CN105622704A - Preparation method and application of antitumor drug X-TOA - Google Patents
Preparation method and application of antitumor drug X-TOA Download PDFInfo
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Abstract
The invention provides a preparation method of compounds with the structure represented by general formula 1, and an application of the compounds in the preparation of antitumor drugs. The compounds are prepared through ester bond condensation of a 3th hydroxyl group of an anticancer lead compound TOA (patent 201110055102.X) and a carboxyl group of amino acids; and the compounds have a more substantial inhibition effect on liver cancer, colorectal carcinoma, cervical carcinoma and stomach cancer cell lines than the TOA, have smaller toxicity to normal cells than cis-platinum, have obviously better water solubility than the TOA, and are especially suitable for preparing antitumor treatment drugs.
Description
Technical field
The present invention relates to pharmaceutical chemistry and area of pharmacology, be specifically related to X-TOA general structure and synthesis thereof and application, pharmacological evaluation proves that this compounds has obvious antitumor action, and normal cytotoxicity is only small; Compare precursor compound TOA and improved in the toxicity of the cell to tumor, meanwhile, improve its hydrophilic. Can be used for the cancer such as preparation prevention and treatment hepatocarcinoma.
X-TOA general structure
Wherein, R represents glycine, alanine, proline, leucine, isoleucine, valine, lysine, phenylalanine, sarcosine, pyroglutamic acid, tryptophan and serine.
Background technology
Chinese patent ZL201110055102.X discloses anticancer lead compound TOA, and pharmacodynamic experiment shows, the in-vitro multiplication of the primer TOA more significant suppression kinds of tumor cells of energy, and normal cytotoxicity is relatively low; The internal growth to S180 murine sarcoma has a remarkable inhibiting activity, and can improve the spleen index of tumor-bearing mice, all in dose-dependence. By calculating tumor cross-sectional area, it is possible to be clearly observable gross tumor volume, tumor cross-sectional area ratio is obviously reduced, and toxic and side effects is substantially less than cyclophosphamide. The research of tumor death NF-�� B signal path is found, the expression of sarcoma mice P65, COX-2 and VEGF can be significantly reduced after TOA administration, and the activity of metalloproteases-2 can be suppressed; Additionally, TOA also can substantially suppress chick chorioallantoic membrane neovascularization resulting, there is the effect of angiogenesis inhibitor. Odd-numbered day gives TOA maximum tolerated dose 6000mg kg continuously-1, in observing 14 days, there is not toxic reaction in mice, is better than cyclophosphamide. But, the shortcoming of TOA poorly water-soluble affects its oral absorption availability. By galenic pharmacy method (such as liposome, microemulsion), TOA is carried out inclusion, its bioavailability be improved significantly; TOA/PVP solid dispersion dissolution rate significantly improves, and its relative bioavailability improves 4 times; The relative bioavailability of TOA microemulsion also improves 57 times. But, improve TOA water solublity by chemical method and improve bioactive research and not yet carry out. Big quantity research shows, aminoacid has hydrophilic, has good effect in improving the water solublity of hydrophobic drug, raising biological activity.
The present invention carries out chemistry split synthesis the compounds of this invention with antitumor lead compound (TOA) and aminoacid for initiation material. The activity rating of this compounds is launched mainly around anti-tumor activity, tests the analog cytotoxic activity to 4 kinds of cancer cell systems (HepG2, HT-29, Hela, BGC-823) and dog renal epithelial cell (MDCK) respectively; The ClogP of compositions is simulated by computer software; It addition, by the method evaluation G-TOA of Giemsa and the DAPI dyeing impact on HePG2 cellular morphology.
Summary of the invention
An object of the present invention is to provide compounds X-TOA general structure (formula 1).
The preparation method that the two of the purpose of the present invention are to provide X-TOA.
The three of the purpose of the present invention are to provide X-TOA application in preparing antitumor drug.
Wherein, R represents glycine, alanine, proline, leucine, isoleucine, valine, lysine, phenylalanine, sarcosine, pyroglutamic acid, tryptophan and serine.
The compounds of this invention numbering and structural formula are as follows:
The compounds of this invention is prepared by following three kinds of methods:
(1) TOA being dissolved in organic solvent, with the condensation under condensing agent, catalysts conditions of Boc-aminoacid, then in acid condition, de-Boc-protects base to generate G-TOA, A-TOA, P-TOA, L-TOA, I-TOA, V-TOA;
(2) TOA is dissolved in organic solvent, with the condensation under condensing agent, catalysts conditions of Cbz-aminoacid, then Pd/C-H2Under condition, de-Cbz-protects base to generate K-TOA, F-TOA, Sa-TOA, Py-TOA, W-TOA;
(3) TOA is dissolved in organic solvent, with TBDMS-Cl protection the condensation under condensing agent, catalysts conditions of Cbz-L-serine, then with TBAF take off TBDMS-protect base, last Pd/C-H2Under condition, de-Cbz-protects base to generate S-TOA;
In above-mentioned preparation method, described reaction carries out in the mixed solvent of dichloromethane, ethyl acetate, methanol or their various ratios; Relating to condensation reaction to carry out at 20-30 DEG C, catalyst is DMAP (DMAP), and condensing agent is 1-ethyl-3-(3-dimethylamine propyl) carbodiimide hydrochloride (EDCI); Boc-protects base deprotection to carry out in cryosel is bathed; Cbz-protects base deprotection to carry out at 20-30 DEG C; The protection of the hydroxyl of Cbz-L-serine is carrying out at 20-30 DEG C, and deprotection carries out in cryosel is bathed; It addition, in acid used, mineral acid is hydrochloric acid.
Further, in above-mentioned preparation method, the mol ratio of corresponding amino acid starting material and TOA is 2��1.5: 1; The mol ratio of condensing agent (EDCI) and TOA is 2��1.5: 1; The mol ratio of catalyst (DMAP) and TOA is 0.5��0.1: 1; Step a in said method, b, c, d, e reaction equation be:
Reaction condition and reagent: a.EDCI, DAMP, 20-30 DEG C of stirring 12h; B.3MHCl/ ethyl acetate, 20-30 DEG C of stirring 1h; Or Pd (OH) c.10%Pd/C2/ C, H2, 20-30 DEG C of stirring 12h; D.TBDMS-Cl, imidazoles, DMF, 20-30 DEG C of stirring 12h; E.TBAF, cryosel is bathed, 0.5h.
The present invention also provides for the application in preparing antitumor drug of formula 1,2 compound.
Further, described tumor is hepatocarcinoma, gastric cancer, colon cancer, cervical cancer;
Present invention also offers a kind of antitumor medicine composition, said composition contained 1 or formula 2 compound and pharmaceutically acceptable carrier. Described pharmaceutical composition can be the dosage forms such as tablet, capsule, granule, powder, oral liquid, injection, liposome, microemulsion.
The compounds of this invention has the activity substantially suppressing tumor cell line (HepG-2, HT-29, Hela, BGC-823) to grow, compared with cisplatin, the toxicity of normal cell (dog renal epithelial cell system MDCK) is less; Wherein, compound G-TOA, A-TOA, P-TOA and K-TOA anti-tumor activity be better than positive drug TOA and cisplatin, and G-TOA, A-TOA and P-TOA nephrotoxicity is weaker than cisplatin.
Experimental example mtt assay observes the present composition X-TOA proliferative effect to cancerous cell and dog renal epithelial cell
1. instrument and material
Thermo3111 type CO2Incubator; HFsafe Biohazard Safety Equipment; MultiskanGO microplate reader; Board LD5-2B type table-type low-speed centrifuge is found in capital; OlympusIX71 inverted fluorescence microscope; Modified form RPMI-1640 culture medium, hyclone, 0.25% trypsin solution, tetrazolium bromide, phosphate buffer (Baeyer enlightening bio tech ltd, Beijing); Amresco dimethyl sulfoxide (DMSO);
Human hepatoma cell line HepG2; CCL188 HT-29; Human cervical cancer cell lines Hela SGC-7901 BCG-823; Dog renal epithelial cell system MDCK;
Experimental agents: the compounds of this invention 1-12 (is prepared by embodiment 2-13) respectively; Reaction raw materials Boc-aminoacid, Cbz-aminoacid (Beijing coupling Science and Technology Ltd., Beijing Yi Nuokai Science and Technology Ltd.); Positive drug cisplatin for injection (301001CF; Qilu Pharmaceutical Co., Ltd.); T-OA (is prepared by 201110055102.X Chinese patent).
2. method
The cultivation of 2.1 different cell strains
HepG2, HT-29, Hela, BCG-823, mdck cell are cultivated in the RPMI-1640 containing 10% hyclone, are positioned over 37 DEG C, the CO of 5%2Incubation in incubator. Cell all grows in adhered state, observes upgrowth situation, Secondary Culture when cell quantity is appropriate under inverted microscope.
2.2 primary dcreening operation cell inhibitory rates
Take the logarithm the HepG2 of trophophase, HT-29, Hela, BCG-823, mdck cell, with 3 �� 103The quantity in/hole is inoculated in 96 well culture plates, containing 5%CO2Humidified incubator in 37 DEG C cultivate 24h. Every hole is separately added into 100 �� L testing compounds, makes every hole concentration respectively 20 ��Ms and 40 ��Ms. Arranging cell controls group and blank group, the every concentration of medicine group repeats 4 holes, cell controls group and blank group and repeats 4 holes. After incubator continues cultivation 72h, every hole adds 20 �� LMTT and hatches 4h, supernatant discarded, add 150 �� LDMSO again, vibration 10min, microplate reader 490nm wavelength records absorbance and records result, it is suppressed that rate/%=[1-(it is blank that A is administered-A)/(A is normal, and-A is blank)] �� 100%. By under 20 ��Ms of concentration, it is suppressed that the rate compound more than 50% carries out multiple sieve, calculate IC50Value.
2.3 multiple sieve cell suppression ratio
Operational approach such as 2.2, the HepG2 of trophophase of taking the logarithm, HT-29, Hela, BCG-823, mdck cell, with 3 �� 103The quantity in/hole is inoculated in 96 well culture plates, containing 5%CO2Humidified incubator in 37 DEG C cultivate 24h; Every hole is separately added into 100 �� L testing compounds, makes ultimate density respectively 20 ��Ms, 10 ��Ms, 5 ��Ms, 2.5 ��Ms, 1.25 ��Ms, 0.625 ��M. Arranging cell controls group and blank group, the every concentration of medicine group repeats 4 holes, cell controls group and blank group and repeats 4 holes. After continuing cultivation 72h in incubator, every hole adds 20 �� LMTT and hatches 4h, supernatant discarded, then adds 150 �� LDMSO, and vibration 10min, microplate reader 490nm wavelength records absorbance, records result the IC of computerized compound50Value, cell inhibitory rate (%)=[1-(it is blank that A is administered-A)/(A is normal, and-A is blank)] �� 100%.
3. result
The compounds of this invention 1-12 and reaction raw materials TOA and the positive drug (cisplatin) IC to 4 kinds of tumor cell lines (HepG2, HT-29, Hela, BCG-823) and normal cell (MDCK)50Value is as shown in table 1.
As can be seen from Table 1, various cancerous cell are all shown the effect of good Inhibit proliferaton by compound G-TOA, A-TOA, P-TOA, K-TOA, Sa-TOA, Py-TOA and S-TOA; And compared with TOA, K-TOA shows higher anti tumor activity in vitro; Compared with cisplatin, G-TOA, A-TOA and P-TOA are less to Normocellular toxicity. Wherein, G-TOA, A-TOA, P-TOA and K-TOA are inhibited to the propagation of hepatoma carcinoma cell.
Table 1 different pharmaceutical IC to different tumor cell lines50Value
Indicate: "-a" show IC50> 25.0 ��Ms, namely that the inhibitory activity of tumor cell is more weak, it does not have to study further.
Experimental example computer calculates the ClogP of the compounds of this invention
The ClogP being calculated the compounds of this invention by computer is as shown in table 1. G-TOA, A-TOA, K-TOA, Py-TOA and S-TOA show the ClogP value lower than TOA. Experiment proves: little molecule aminoacid, polarity or basic amino acid all can reduce the ClogP value of TOA. Therefore, the compound of hydrophilic difference can improve its water solublity and cytotoxic activity by amino acid whose introducing, and this is consistent with literature research report.
Experimental example Giemsa staining observes the compounds of this invention G-TOA impact on HepG2 and mdck cell form
1. instrument and material
Thermo3111 type CO2Incubator; HFsafe Biohazard Safety Equipment; MultiskanGO microplate reader; OlympusIX71 inverted fluorescence microscope; Modified form RPMI-1640 culture medium, hyclone, Jim Sa dyestuff, phosphate buffer (PBS) (Baeyer enlightening bio tech ltd, Beijing); Ethanol; Water;
Human hepatoma cell line HepG2; Dog renal epithelial cell system MDCK;
Experimental agents: G-TOA (is prepared by embodiment 2); Positive drug cisplatin for injection (CDDP, 301001CF; Qilu Pharmaceutical Co., Ltd.);
2. method
The cultivation of 2.1 different cell strains
HepG2, mdck cell are cultivated in the RPMI-1640 containing 10% hyclone, are positioned over 37 DEG C, the CO of 5%2Incubation in incubator. Cell all grows in adhered state, observes upgrowth situation, Secondary Culture when cell quantity is appropriate under inverted microscope.
2.2Giemsa dyes
2.2.1HepG2 cellular morphology is observed
Take the logarithm the HepG2 cell of trophophase, with 3 �� 103The quantity in/hole is inoculated in 12 well culture plates, containing 5%CO2Humidified incubator in 37 DEG C cultivate 24h. Every hole is separately added into 150 �� LG-TOA, makes ultimate density respectively 0 ��M, 1 ��M, 2 ��Ms, 4 ��Ms, and every concentration repeats 3 holes. Incubator continues cultivate after 72h, discard cell culture fluid, with PBS twice, with PBS: ethanol (1: 1) infiltration 2min, discard, with the fixing 10min of ethanol, discard, Giemsa dyeing 2min, clear water washes away dyestuff, in the lower observation of cell form of inverted microscope 400 times;
2.2.2MDCK cellular morphology is observed
Take the logarithm the mdck cell of trophophase, with 3 �� 103The quantity in/hole is inoculated in 12 well culture plates, containing 5%CO2Humidified incubator in 37 DEG C cultivate 24h. Add 2 ��Ms of G-TOA150 �� L, cell controls group and positive group (CDDP), medicine group, cell controls group and each repetition 3 hole of positive group are set. Incubator continues cultivate after 72h, discard cell culture fluid, with PBS twice, with PBS: ethanol (1: 1) infiltration 2min, discard, with the fixing 10min of ethanol, discard, Giemsa dyeing 2min, clear water washes away dyestuff, in the lower observation of cell form of inverted microscope 200 times;
3. result
3.1HepG2 cellular morphology is observed
The change of G-TOA inducing cell form as shown in Figure 1, the change of G-TOA inducing cell form, painted shoal including the forfeiture of iuntercellular contact, core, karyolysis, the increasing of cell debris; Along with the increase of administration concentration, cellular morphology damage increases the weight of; When administration concentration is 2 ��Ms and 4 ��Ms, it does not have the cell of intact form exists.
3.2MDCK cellular morphology is observed
As shown in Figure 2, normal group cell volume is less, and cell is intensive in the change of G-TOA and CDDP inducing cell form; Administration group (G-TOA) cell density is compared normal group and is diminished, but can keep normal cellular morphology, and positive group (CDDP) cell volume increases, and nucleus is painted to shoal, and the cell opposite sex occurs; Therefore, compared with CDDP, the nephrotoxicity of G-TOA is less.
Experimental example DAPI staining observes the compounds of this invention G-TOA impact on HepG2 karyomorphism
1. instrument and material
Thermo3111 type CO2Incubator; HFsafe Biohazard Safety Equipment; MultiskanGO microplate reader; OlympusIX71 inverted fluorescence microscope; Modified form RPMI-1640 culture medium, hyclone, DAPI dyestuff, phosphate buffer (PBS) (Baeyer enlightening bio tech ltd, Beijing); Ethanol; Water;
Human hepatoma cell line HepG2;
Experimental agents: G-TOA (is prepared by embodiment 2) respectively;
2. method
The cultivation of 2.1 different cell strains
HepG2 cell is cultivated in the RPMI-1640 containing 10% hyclone, is positioned over 37 DEG C, the CO of 5%2Incubation in incubator. Cell all grows in adhered state, observes upgrowth situation, Secondary Culture when cell quantity is appropriate under inverted microscope.
2.2DAPI dyes
2.2.1HepG2 karyomorphism is observed
Take the logarithm the HepG2 cell of trophophase, with 3 �� 103The quantity in/hole is inoculated in 12 well culture plates, containing 5%CO2Humidified incubator in 37 DEG C cultivate 24h. Every hole is separately added into 150 �� L testing compound (G-TOA), makes ultimate density respectively 0 ��M, 1 ��M, 2 ��Ms, 4 ��Ms, and every concentration repeats 3 holes. Incubator continues cultivate after 72h, discard cell culture fluid, with PBS twice, with the fixing 15min of 4% paraformaldehyde (pH7.4), discard fixative, with PBS twice, DAPI dyeing 2min, in the lower observation of cell nuclear morphology of fluorescence microscope 400 times;
3. result
3.1HepG2 cellular morphology is observed
The change of G-TOA inducing cell form as shown in Figure 3, compares normal group cell, the change of G-TOA inducing cell nuclear morphology, shoals including nuclear staining, nucleus deformation is even broken, chromatin pyknosis; Along with dosage increases, cell debris increases. Painted shoal including the forfeiture of iuntercellular contact, core, karyolysis, the increasing of cell debris; When administration concentration is 2 ��Ms and 4 ��Ms, it does not have the nucleus of intact form.
Conclusion
G-TOA shows the activity better suppressing tumor cell proliferation, and hepatoma carcinoma cell especially has good inhibitory activity; G-TOA can inducing cell karyorhexis and then cause apoptosis. Meanwhile, G-TOA has the hydrophilic being better than raw material TOA and the nephrotoxicity lower than CDDP. Therefore, this compounds can be used for the research of antitumor drug.
Detailed description of the invention
Embodiment 1 compound 2-Australia's acute pyogenic infection of nails base-3, the preparation of 5,6-trimethylpyrazines
By " " intermediate 2-Australia methyl-3, the optimization of synthesis of 5,6-trimethylpyrazines ". thread expands, Wang Penglong, Han Qiujun, etc. Anhui medicine, 2013,17 (9): 1467-1470 " prepared by method. Weigh 20.00g (0.15mol) anhydrous ligustrazine, 23.54gNBSN-bromo-succinimide (0.15mol, with front finely ground) is placed in 250mL three-necked bottle, 100mLCCl4As reaction dissolvent, 4 85W electric filament lamp irradiate, 95 DEG C of back flow reaction 1h. TLC [V (petroleum ether): V (acetone)=3: 1] detection reaction is substantially completely; Filter after cooling, collect filtrate, decompression and solvent recovery, obtain aubergine viscous liquid (content by 60%). HRMS (ESI) m/z:216.00135 [M+H]+, Theoretical Calculation: C8H11BrN2: 216.00851.
The preparation of embodiment 2 compound G-TOA
Weigh 66.57mg (0.38mmol) Boc-L-glycine successively, 95.50mg (0.50mmol) EDCI, 3.05mg (0.025mmol) DMAP and 147.75mg (0.25mmol) TOA is placed in 50mL single port flask, adding appropriate dichloromethane makes it dissolve, and stirs overnight under room temperature. TLC [V (dichloromethane): V (methanol)=20: 1] detects reaction process, reactant liquor dichloromethane extraction, collected organic layer, appropriate anhydrous sodium sulfate dewaters, evaporated under reduced pressure, silicagel column separates [V (dichloromethane): V (methanol)=20: 1] and obtains white solid; It is dissolved in the ethyl acetate containing 3MHCl, cryosel bath stirring 1h; Evaporated under reduced pressure reactant liquor, appropriate ethyl acetate is redissolved, adjust pH to neutral with saturated sodium bicarbonate solution, collected organic layer, appropriate anhydrous sodium sulfate dewaters, evaporated under reduced pressure, and silicagel column separates [V (dichloromethane): V (methanol)=20: 1] and obtains white solid, fusing point: 96.0-97.6 DEG C (c1.0, methanol), yield: 64%.1H-NMR(CDCl3) �� (ppm):1HNMR(CDCl3) �� (ppm): 0.55,0.85,0.86,0.89,1.11 (s, 15H, 5 ��-CH 3, the methyl of oleanolic acid), 0.90 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 2.24 (brs, 2H ,-NH 2), 2.49,2.51,2.55 (s, each, 3H, 3 ��-CH 3, the methyl of ligustrazine), 2.86 (m, 1H ,-CH=CCH-), 3.48 (s, 2H ,-NH2CH 2-), 4.55 (m, 1H ,-OCOCH-), 5.12,5.19 (d, each, J=10Hz, 1H ,-OCH 2-), 5.24 (brs, 1H ,-CH=C-), 1.00-2.50 (23H, the methine of oleanolic acid, methylene);13C-NMR(CDCl3) �� (ppm): 15.7,16.8,16.9,18.2,23.0,23.4,23.6,25.8,27.6,28.1,29.7,30.7,32.4,32.6,33.1,33.8,36.9,37.8,38.1,39.2,41.3,41.6,43.7 (-CH2NH2), 45.9,47.5,55.3,64.9,81.9 (-OCOCH-), 122.3 (-CH=C-), 143.6 (-CH=C-), 173.4 (-CO-), 177.2 (-CO-), pyrazine ring: 20.6 (-CH3), 21.4 (-CH3), 21.7 (-CH3), 64.9 (-COCH2-), 145.4,148.8,149.2,151.0.HRMS (ESI) m/z:648.47410 [M+H]+, Theoretical Calculation C40H62N3O4: 648.4662.
The preparation of embodiment 3 compound A-TOA
Weigh 71.90mg (0.38mmol) Boc-L-alanine successively, 95.50mg (0.50mmol) EDCI, 3.05mg (0.025mmol) DMAP and 147.75mg (0.25mmol) TOA is placed in 50mL single port flask, adding appropriate dichloromethane makes it dissolve, and stirs overnight under room temperature. Operate according to embodiment 2, obtain white solid, fusing point 89.7-92.1 DEG C,(c1.0, methanol), yield: 56%.1H-NMR(CDCl3) �� (ppm): 0.54,0.89,1.11 (s, each, 3H, 3 ��-CH 3, the methyl of oleanolic acid), 0.87 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 0.91 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 2.49,2.51,2.55 (s, each, 3H, 3 ��-CH 3, the methyl of ligustrazine), 2.74 (brs, 2H ,-NH 2), 2.86 (m, 1H ,-CH=CCH-), 3.58 (s, 1H ,-NH2CH-), 4.53 (m, 1H ,-OCOCH-), 5.12,5.19 (d, each, J=10Hz, 1H ,-OCH 2-), 5.24 (brs, 1H ,-CH=C-), 1.00-2.50 (25H, the methyl of alanine, the methine of oleanolic acid, methylene);13C-NMR(CDCl3) �� (ppm): 15.7,16.7,16.8,18.2,23.0,23.5,23.7,25.8,27.6,28.1,28.2,30.7,32.4,32.6,33.1,33.8,36.9,37.8,38.0,38.1,39.2,41.3,41.6,45.9,46.9,47.5,55.3,55.4 (-CHNH2), 81.9 (-OCOCH-), 122.3 (-CH=C-), 143.6 (-CH=C-), 173.4 (-CO-), 177.2 (-CO-), pyrazine ring: 20.6 (-CH3), 21.4 (-CH3), 21.7 (-CH3), 64.9 (-COCH2-), 145.4,148.8,149.2,151.0.HRMS (ESI) m/z:662.48967 [M+H]+, Theoretical Calculation C41H64N3O4: 662.48186.
The preparation of embodiment 4 compound P-TOA
Weigh 81.80mg (0.38mmol) Boc-L-proline successively, 95.50mg (0.50mmol) EDCI, 3.05mg (0.025mmol) DMAP and 147.75mg (0.25mmol) TOA is placed in 50mL single port flask, adding appropriate dichloromethane makes it dissolve, and stirs overnight under room temperature. Operate according to embodiment 2, obtain white solid, fusing point 89.6-91.3 DEG C,(c1.0, methanol), yield: 51%.1H-NMR(CDCl3) �� (ppm): 0.54,0.86,0.87,0.89,1.11 (s, each, 3H, 5 ��-CH 3, the methyl of oleanolic acid), 0.91 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 2.49,2.51,2.55 (s, each, 3H, 3 ��-CH 3, the methyl of ligustrazine), 2.86 (m, 1H ,-CH=CCH-), 2.95,3.10 (m, 2H ,-NHCH 2-), 3.74 (brs, 1H ,-NH-), 3.81 (m, 1H ,-NHCH-), 4.55 (m, 1H ,-OCOCH-), 5.12,5.19 (d, each, J=10.0Hz, 1H ,-OCH 2-), 5.24 (brs, 1H ,-CH=C-), 1.00-2.50 (26H, the methylene of proline, the methine of oleanolic acid, methylene);13C-NMR(CDCl3) �� (ppm): 15.7,16.9,18.1,23.0,23.4,23.6,23.7,25.4,25.8,27.6,28.1,29.7,30.5,30.7,32.4,32.6,33.1,33.8,36.9,37.8,38.1,39.2,41.3,41.6,45.9,46.9,47.0 (-NHCH2-), 47.5,55.3,60.0 (-NHCH-), 81.9 (-OCOCH-), 122.3 (-CH=C-), 143.6 (-CH=C-), 174.7 (-CO-), 177.2 (-CO-), pyrazine ring: 20.6 (-CH3), 21.4 (-CH3), 21.7 (-CH3), 64.9 (-COCH2-), 145.4,148.8,149.2,151.0.HRMS (ESI) m/z:688.50525 [M+H]+, Theoretical Calculation C43H66N3O4: 688.49751.
The synthesis of embodiment 5 compound L-TOA
Weigh 87.89mg (0.38mmol) Boc-L-leucine successively, 95.50mg (0.50mmol) EDCI, 3.05mg (0.025mmol) DMAP and 147.75mg (0.25mmol) TOA is placed in 50mL single port flask, adding appropriate dichloromethane makes it dissolve, and stirs overnight under room temperature. Operate according to embodiment 2, obtain white solid, fusing point 89.6-91.6 DEG C,(c1.0, methanol), yield: 49%.1H-NMR(CDCl3) �� (ppm): 0.54,0.89,1.10 (s, each, 3H, 3 ��-CH 3, the methyl of oleanolic acid), 0.87 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 0.91 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 0.96 (m, 6H, 2 ��-CH 3, leucic methyl), 2.49,2.51,2.54 (s, each, 3H, 3 ��-CH 3, the methyl of ligustrazine), 2.86 (m, 1H ,-CH=CCH-), 3.51 (m, 1H ,-CHNH2), 4.53 (m, 1H ,-OCOCH-), 5.12,5.19 (d, each, J=10.0Hz, 1H ,-OCH 2-), 5.24 (brs, 1H ,-CH=C-), 1.00-2.50 (methine of 27H, leucine and oleanolic acid, methylene);13C-NMR(CDCl3) �� (ppm): 15.7,16.8,18.2,21.8,23.0,23.1,23.4,23.5,23.6,24.8,25.8,27.6,28.1,29.7,30.7,32.4,32.6,33.1,33.8,36.9,37.8,38.1,39.2,41.3,41.6,43.7,45.9,46.9,47.5,53.3 (-CHNH2), 55.3,81.9 (-OCOCH-), 122.3 (-CH=C-), 143.6 (-CH=C-), 173.4 (-CO-), 177.2 (-CO-), pyrazine ring: 20.6 (-CH3), 21.4 (-CH3), 21.7 (-CH3), 64.9 (-COCH2-), 145.4,148.8,149.2,151.0.HRMS (ESI) m/z:704.53694 [M+H]+, Theoretical Calculation C44H70N3O4: 704.52881.
The synthesis of embodiment 6 compound I-TOA
Weigh 87.89mg (0.38mmol) Boc-L-isoleucine successively, 95.50mg (0.50mmol) EDCI, 3.05mg (0.025mmol) DMAP and 147.75mg (0.25mmol) TOA is placed in 50mL single port flask, adding appropriate dichloromethane makes it dissolve, and stirs overnight under room temperature. Operate according to embodiment 2, obtain white solid, fusing point 89.8-90.7 DEG C,(c1.0, methanol), yield: 49%.1H-NMR(CDCl3) �� (ppm): 0.55,0.89,1.11 (s, each, 3H, 3 ��-CH 3, the methyl of oleanolic acid), 0.88 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 0.91 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 1.01 (m, 3H ,-CH 3, the methyl of isoleucine), 2.49,2.51,2.54 (s, each, 3H, 3 ��-CH 3, the methyl of ligustrazine), 2.87 (m, 1H ,-CH=CCH-), 3.42 (m, 1H ,-CHNH2), 4.53 (m, 1H ,-OCOCH-), 5.12,5.19 (d, each, J=10.0Hz, 1H ,-OCH 2-), 5.24 (brs, 1H ,-CH=C-), 1.00-2.50 (30H, the methyl of isoleucine, methine, methylene, the methine of oleanolic acid, methylene);13C-NMR(CDCl3) �� (ppm): 11.7,15.3,15.9,16.8,16.9,18.2,23.1,23.4,23.6,23.8,24.4,25.8,27.6,28.1,30.7,32.4,32.6,33.1,33.8,36.9,37.7,38.1,38.7,39.2,41.3,41.6,45.9,46.9,47.5,55.3,59.7 (-CHNH2), 81.9 (-OCOCH-), 122.3 (-CH=C-), 143.6 (-CH=C-), 173.5 (-CO-), 177.2 (-CO-), pyrazine ring: 20.6 (-CH3), 21.4 (-CH3), 21.7 (-CH3), 64.9 (-COCH2-), 145.4,148.8,149.4,151.1.HRMS (ESI) m/z:704.53668 [M+H]+, Theoretical Calculation C44H70N3O4: 704.52881.
The synthesis of embodiment 7 compound V-TOA
Weigh 82.56mg (0.38mmol) Boc-L-valine successively, 95.50mg (0.50mmol) EDCI, 3.05mg (0.025mmol) DMAP and 147.75mg (0.25mmol) TOA is placed in 50mL single port flask, adding appropriate dichloromethane makes it dissolve, and stirs overnight under room temperature. Operate according to embodiment 2, obtain white solid, fusing point 889.5-91.0 DEG C,(c1.0, methanol), yield: 57%.1H-NMR(CDCl3) �� (ppm): 0.55,0.89,1.11 (s, each, 3H, 3 ��-CH 3, the methyl of oleanolic acid), 0.88 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 0.91 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 2.49,2.51,2.54 (s, each, 3H, 3 ��-CH 3, the methyl of ligustrazine), 2.87 (m, 1H ,-CH=CCH-), 3.31 (m, 1H ,-CHMH2), 4.53 (m, 1H ,-OCOCH-), 5.12,5.19 (d, each, J=10.0Hz, 1H ,-OCH 2-), 5.24 (brs, 1H ,-CH=C-), 1.00-2.50 (31H, the methyl of valine, methylene, the methine of oleanolic acid, methylene);13C-NMR(CDCl3) �� (ppm): 15.3,16.7,16.8,16.9,18.2,19.7,23.1,23.4,23.6,23.7,25.8,27.6,28.1,30.7,31.7,32.4,32.6,33.1,33.8,36.9,37.7,38.1,39.2,41.3,41.6,45.9,46.9,47.5,55.3,60.4 (-CHNH2), 81.9 (-OCOCH-), 122.3 (-CH=C-), 143.6 (-CH=C-), 173.5 (-CO-), 177.2 (-CO-), pyrazine ring: 20.6 (-CH3), 21.4 (-CH3), 21.7 (-CH3), 64.9 (-COCH2-), 145.4,148.8,149.4,151.1.HRMS (ESI) m/z:690.52101 [M+H]+, Theoretical Calculation C43H68N3O4: 690.51316.
The synthesis of embodiment 8 compound K-TOA
Weigh 106.51mg (0.38mmol) Cbz-L-lysine successively, 95.50mg (0.50mmol) EDCI, 3.05mg (0.025mmol) DMAP and 147.75mg (0.25mmol) TOA is placed in 50mL single port flask, adding appropriate dichloromethane makes it dissolve, stir overnight under room temperature, TLC [V (dichloromethane): V (methanol)=20: 1] detects reaction process, reactant liquor dichloromethane extraction, collected organic layer, appropriate anhydrous sodium sulfate dewaters, evaporated under reduced pressure, silicagel column separates [V (dichloromethane): V (methanol)=20: 1] and obtains white solid, it is dissolved in the methanol containing 10%Pd/C, stirred overnight at room temperature under hydrogen shield, filter, filtrate decompression is evaporated, silicagel column separate [V (dichloromethane): V (methanol)=20: 1] white solid, fusing point: 87.5-88.2 DEG C,(c1.0, methanol), yield: 69%.1H-NMR(CDCl3) �� (ppm): 0.55,0.89,1.11 (s, each, 3H, 3 ��-CH 3, the methyl of oleanolic acid), 0.88 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 0.91 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 2.49,2.51,2.54 (s, each, 3H, 3 ��-CH 3, the methyl of ligustrazine), 2.75 (m, 2H ,-CH 2NH2), 2.87 (m, 1H ,-CH=CCH-), 3.44 (m, 1H ,-CHNH2), 4.52 (m, 1H ,-OCOCH-), 5.12,5.19 (d, each, J=10.0Hz, 1H ,-OCH 2-), 5.24 (brs, 1H ,-CH=C-), 1.00-2.50 (32H, the methylene of lysine, the methine of oleanolic acid, methylene).13C-NMR(CDCl3) �� (ppm): 15.3,16.8,16.9,18.2,22.9,23.1,23.4,23.5,23.6,25.8,27.6,28.1,30.7,31.0,32.4,32.6,33.1,33.8,34.4,36.9,37.7,38.1,39.2,41.4,41.6,41.8,45.9,46.9,47.5,54.6 (-CHNH2), 55.3,81.9 (-OCOCH-), 122.3 (-CH=C-), 143.6 (-CH=C-), 175.3 (-CO-), 177.2 (-CO-), pyrazine ring: 20.6 (-CH3), 21.4 (-CH3), 21.7 (-CH3), 64.9 (-OCOCH2-), 145.4,148.8,149.2,151.0.HRMS (ESI) m/z:719.54488 [M+H]+, Theoretical Calculation C44H71N4O4: 719.53971.
The synthesis of embodiment 9 compound F-TOA
Weigh 113.75mg (0.38mmol) Cbz-L-phenylalanine successively, 95.50mg (0.50mmol) EDCI, 3.05mg (0.025mmol) DMAP and 147.75mg (0.25mmol) TOA is placed in 50mL single port flask, adding appropriate dichloromethane makes it dissolve, stir overnight under room temperature, operation according to embodiment 8, obtain white solid, fusing point 82.4-83.8 DEG C(c1.0, methanol), yield: 60%.1H-NMR(CDCl3) �� (ppm): 0.54,0.79,0.84,0.89,0.90,0.91,1.11 (s, each, 3H, 7 ��-CH 3, the methyl of oleanolic acid), 2.49,2.51,2.54 (s, each, 3H, 3 ��-CH 3, the methyl of ligustrazine), 2.87 (m, 1H ,-CH=CCH-), 3.16 (m, 2H ,-CH 2CHNH2), 3.76 (m, 1H ,-CHNH2), 4.53 (m, 1H ,-OCOCH-), 5.12,5.19 (d, each, J=10.0Hz, 1H ,-OCH 2-), 5.24 (brs, 1H ,-CH=C-), 7.22,7.23,7.29,7.30,7.31 (m, 5H ,-C6 H 5), 1.00-2.50 (24H, the methine of oleanolic acid, methylene);13C-NMR(CDCl3) �� (ppm): 15.3,16.8,16.9,18.2,23.1,23.4,23.5,23.6,25.9,27.6,28.1,30.7,32.4,32.6,33.1,33.8,36.9,37.7,38.1,39.2,41.0,41.3,41.7,45.9,46.9,47.5,55.3,56.1 (-CHNH2), 81.6 (-OCOCH-), 122.3 (-CH=C-), 143.6 (-CH=C-), 175.3 (-CO-), 177.2 (-CO-), phenyl ring: 126.8,128.5,128.6,129.3,129.4,137.2, pyrazine ring: 20.6 (-CH3), 21.4 (-CH3), 21.7 (-CH3), 64.9 (-OCOCH2-), 145.4,148.8,149.2,151.0.HRMS (ESI) m/z:738.52094 [M+H]+, Theoretical Calculation C47H68N3O4: 738.51316.
The synthesis of embodiment 10 compound Sa-TOA
Weigh 84.82mg (0.38mmol) Cbz-L-sarcosine successively, 95.50mg (0.50mmol) EDCI, 3.05mg (0.025mmol) DMAP and 147.75mg (0.25mmol) TOA is placed in 50mL single port flask, adding appropriate dichloromethane makes it dissolve, stir overnight under room temperature, operation according to embodiment 8, obtain white solid, fusing point 81.9-82.5 DEG C(c1.0, methanol), yield: 55%.1H-NMR(CDCl3) �� (ppm): 0.55,0.86,0.87,0.89,1.11 (s, each, 3H, 5 ��-CH 3, the methyl of oleanolic acid), 0.91 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 2.45 (s, 3H ,-NHCH 3), 2.49,2.51,2.54 (s, each, 3H, 3 ��-CH 3, the methyl of ligustrazine), 2.87 (m, 1H ,-CH=CCH-), 3.37 (s, 2H ,-CH 2NH-), 4.57 (m, 1H ,-OCOCH-), 5.12,5.19 (d, each, J=10.0Hz, 1H ,-OCH 2-), 5.24 (brs, 1H ,-CH=C-), 1.00-2.50 (23H, the methine of oleanolic acid, methylene);13C-NMR(CDCl3) �� (ppm): 15.4,16.7,16.8,18.3,23.1,23.4,23.6,23.6,25.8,27.6,28.1,30.7,32.4,32.6,33.1,33.9,36.0 (-NHCH3), 36.9,37.7,38.1,39.2,41.3,41.6,45.9,46.9,47.5,52.9 (-CH2NH-), 55.3,81.5 (-OCOCH-), 122.3 (-CH=C-), 143.6 (-CH=C-), 172.0 (-CO-), 177.2 (-CO-), pyrazine ring: 20.6 (-CH3), 21.4 (-CH3), 21.7 (-CH3), 64.9 (-OCOCH2-), 145.4,148.8,149.2,151.0.HRMS (ESI) m/z:662.48967 [M+H]+, Theoretical Calculation C41H64N3O4: 662.48186.
The synthesis of embodiment 11 compound Py-TOA
Weigh 49.06mg (0.38mmol) L-Glutimic acid successively, 95.50mg (0.50mmol) EDCI, 3.05mg (0.025mmol) DMAP and 147.75mg (0.25mmol) TOA is placed in 50mL single port flask, adding appropriate dichloromethane makes it dissolve, stir overnight under room temperature, TLC [V (dichloromethane): V (methanol)=20: 1] detects reaction process, reactant liquor dichloromethane extraction, collected organic layer, appropriate anhydrous sodium sulfate dewaters, evaporated under reduced pressure, silicagel column separates [V (dichloromethane): V (methanol)=20: 1] and obtains white solid, fusing point 124.6-125.8 DEG C,(c1.0, methanol), yield: 45%.1H-NMR(CDCl3) �� (ppm): 0.55,0.89,1.11 (s, each, 3H, 3 ��-CH 3, the methyl of oleanolic acid), 0.88 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 0.91 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 2.50,2.52,2.56 (s, each, 3H, 3 ��-CH 3, the methyl of ligustrazine), 2.87 (m, 1H ,-CH=CCH-), 4.25 (m, 1H ,-CHNH-), 4.57 (m, 1H ,-OCOCH-), 5.12,5.19 (d, each, J=10.0Hz, 1H ,-OCH 2-), 5.24 (brs, 1H ,-CH=C-), 1.00-2.50 (27H, the methylene of pyroglutamic acid, the methine of oleanolic acid, methylene).13C-NMR(CDCl3) �� (ppm): 15.3,16.8,16.8,18.2,23.1,23.4,23.5,23.8,25.1,25.9,27.7,28.1,29.3,30.7,32.4,32.6,33.1,33.9,36.9,37.9,38.1,39.2,41.3,41.9,45.9,46.9,47.5,55.3,55.7 (-CHNH-), 82.5 (-OCOCH-), 122.3 (-CH=C-), 143.6 (-CH=C-), 171.7 (-CO-), 177.2 (-CO-), 177.7 (-CONH-), pyrazine ring: 20.6 (-CH3), 21.4 (-CH3), 21.7 (-CH3), 64.9 (-OCOCH2-), 145.4,148.8,149.2,150.9.HRMS (ESI) m/z:702.48461 [M+H]+, Theoretical Calculation C43H64N3O5: 702.47677.
The synthesis of embodiment 12 compound W-TOA
Weigh 128.58mg (0.38mmol) Cbz-L-sarcosine successively, 95.50mg (0.50mmol) EDCI, 3.05mg (0.025mmol) DMAP and 147.75mg (0.25mmol) TOA is placed in 50mL single port flask, adding appropriate dichloromethane makes it dissolve, stir overnight under room temperature, operation according to embodiment 8, obtain white solid, fusing point 206.9-207.7 DEG C(c1.0, methanol), yield: 43%.1H-NMR(DMSO-d6) �� (ppm): 0.35,0.60,0.69,0.81,1.06 (s, each, 3H, 5 ��-CH 3, the methyl of oleanolic acid), 0.87 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 2.40,2.42,2.46 (s, each, 3H, 3 ��-CH 3, the methyl of ligustrazine), 2.76 (m, 1H ,-CH=CCH-), 2.96,3.12 (m, 2H ,-CH 2CHNH2), 3.75 (m, 1H ,-CHNH2), 4.35 (m, 1H ,-OCOCH-), 5.09,5.10, (d, each, J=10.0Hz, 1H ,-OCH 2-), 5.11 (brs, 1H ,-CH=C-), indole ring: 6.97,7.06 (m, each, 1H), 7.14 (brs ,-CHNH-), 7.34,7.53 (d, each, J=10.0Hz, 1H), 10.9 (s, 1H ,-NH), 1.00-2.50 (24H, the methine of oleanolic acid, methylene);13C-NMR(DMSO-d6) �� (ppm): 14.9,16.3,16.4,17.6,22.6,22.8,23.0,23.3,25.4,26.9,27.4,30.1,30.3,32.0,32.1,32.7,33.1,36.3,37.1,37.4,38.7,40.9,41.2,45.4,46.2,46.7,54.4,54.9 (-CHNH2), 80.6 (-OCOCH-), 121.8 (-CH=C-), 143.2 (-CH=C-), 173.3 (-CO-), 176.1 (-CO-), indole ring: 109.2,111.4,118.2,118.3,120.9,123.8,127.1,136.2, pyrazine ring: 20.1 (-CH3), 20.9 (-CH3), 21.2 (-CH3), 64.2 (-OCOCH2-), 145.0,148.2,148.7,150.6.HRMS (ESI) m/z:777.53162 [M+H]+, Theoretical Calculation C49H69N4O4: 777.52406.
The synthesis of embodiment 13 compound S-TOA
Weigh 239.22mg (1.0mmol) Cbz-L-serine successively, 180.00mg (1.2mmol) TBDMS-Cl, 136.16mg (2.0mmol) imidazoles is placed in 50mL single port flask, adding appropriate DMF makes it dissolve, stirred overnight at room temperature, TLC [V (dichloromethane): V (methanol)=20: 1] detects reaction process, reactant liquor is extracted with ethyl acetate, collected organic layer, appropriate anhydrous sodium sulfate dewaters, evaporated under reduced pressure, silicagel column separates [V (dichloromethane): V (methanol)=20: 1] and obtains intermediate; Take 83.35mg (0.38mmol) intermediate, 95.50mg (0.50mmol) EDCI, 3.05mg (0.025mmol) DMAP and 147.75mg (0.25mmol) the TOA appropriate dichloromethane of addition makes it dissolve, with dichloromethane extraction, collected organic layer, appropriate anhydrous sodium sulfate dewaters, evaporated under reduced pressure, silicagel column separates [V (dichloromethane): V (methanol)=20: 1] and obtains white solid; It is dissolved in appropriate THF, add the TBAF of 5 times amount, cryosel bath stirring 0.5h, TLC [V (dichloromethane): V (methanol)=20: 1] detects reaction process, reactant liquor dichloromethane extraction, collected organic layer, appropriate anhydrous sodium sulfate dewaters, evaporated under reduced pressure, silicagel column separates [V (dichloromethane): V (methanol)=20: 1] and obtains white solid; It is dissolved in the methanol containing 10%Pd/C, stirred overnight at room temperature under hydrogen shield; Filter, filtrate decompression is evaporated, silicagel column separate [V (dichloromethane): V (methanol)=20: 1] white solid, fusing point: 107.1-108.5 DEG C,(c1.0, methanol), yield: 36%.1H-NMR(CDCl3) �� (ppm): 0.55,0.89,1.11 (s, each, 3H, 3 ��-CH 3), 0.87 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 0.91 (brs, 6H, 2 ��-CH 3, the methyl of oleanolic acid), 2.17 (s, 1H ,-OH), 2.49,2.51,2.54 (s, each, 3H, 3 ��-CH 3, the methyl of ligustrazine), 2.71 (m, 2H ,-CH2OH), 2.87 (m, 1H ,-CH=CCH-), 4.54 (m, 1H ,-OCOCH-), 5.12,5.19 (d, each, J=10.0Hz, 1H ,-OCH 2-), 5.24 (brs, 1H ,-CH=C-), 1.00-2.50 (25H, the methine of oleanolic acid, methylene).13C-NMR(CDCl3) �� (ppm): 15.7,16.8,16.9,18.2,23.0,23.5,23.7,23.8,25.8,27.6,28.1,30.7,32.4,32.6,33.1,33.8,36.9,37.8,38.0,39.2,41.3,41.6,45.9,46.9,47.5,55.3,56.1 (-CHNH2), 63.7 (-CH2OH), 81.9 (-OCOCH-), 122.3 (-CH=C-), 143.6 (-CH=C-), 173.4 (-CO-), 177.2 (-CO-), pyrazine ring: 20.6 (-CH3), 21.4 (-CH3), 21.7 (-CH3), 64.9 (-OCOCH2-), 145.4,148.8,149.2,151.0.HRMS (ESI) m/z:678.48175 [M+H]+, Theoretical Calculation C41H64N3O5: 678.47677.
Embodiment 14
Take X-TOA10g, add tablet (including slow-release tablet, matrix tablet, coated tablet, dispersible tablet etc.) suitably adjuvant, prepare into antineoplastic agent tablet by tablet (including slow-release tablet, matrix tablet, coated tablet, dispersible tablet etc.) technique.
Embodiment 15
Take X-TOA10g, add the suitable adjuvant of capsule, prepare into antineoplastic agent capsule by capsule technique.
Embodiment 16
Take X-TOA10g, add the suitable adjuvant of granule, prepare into antineoplastic agent granule by granule technique.
Embodiment 17
Take X-TOA10g, add the suitable adjuvant of oral liquid, prepare into antineoplastic agent oral liquid by oral liquid technique.
Embodiment 18
Take X-TOA10g, add the suitable adjuvant of granule, prepare into antineoplastic agent pill by granule technique.
Embodiment 19
Take X-TOA10g, add injection (including injection, lyophilized injectable powder and aseptic subpackaged dry powder injection) suitably adjuvant, prepare into antineoplastic agent injection by injection (including injection, lyophilized injectable powder and aseptic subpackaged dry powder injection) technique.
Embodiment 20
Take X-TOA10g, add Emulsion (including microemulsion, nano-emulsion etc.) suitably adjuvant, prepare into antincoplastic agents by Emulsion (including microemulsion, nano-emulsion etc.) technique.
Embodiment 21
Take X-TOA10g, add the suitable adjuvant of Lipidosome, prepare into antineoplastic agent Lipidosome by liposome technique.
Accompanying drawing 1Giemsa staining observes the present composition G-TOA impact on HepG2 cellular morphology;
Accompanying drawing 2Giemsa staining observes the present composition G-TOA impact on mdck cell form;
Accompanying drawing 3DAPI staining observes the present composition G-TOA impact on HepG2 karyomorphism.
Claims (9)
1. there is the compounds X-TOA of below formula
2. compound as claimed in claim 1, wherein, R is nonpolar amino acid, polar amino acid and basic amino acid.
3. compound as claimed in claim 1 or 2, wherein, aminoacid is glycine, alanine, proline, leucine, isoleucine, valine, lysine, phenylalanine, sarcosine, pyroglutamic acid, tryptophan and serine.
4. the numbering of compound as claimed in claim 3 and structural formula are as follows:
5. the preparation method of compound as claimed in claim 1 or 2, including following three kinds of technology paths:
(1) TOA being dissolved in organic solvent, with the condensation under condensing agent, catalysts conditions of Boc-aminoacid, then in acid condition, de-Boc-protects base to generate G-TOA, A-TOA, P-TOA, L-TOA, I-TOA, V-TOA;
(2) TOA is dissolved in organic solvent, with the condensation under condensing agent, catalysts conditions of Cbz-aminoacid, then Pd/C-H2Under condition, de-Cbz-protects base to generate K-TOA, F-TOA, Sa-TOA, Py-TOA, W-TOA;
(3) TOA is dissolved in organic solvent, with TBDMS-Cl protection the condensation under condensing agent, catalysts conditions of Cbz-L-serine, then with TBAF take off TBDMS-protect base, last Pd/C-H2Under condition, de-Cbz-protects base to generate S-TOA.
6. the preparation method of compound as claimed in claim 5, it is characterised in that the method is:
Described reaction carries out in the mixed solvent of dichloromethane, ethyl acetate, methanol or their various ratios; Relating to condensation reaction to carry out at 20-30 DEG C, catalyst is DMAP (DMAP), and condensing agent is 1-ethyl-3-(3-dimethylamine propyl) carbodiimide hydrochloride (EDCI); Boc-protects base deprotection to carry out in-10-0 DEG C; Cbz-protects base deprotection to carry out at 20-30 DEG C; The protection of the hydroxyl of Cbz-L-serine is carrying out at 20-30 DEG C, and deprotection carries out in-10-0 DEG C; It addition, in acid used, mineral acid is hydrochloric acid.
7. compound application in preparing antitumor drug as claimed in claim 1 or 2.
8. apply as claimed in claim 7, it is characterised in that the application in preparation treatment hepatocarcinoma, gastric cancer, colon cancer, cervical cancer medicine of the described compound.
9. an antitumor medicine composition, it is characterised in that said composition comprises the compound or its salt of claim 1 or 2 and pharmaceutically acceptable carrier.
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|---|---|---|---|---|
| CN107474097A (en) * | 2016-06-07 | 2017-12-15 | 雷海民 | With antitumor action enoxolone cinnamic acid derivative（GA‑CA）Preparation method and applications |
| CN108456239A (en) * | 2017-02-20 | 2018-08-28 | 雷鹏程 | Compound BA-X with antitumor action and its preparation method and application |
| CN110964078A (en) * | 2018-09-28 | 2020-04-07 | 薪火炙药(北京)科技有限公司 | Hederagenin compound H-X with anti-lung cancer effect and preparation method and application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107474097A (en) * | 2016-06-07 | 2017-12-15 | 雷海民 | With antitumor action enoxolone cinnamic acid derivative（GA‑CA）Preparation method and applications |
| CN108456239A (en) * | 2017-02-20 | 2018-08-28 | 雷鹏程 | Compound BA-X with antitumor action and its preparation method and application |
| CN110964078A (en) * | 2018-09-28 | 2020-04-07 | 薪火炙药(北京)科技有限公司 | Hederagenin compound H-X with anti-lung cancer effect and preparation method and application thereof |
| CN110964078B (en) * | 2018-09-28 | 2021-03-23 | 薪火炙药(北京)科技有限公司 | Hederagenin compound H-X with anti-lung cancer effect and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105622704B (en) | 2018-09-28 |
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