)。
Prepare 3 β-N-tertbutyloxycarbonyl-D-glycyl-dammarane-12 β, 20, the 25-triol
Take by weighing 0.3g dammarane-3 β, 12 β, 20, the 25-tetrol is dissolved in the anhydrous CH2Cl2 of 15ml, adds DCC 0.87g, DMAP 0.03g, Boc-L-glycocoll 0.22g, room temperature reaction 24h.Reaction is finished, and filters, and gets filtrate decompression recovery reagent and gets gross product.In the gross product solution 30mL ETHYLE ACETATE, 5%NaHCO3 extracts (20mL * 3), and organic layer is dry with anhydrous MgSO4, crosses leaching filtrating, and reclaim under reduced pressure reagent gets the bullion product.Through silica gel column chromatography, use sherwood oil: ETHYLE ACETATE (1:1) wash-out, obtain three components, wherein component 3 is 3 β-N-tertbutyloxycarbonyl-L-glycyl-dammarane-12 β, 20,25-triol (46.5%).
3 β-N-tertbutyloxycarbonyl-L-glycyl-dammarane-12 β, 20,25-triol nuclear magnetic data:
1H-NMR?(75Hz,?C
5D
5N):?δ4.56(1H,H-3),?δ3.61(1H,td;H-12),?δ2.05(3H,s;CH
3),?δ1.44(3H,s;CH
3),?δ1.43(6H,s,?2×CH
3),?δ1.03(3H,s,CH
3);
13C-NMR?(CDCl
3,?300MHz):?δ?81.8?(C-3),?71.0?(C-12),74.1?(C-20),?70.6?(C-25),?155.1(N-?C=O)，172.9(-CO-)。
Prepare 3 β-D-glycyl-dammarane-12 β, 20, the 25-triol
Take by weighing 3 β-N-tertbutyloxycarbonyl-D-glycyl-dammarane-12 β, 20,25-triol 0.1g adds in the 25mL round-bottomed flask; Add the dissolving of 10mL toluene, add 0.1g silica gel, reflux 6 hours; Reaction finishes postcooling to room temperature, filters, and uses washed with methanol silica gel; To filtrate behind the evaporate to dryness, get crude product.Through silica gel column chromatography, use methylene dichloride: methyl alcohol (25:1) wash-out obtains end product: buff powder 3 β-L-alanyl-dammarane-12 β, 20,25-triol (52.6.3%).
3 β-D-glycyl-dammarane-12 β, 20,25-triol nuclear magnetic resonance spectrum data:
1H-NMR?(75Hz,?C
5D
5N)?:δ5.75(N-H),?δ4.45(1H,H-3),?δ3.52(1H,H-12),?δ2.08(3H,s;?CH
3),δ0.88(3H,?s;CH
3),?δ0.83(3H,s;CH
3);?
13C-NMR?(CDCl
3,?300MHz):?δ?80.2?(C-3),?69.7(C-12),72.3(C-20),?69.0(C-25);?51.5(C-17),?22.9(C-21),?44.1(C-22),?38.6?(C-1),?28.1(C-2),?38.1(C-4),?55.1(C-5),?18.5(C-6),?35.1(C-7),?40.1(C-8),?49.9(C-9),?37.2(C-10),?32.2(C-11),?49.5(C-13),?51.8(C-14),?31.5(C-15),?26.7(C-16),?16.3(C-18),?16.8(C-19),?18.7?(C-23),?45.6(C-24),?30.0(C-26),?30.2(C-27),?14.1(C-28),?15.9?(C-29),?17.4(C-30),?173.9(-CO-)。
Prepare 3 β-N-tertbutyloxycarbonyl-L-Xie Ansuan base-dammarane-12 β, 20,25-triol, 12 β-N-tertbutyloxycarbonyl-L-be valyl-dammarane-3 β, 20, the 25-triol
Take by weighing 0.3g dammarane-3 β, 12 β, 20, the 25-tetrol is dissolved in the anhydrous CH of 15ml
2Cl
2, add DCC 0.87g, BOC-L-glycocoll DMAP 0.03g, Boc-L-Xie Ansuan 0.27g, room temperature reaction 24h.Reaction is finished, and filters, and gets filtrate decompression recovery reagent and gets gross product.In the gross product solution 30mL ETHYLE ACETATE, 5%NaHCO
3Extraction (20mL * 3), organic layer is used anhydrous MgSO
4Drying is crossed leaching filtrating, and reclaim under reduced pressure reagent gets the bullion product.Through silica gel column chromatography, use sherwood oil: ETHYLE ACETATE (1:1) wash-out obtains three components; Wherein component 2 is white solid 12 β-N-tertbutyloxycarbonyl-L-valyl-dammarane-3 β; 20,25-triol (22.8%), component 3 be 3 β-N-tertbutyloxycarbonyl-L-valyl-dammarane-12 β; 20,25-triol (58.5%).
3 β-N-tertbutyloxycarbonyl-L-is valyl-dammarane-12 β, and 20,25-triol nuclear magnetic data:
1H-NMR?(75Hz,?C
5D
5N):?δ5.03(N-H),?δ4.56(1H,H-3),?δ3.59(1H,;H-12),?δ0.89(3H,s;CH
3),?δ0.88(3H,?s;CH
3),?δ0.83(3H,s;CH
3);?
13C-NMR?(CDCl
3,?300MHz):?δ?82.1?(C-3),?71.0?(C-12),74.2?(C-20),?70.7?(C-25),?79.6(C-O),?155.8(N-?C=O),?172.7(-CO-)。
12 β-N-tertbutyloxycarbonyl-L-is valyl-dammarane-12 β, and 20,25-triol nuclear magnetic data:
1H-NMR?(75Hz,?C
5D
5N):?δ5.34(N-H),?δ4.56(1H,H-3),?δ3.47(1H,;H-12),?δ0.89(3H,s;CH
3),?δ0.88(3H,?s;CH
3),?δ0.83(3H,s;CH
3);?
13C-NMR?(CDCl
3,?300MHz):?δ?78.7?(C-3),?76.6?(C-12),73.7?(C-20),?71.1?(C-25),?79.6(C-O),?155.8(N-?C=O),?172.5(-CO-)。
Preparation: 5 FU 5 fluorouracil-1-guanidine-acetic acid is synthetic
(5.20g 0.04mol) is dissolved in the NaOH solution of 40mL0.5mol/L with 5 FU 5 fluorouracil.Controlled temperature 40-50 ℃, (5.64g, 0.06mol) solution 30mL dripped off in 40 minutes to stir slow down dropping Mono Chloro Acetic Acid.Control under this temperature and react, thin-layer chromatography is followed the tracks of reaction, and the 5-FU raw material reaction was complete in about 6 hours.Mixed solution is cooled to room temperature, regulates pH to 1-2, hide in refrigerator and cooled and spend the night with hydrochloric acid.Filter, throw out is used cold water washing, dry compound 5 FU 5 fluorouracil-1-guanidine-acetic acid (5-FUAA), yield 82%.
Preparation: 20 (R)-3 β-5
'-Fluracil-1 '-Ji-ethanoyl-dammarane-12 β, 20,25-triol synthetic
Take by weighing 25-OH-PPD 0.9g (1.883mmol) and add among the 50mLDMF, add BOC-L-L-Ala 071g (3.766mmol) then, jolting makes dissolving; In ice-water bath, add DCC 1.164g (3.766mmol), DMAP 0.5g (4.093mmol), room temperature reaction 24h filters; Filtrate decompression reclaims solvent to doing; (2.0 * 50cm) chromatographies with ETHYLE ACETATE-methyl alcohol (50:1 wash-out), separate obtaining compound 4 (51%) to silicagel column on the dry method.
20 (R)-3 β-5 '-Fluracil-1 '-Ji-ethanoyl-dammarane-12 β, 20,25-triol nuclear magnetic data (accompanying drawing 7,8):
1H-NMR (75Hz, C
5D
5N):
δ10.02 (1H, s),
δ7.3 (1H, s), δ 4.58 (1H, H-3), δ 3.59 (1H; H-12), δ 0.89 (3H, s; CH
3), δ 0.88 (3H, s; CH
3), δ 0.83 (3H, s; CH
3);
13C-NMR (CDCl
3, 300MHz): δ 83.6 (C-3), 71.1 (C-12), 74.3 (C-20), 70.5 (C-25),
δ166.8 (C=O),
δ149.6 (C=O),
δ157.5 (C=O).
Active testing
Dammarane-3 β, 12 β, 20, the test of 25-tetraalcohol derivative anti-tumor activity
Adopt classical mtt assay to carry out the anti-tumor activity test, to we synthetic dammarane-3 β, 12 β; 20,25-tetraalcohol derivative and control compound Fluracil (5-Fu) are like human breast cancer cell MCF-7; The human lung cancer cell A549; Human colon cancer cell Lovo, most verivates such as human cervical carcinoma cell strain HeLa, large bowel cancer HCT116, cerebral glioma U87, human large intestine cancer HT29 have the broad-spectrum anti-tumor effect, and have confirmed IC
50Value.
Main agents and major equipment: RMPI RPMI-1640 (available from Gibco); The high sugared nutrient solution (available from Gibco) of DMEM; DMEM low sugar nutrient solution (available from Gibco); Calf serum (available from Hyclone); Foetal calf serum (TBD company); Tetramethyl-azo azoles salt (MTT) U.S. Sigma (St. Louis, MO); Trypsin Biosharp); DMSO (Shenyang chemical reagent factory); NaCl (Shenyang chemical reagent factory); KCl (Shenyang chemical reagent factory); KH2PO3 (Shenyang chemical reagent factory); Na2HPO3 (Shenyang chemical reagent factory); NaHCO3 (Shenyang chemical reagent factory); ELIASA (Austrian TECAN); 96 porocyte culture plates (Costar company).
Experimental technique: the culture condition of human colon cancer cell Lovo cell strain, human cervical carcinoma cell strain HeLa cell strain, human lung cancer cell A549's cell strain is the RMPI RPMI-1640; 10% foetal calf serum, the culture condition of human breast cancer cell MCF-7 cell strain, large bowel cancer HCT116 cell strain, cerebral glioma U87 cell strain, human large intestine cancer HT29 cell strain are low sugar DMEM nutrient solution 10% foetal calf serum.Above cell strain all is incubated at 5%CO
2, in 37 ℃ the incubator.Use the DMSO dissolving, be made into the mother liquor that concentration is 10mmol/L, be stored in-20 ℃.
The RMPI RPMI-1640 of cell strain use serum-free is diluted to 1000umol/L with the mother liquor of medicine, 300umol/L, and 100umol/L, 10umol/L, every hole adds 10uL, makes that the final concentration of medicine is 100umol/L, 30umol/L, 10umol/L, 1umol/L.
As shown in Figure 9,3 β-L-alanyl-dammarane-12 β, 20,25-triol (compound 1) acts on the dose response curve of breast cancer cell line MCF-7, cultivates 48 hours IC
50=2.9 μ M are starkly lower than 20 (R)-dammarane-3 β, 12 β, 20,25-tetrol IC
50=48.9 μ M promptly are active 16 times of lead compounds.3 β-N-tertbutyloxycarbonyl-nitro-L-arginyl-dammarane-12 β, 20,25-triol (compound 2), 20 (R)-3 β-5
'-Fluracil-1 '-Ji-ethanoyl-dammarane-12 β, 20,25-triol (compound 3) acts on the IC of MCF-7
50Be respectively 6.8 μ M, 38.1 μ M, all be lower than lead compound 25-OH-PPD.
Equally, compound 1-3 is for 48 hours IC of human lung cancer cell A549's cell strain
50Be respectively 2.8,10.2,38.9 μ M, anti-tumor activity be significantly increased (Figure 10).For 48 hours IC of human colon cancer cell Lovo cell strain
50Be respectively 9.9,0.5,35.2 μ M, anti-tumor activity all is better than 25-OH-PPD (Figure 11).Meanwhile, find that the compound with general formula (I) has the characteristics of selective killing tumour cell, and active 20 (R)-dammarane-3 β, 12 β, 20, the 25-tetrol of being better than more.
FORMULATION EXAMPLE 1
Tablet
Every contains compound 3 β-L-alanyl-dammarane-12 β, and 20, the tablet of 25-triol (preparation embodiment 2) 5mg is by following formulation.With following component uniform mixing, be pressed into the tablet of every heavy 170mg with ordinary method.
FORMULATION EXAMPLE 2
Hard capsule
Every contains compound 3 β-L-alanyl-dammarane-12 β, and 20, the hard capsule of 25-triol (preparation embodiment 2) 5mg is by following prescription, uniform mixing, the hard capsule of packing into.
FORMULATION EXAMPLE 3
Powder
Every contains compound 3 β-L-alanyl-dammarane-12 β, and 20, the powder of 25-triol (preparation embodiment 2) 5mg is by following prescription, uniform mixing, every packed 1.0g powder.
FORMULATION EXAMPLE 4
Injection
Every milliliter of injection liquid contains compound 3 β-L-alanyl-dammarane-12 β, and 20, the injection of 25-three alcohol hydrochlorides (preparation embodiment 2) 5mg is by following prescription, prepares with sterile distilled water.
Above embodiment can more specify the present invention, but does not limit the present invention in any form.
Claims (8)
1. 20 (S) and 20 (R)-dammarane-3 β that have general formula (I), 12 β, 20,25-tetraalcohol derivative and salt thereof:
（I）
Wherein, R1, R2 are single replacement, two replacement, or replace simultaneously; R1, R2 are the substituted amino acid of acidylate, the amino acid of amido protecting, oligopeptides, polypeptide, or form base pair and the verivate thereof of DNA, RNA.
2. 20 (S) of the described general formula of claim 1 (I) and 20 (R)-dammarane-3 β, 12 β, 20,25-tetraalcohol derivative and salt thereof, wherein:
Described salt is sylvite, sodium salt, ammonium salt, magnesium salts, calcium salt or hydrochloride.
3. 20 (S) of the described general formula of claim 1 (I) and 20 (R)-dammarane-3 β, 12 β, 20,25-tetraalcohol derivative and salt thereof, wherein:
Described amino acid is selected from L-Ala, Xie Ansuan, leucine, Isoleucine, proline(Pro), phenylalanine(Phe), tryptophane, methionine(Met), Serine, Threonine, halfcystine, tyrosine, l-asparagine, Stimulina, Methionin, l-arginine, Histidine, aspartic acid, L-glutamic acid.
4. 20 (S) of the described general formula of claim 1 (I) and 20 (R)-dammarane-3 β, 12 β, 20,25-tetraalcohol derivative and salt thereof, wherein:
Said oligopeptides is 2-20 the peptide that amino-acid residue is connected to form through peptide bond;
The peptide that said polypeptide is made up of the amino-acid residue more than 20.
5. 20 (S) of the described general formula of claim 1 (I) and 20 (R)-dammarane-3 β, 12 β, 20,25-tetraalcohol derivative and salt thereof, wherein:
Described amino acid whose D type or the L type of being configured as;
The amino acid whose protection base of described amido protecting is selected from tertbutyloxycarbonyl, carbobenzoxy-(Cbz), fluorenylmethyloxycarbonyl;
Described base pair is selected from VITAMIN B4, guanine, cytosine(Cyt), uridylic, thymus pyrimidine and verivate thereof.
6. 20 (S) of the described general formula of claim 1 (I) and 20 (R)-dammarane-3 β, 12 β, 20,25-tetraalcohol derivative and salt thereof, wherein: R1, R2 are single the replacement or two replacement.
7. compsn comprises any one described 20 (S) of claim 1-6 and 20 (R)-dammarane-3 β of significant quantity, 12 β, 20,25-tetraalcohol derivative and salt thereof and its pharmaceutically acceptable carrier.
8. any one described 20 (S) and 20 (R)-dammarane-3 β among the claim 1-6,12 β, 20, the application of the described compsn of 25-tetraalcohol derivative and salt thereof and claim 7 in preparation treatment antitumor drug.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103724392A (en) * | 2013-12-18 | 2014-04-16 | 江南大学 | Preparation method of amino acid phytosterol ester hydrochloride |
| CN104523702A (en) * | 2014-12-17 | 2015-04-22 | 吉林农业大学 | Application of phenylalanine cyclopentanoperhydro-phenanthrene ester in preparation of hypotensor |
| CN105693809A (en) * | 2016-01-13 | 2016-06-22 | 华中农业大学 | Compound with anti-tumor activity and application of compound |
| CN113527399A (en) * | 2021-06-08 | 2021-10-22 | 陕西巨子生物技术有限公司 | Ginsenoside CK derivative and application thereof in preparation of antitumor drugs |
| CN115894596A (en) * | 2022-12-15 | 2023-04-04 | 湖南医药学院 | Nitrate NO donor type panaxadiol derivative and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20010034363A1 (en) * | 1996-03-12 | 2001-10-25 | Chun Li | Water soluble paclitaxel derivatives |
| CN101250212A (en) * | 2008-04-15 | 2008-08-27 | 沈阳药科大学 | 20(S) or 20(R)-dammarane-3 beta,12 beta, 20,25-tetraalcohol derivative, its salt and use thereof |
| CN102093453A (en) * | 2006-08-07 | 2011-06-15 | 辽宁新中现代医药有限公司 | Method for preparing 20(R)-25-hydroxy-dammarane type-3beta,12beta,20-triols |
-
2012
- 2012-03-09 CN CN2012100610338A patent/CN102558270A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20010034363A1 (en) * | 1996-03-12 | 2001-10-25 | Chun Li | Water soluble paclitaxel derivatives |
| CN102093453A (en) * | 2006-08-07 | 2011-06-15 | 辽宁新中现代医药有限公司 | Method for preparing 20(R)-25-hydroxy-dammarane type-3beta,12beta,20-triols |
| CN101250212A (en) * | 2008-04-15 | 2008-08-27 | 沈阳药科大学 | 20(S) or 20(R)-dammarane-3 beta,12 beta, 20,25-tetraalcohol derivative, its salt and use thereof |
Non-Patent Citations (2)
| Title |
|---|
| 刘继华 等: "20(S)-原人参二醇氨基酸衍生物的合成", 《中国现代应用药学》 * |
| 刘继华: "具有抗癌活性人参皂苷的结构修饰研究", 《吉林大学博士学位论文》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103724392A (en) * | 2013-12-18 | 2014-04-16 | 江南大学 | Preparation method of amino acid phytosterol ester hydrochloride |
| CN103724392B (en) * | 2013-12-18 | 2016-03-02 | 江南大学 | A kind of preparation method of amino acid phytosterol ester hydrochloride |
| CN104523702A (en) * | 2014-12-17 | 2015-04-22 | 吉林农业大学 | Application of phenylalanine cyclopentanoperhydro-phenanthrene ester in preparation of hypotensor |
| CN105693809A (en) * | 2016-01-13 | 2016-06-22 | 华中农业大学 | Compound with anti-tumor activity and application of compound |
| CN113527399A (en) * | 2021-06-08 | 2021-10-22 | 陕西巨子生物技术有限公司 | Ginsenoside CK derivative and application thereof in preparation of antitumor drugs |
| CN115894596A (en) * | 2022-12-15 | 2023-04-04 | 湖南医药学院 | Nitrate NO donor type panaxadiol derivative and preparation method and application thereof |
| CN115894596B (en) * | 2022-12-15 | 2024-02-09 | 湖南医药学院 | Nitric acid ester NO donor type ginseng diol derivative and preparation method and application thereof |
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