CN104557909A - 3-acyloxy substituted (+)-deoxytylophorinine derivatives, and preparation method, pharmaceutical composition and application thereof - Google Patents

Abstract

The present invention relates to compounds represented by general formula (I), preparation methods of such compounds, pharmaceutical compositions containing them and application of such compounds in the preparation of drugs for preventing and/or treating cancer and/or inflammatory diseases.

Description

3-Acyloxy substituted D-deoxysaltenin derivatives, its preparation method, pharmaceutical composition and use

technical field

The present invention relates to compounds represented by general formula (I), preparation methods of such compounds, pharmaceutical compositions containing them and application of such compounds in the preparation of drugs for preventing and/or treating cancer and/or inflammatory diseases.

Background technique

The phenanthrene and indolizidine alkaloids are mainly distributed in the plants of the genus Romoco, and have a variety of pharmacological activities, among which the antitumor and anti-inflammatory effects are of concern. In the anti-tumor screening of the National Cancer Institute (NCI), it was found that this type of alkaloid has a significant effect on 60 tumor cell lines, and the half growth inhibitory dose (GI ₅₀ ) is at the level of 10 ^-8 M, and for malignant tumors, For example, melanoma and lung cancer cells have good selectivity, are effective for drug-resistant cancer cell lines, and have no cross-resistance with other anticancer drugs.

Among them, D-desoxysertenin was isolated from Tylophora atrofoculata and Tylophoro ovata, and after pharmacological activity screening, it was found that the screened compounds were effective for various Tumor cells from different tissues, including human glioblastoma U251 and BT323 cell lines, have significant activity, IC ₅₀ is at the level of 10 ^-8 -10 ^-7 M, and the in vivo test results show that it can inhibit the growth of mouse H22 xenografts The inhibition rate is 70-80%. Previous studies on the mechanism of action indicated that this compound may function by intercalating between the AT base pairs of DNA and RNA, which is different from clinical antitumor drugs [Chemotherapy.2011,57:310-320].

Through drug metabolism research, it was found that the main metabolite of this compound in experimental animals is 3-desmethyl-D-desoxysaltenin [Anal.Chim.Acta.2012,731,60], which was synthesized and screened It was found that 3-desmethyl-D-desoxysaltenine has very significant cytotoxic activity [Synthesis.2012,44,A-H], this patent involves structural optimization of this compound to improve its physical and chemical properties and in vivo activity work.

Contents of the invention

The technical problem to be solved by the present invention is to provide the compound of general formula (I) and its pharmacodynamically acceptable salt.

Another technical problem to be solved by the present invention is to provide the preparation of the compound of general formula (I) and its pharmacodynamically acceptable salt.

Another technical problem to be solved by the present invention is to provide a pharmaceutical composition comprising at least one compound of general formula (I) and a pharmacodynamically acceptable salt thereof.

Another technical problem to be solved by the present invention is to provide the compound of general formula (I) and its pharmacodynamically acceptable salts in the preparation of drugs for the prevention and/or treatment of cancer and/or inflammation.

In order to solve the problems of the technologies described above, the technical solution adopted in the present invention is:

The present invention relates to compounds represented by general formula (I):

Wherein, R is selected from R ₁ CO- or R ₂ SO ₂ -;

When R is selected from R ₁ CO-, R ₁ is selected from substituted or unsubstituted C ₁₁₀ straight chain and branched chain alkyl, substituted or unsubstituted C ₁₁₀ straight chain and branched alkoxy, substituted or unsubstituted Substituted amine groups forming five-membered, six-membered or seven-membered rings;

When R is selected from R ₂ SO ₂ -, R ₂ is selected from substituted or unsubstituted C ₁₁₀ alkyl and phenyl.

Preferred compounds according to the invention include:

According to the present invention, the compound of the present invention may exist in the form of isomers, and generally speaking "the compound of the present invention" includes the isomers of the compound.

According to an embodiment of the present invention, the compound of the present invention also includes a pharmacodynamically acceptable salt thereof.

Also provided in accordance with the present invention are processes for the preparation of compounds of the present invention.

The present invention also relates to a pharmaceutical composition containing a pharmaceutically effective dose of the compound as described in general formula I and a pharmaceutically acceptable carrier.

Still another aspect of the present invention relates to a pharmaceutical composition comprising the compound of the present invention as an active ingredient. The pharmaceutical composition can be prepared according to methods known in the art. Any dosage form suitable for human or animal use can be prepared by combining the compounds of the present invention with one or more pharmaceutically acceptable solid or liquid excipients and/or adjuvants. The content of the compound of the present invention in its pharmaceutical composition is usually 0.1-95% by weight.

The compound of the present invention or the pharmaceutical composition containing it can be administered in the form of a unit dosage, and the route of administration can be enteral or parenteral, such as oral, intravenous injection, intramuscular injection, subcutaneous injection, nasal cavity, oral mucosa, eye, lung and Respiratory tract, skin, vagina, rectum, etc.

The dosage form for administration may be a liquid dosage form, a solid dosage form or a semi-solid dosage form. Liquid dosage form can be solution (including true solution and colloid solution), emulsion (including o/w type, w/o type and double emulsion), suspension, injection (including water injection, powder injection and infusion), eye drops formulations, nasal drops, lotions and liniments, etc.; solid dosage forms can be tablets (including ordinary tablets, enteric-coated tablets, buccal tablets, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets), capsules ( Including hard capsules, soft capsules, enteric-coated capsules), granules, powders, pellets, dripping pills, suppositories, films, patches, gas (powder) mist, sprays, etc.; semi-solid dosage forms can be ointment, Gels, pastes, etc.

The compound of the present invention can be made into common preparations, sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle drug delivery systems.

To form the compound of the present invention into tablets, various excipients known in the art can be widely used, including diluents, binders, wetting agents, disintegrants, lubricants, glidants. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, iso Propanol, etc.; binders can be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, arabic mucilage, gelatin slurry, sodium carboxymethylcellulose, methylcellulose, hypromellose Base cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked poly Vinylpyrrolidone, croscarmellose sodium, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitan fatty acid ester, sodium dodecylsulfonate, etc.; lubricant and flow aid The agent can be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol and the like.

Tablets can also be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.

In order to make the administration unit into a capsule, the active ingredient compound of the present invention can be mixed with a diluent and a glidant, and the mixture can be directly placed in a hard capsule or a soft capsule. The active ingredient compound of the present invention can also be made into granules or pellets with diluents, binders, and disintegrating agents, and then placed in hard capsules or soft capsules. Various diluents, binders, wetting agents, disintegrants, and glidants used in the preparation of tablets of the compound of the present invention can also be used in the preparation of capsules of the compound of the present invention.

In order to make the compound of the present invention into injection, water, ethanol, isopropanol, propylene glycol or their mixtures can be used as solvent and an appropriate amount of commonly used solubilizers, cosolvents, pH regulators and osmotic pressure regulators in this field can be added. The solubilizer or co-solvent can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; the pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be Sodium chloride, mannitol, glucose, phosphate, acetate, etc. For preparation of freeze-dried powder injection, mannitol, glucose, etc. can also be added as proppants.

In addition, coloring agents, preservatives, fragrances, flavoring agents or other additives can also be added to the pharmaceutical preparations, if necessary.

In order to achieve the purpose of medication and enhance the therapeutic effect, the medicine or pharmaceutical composition of the present invention can be administered by any known administration method.

The dosage of the pharmaceutical composition of the compound of the present invention can vary widely depending on the nature and severity of the disease to be prevented or treated, individual conditions of the patient or animal, administration route and dosage form, etc. Generally speaking, the suitable daily dosage range of the compound of the present invention is 0.001-150 mg/Kg body weight, preferably 0.1-100 mg/Kg body weight, more preferably 1-60 mg/Kg body weight, most preferably 2-30 mg/Kg body weight. The above-mentioned dosage can be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the doctor and the dosage regimen including the use of other therapeutic means.

The compound or composition of the present invention can be taken alone, or used in combination with other therapeutic drugs or symptomatic drugs. When the compound of the present invention has a synergistic effect with other therapeutic drugs, its dose should be adjusted according to the actual situation.

The present invention also relates to the application of the compounds of the present invention in the preparation of drugs for the treatment of cancer and/or inflammatory diseases, especially in human glioma, human bone marrow neuroblastoma, human colon cancer, human gastric cancer, human ovarian cancer, Application in cervical cancer, liver cancer, lung cancer, pancreatic cancer and other cancers.

Description of drawings

Figure 1 Animal body weight records in each group

Figure 2 Records of animal tumor weight in each group

Detailed ways

The starting compounds used in the examples of the present invention can be prepared according to conventional methods in the art and/or methods well known to those skilled in the art, and can be prepared according to the following preparation examples.

The following examples are used to further illustrate the present invention, but this does not imply any limitation to the present invention.

Example 1: Synthesis of (13aS)-3-benzyloxyacyloxy-6,7-dimethoxy-9-phenanthrene[9,10-b]-indolizidine (1)

Add 50 mL of dichloromethane to a 100 mL dry reaction bottle, dissolve 100 mg of the reactant 3-hydroxy-6,7-dimethoxyphenanthrene-9-methanol (PF403), add 60 μL of triethylamine, and then add chloroformic acid 49 μL of benzyl ester was reacted for 2 hours, the reaction solution was washed with saturated ammonium chloride, dried, evaporated to dryness and separated by silica gel column chromatography, the eluent was dichloromethane/methanol (50/1), and the product 0.11g was obtained, the yield 79%. ¹ H-NMR(500MHz,C ₅ D ₅ N):8.93(1H,d,J=2.5Hz),7.65(1H,dd,J=9.0Hz,2.5Hz),8.07(1H,d,J=9.0 Hz),8.29(1H,s),7.44(1H,s),7.53(2H,m),7.36(3H,m),3.95(3H,s,MeO),3.94(3H,s,MeO),5.41 (2H,s),4.81(1H,d,J=15.0Hz),3.69(1H,d,J=15.0Hz),3.35(2H,m),2.93(1H,m),2.40(1H,m) ,2.31(1H,m),2.08(1H,m),1.87(1H,m),1.71(2H,m).

Example 2: Synthesis of (13aS)-3-ethoxyloxy-6,7-dimethoxy-9-phenanthrene[9,10-b]-indolizidine (2)

Add 50 mL of dichloromethane to a 100 mL dry reaction bottle, dissolve 100 mg of the reactant 3-hydroxy-6,7-dimethoxyphenanthrene-9-methanol (PF403), add 60 μL of triethylamine, and then add chloroformic acid 33 μL of ethyl ester was reacted for 2 hours, the reaction solution was washed with saturated ammonium chloride, dried, evaporated to dryness and separated by silica gel column chromatography, the eluent was dichloromethane/methanol (50/1), and the product was 0.09g, the yield 75%. ¹ H-NMR(500MHz,C ₅ D ₅ N):8.92(1H,d,J=2.0Hz),7.67(1H,dd,J=9.0Hz,2.0Hz),8.06(1H,d,J=9.0 Hz),8.28(1H,s),7.41(1H,s),3.96(3H,s,MeO),3.94(3H,s,MeO),4.78(1H,d,J=15.0Hz),3.67(1H ,d,J=15.0Hz),4.29(2H,q),1.22(3H,t),3.35(2H,m),2.92(1H,m), 2.38(1H,m),2.31(1H,m) ,2.08(1H,m),1.87(1H,m),1.71(2H,m).

Example 3: Synthesis of (13aS)-3-pivaloyloxy-6,7-dimethoxy-9-phenanthrene[9,10-b]-indolizidine (3)

Add 50 mL of dichloromethane to a 100 mL dry reaction bottle, dissolve 100 mg of the reactant 3-hydroxy-6,7-dimethoxyphenanthrene-9-methanol (PF403), add 60 μL of triethylamine, and then add neopentyl Acyl chloride 42 μL, reacted for 2 hours, the reaction solution was washed with saturated ammonium chloride, dried, evaporated to dryness and separated by silica gel column chromatography, eluent dichloromethane/methanol (50/1), the product 0.09g was obtained, the yield was 72 %. ¹ H-NMR(500MHz,C ₅ D ₅ N):8.96(1H,d,J=1.5Hz),7.60(1H,dd,J=8.5Hz,1.5Hz),8.14(1H,d,J=8.5 Hz),8.36(1H,s),7.43(1H,s),3.92(3H,s,MeO),3.85(3H,s,MeO),4.81(1H,d,J=15.0Hz),3.69(1H ,d,J=15.0Hz),3.35(2H,m),2.94(1H,m),2.40(1H,m),2.33(1H,m),2.08(1H,m),1.87(1H,m) ,1.69(2H,m), 1.35(9H,s).

Example 4: Synthesis of (13aS)-3-methanesulfonyloxy-6,7-dimethoxy-9-phenanthrene[9,10-b]-indolizidine (4)

Add 50 mL of dichloromethane to a 100 mL dry reaction bottle, dissolve 100 mg of the reactant 3-hydroxy-6,7-dimethoxyphenanthrene-9-methanol (PF403), add 60 μL of triethylamine, and then add neopentyl Acyl chloride 27 μL, reacted for 2 hours, the reaction solution was washed with saturated ammonium chloride, dried, evaporated to dryness and separated by silica gel column chromatography, the eluent was dichloromethane/methanol (50/1), and the product was obtained. 0.08g, yield 66 %. ¹ H-NMR(500MHz,C ₅ D ₅ N):8.96(1H,d,J=2.0Hz),7.78(1H,dd,J=9.0Hz, 2.0Hz),8.06(1H,d,J=9.0 Hz),8.26(1H,s),7.43(1H,s),3.94(3H,s,MeO),3.90(3H,s,MeO),4.84(1H,d,J=15.0Hz),3.74(1H ,d,J=15.0Hz),3.53(3H,s),3.36(2H,m),2.97(1H,m),2.47(1H,m),2.37(1H,m),2.10(1H,m) , 1.91(1H,m), 1.74(2H,m).

Example 5: (13aS)-3-(4′-piperidinyl)piperidinyloxy-6,7-dimethoxy-9-phenanthrene[9,10-b]-indolizidine (5) Synthesis

Add 50 mL of dichloromethane to a 100 mL dry reaction bottle, dissolve 100 mg of the reactant 3-hydroxy-6,7-dimethoxyphenanthrene-9-methanol (PF403), add 60 μL of triethylamine, and then add neopentyl Acyl chloride 90 mg, reacted for 2 hours, the reaction solution was washed with saturated ammonium chloride, dried, evaporated to dryness and separated by silica gel column chromatography, the eluent was dichloromethane/methanol (50/1), and the product was obtained. 0.12g, yield 77 %. ¹ H-NMR(500MHz, C ₅ D ₅ N):8.98(1H,d,J=2.0Hz),7.73(1H,dd,J=9.0Hz,2.0Hz),8.11(1H,d,J=9.0 Hz),8.34(1H,s),7.44(1H,s),3.93(3H,s,MeO),3.88(3H,s,MeO),4.82(1H,d,J=15.0Hz),3.70(1H ,d,J=15.0Hz),4.51(1H,m),4.42(2H,m),3.49(1H,m),3.37(2H,m),2.95(1H,m),2.78(2H,m) , 2.37(5H,m), 2.09(1H,m), 1.87(1H,m), 1.70(2H,m), 1.64(1H,m), 1.49(5H,m), 1.27(4H,m).

Example 6: (13aS)-3-(2′-S-tert-butoxycarbonylamino)propionyloxy-6,7-dimethoxy-9-phenanthrene[9,10-b]-indole Synthesis of Lixidine (6)

Add 50 mL of dichloromethane to a 100 mL dry reaction bottle, dissolve 100 mg of the reactant 3-hydroxy-6,7-dimethoxyphenanthrene-9-methanol (PF403), and add Boc-protected alanine under ice bath Add 0.11 g of acid, then add 0.11 g of EDCI, react for 0.5 hours, rise to room temperature, react for another 2 hours, wash the reaction solution with water, dry, evaporate to dryness and separate by silica gel column chromatography to obtain 85 mg of the product with a yield of 57%. ¹ H-NMR(500MHz,C ₅ D ₅ N):8.83(1H,d,J=2.0Hz),7.61(1H,dd,J=8.5Hz,2.0Hz),8.05(1H,d,J=8.5 Hz),8.22(1H,s),7.40(1H,s),3.90(3H,s,MeO),3.87(3H,s,MeO),4.78(1H,d,J=14.5Hz),3.67(1H ,d,J=14.5Hz),4.90(1H,m),3.33(2H,m),2.91(1H,m),2.39(1H,m),2.30(1H,m),2.05(1H,m) , 1.85(1H,m), 1.70(1H,m), 1.66(2H,d), 1.63(1H,m), 1.52(9H,s).

pharmacological test

Experimental example 1: Determination of anti-tumor activity in vitro (MTT method)

In order to measure the in vitro antitumor activity of the compounds of the present invention, the compounds prepared in the examples of the present invention were determined, and the experimental steps are:

1. Cultivate normal growing tumor cells, inoculate 1×10 ⁴ cell/mL into 96-well plate (100 μL per well), and incubate in 37°C, 5% CO ₂ incubator for 24 hours.

2. Add the test compound respectively, and culture in a 5% CO ₂ , complete humidity incubator for 5 days.

3. Discard the culture medium, add 0.04% MTT100μL to each well, and incubate under the same conditions for 4 hours.

4. Discard the culture medium, add DMSO (150 μL per well), mix and record the light absorbance colorimetrically at the measurement wavelength of 570 nm and the reference wavelength of 450 nm, and calculate the inhibitory rate of the compound on tumor cell growth.

Table 1 The growth inhibitory effect of PF403 and its prodrugs on tumor cells cultured in vitro (72h)

HepG2: human liver cancer cells; Mia-PaCa2: human pancreatic cancer cells; SHSY-5Y: human bone marrow neuroblastoma cells; SKOV3: human ovarian cancer cells; TJ905: human glioma cells

It can be seen from the results that compounds No. 1 to No. 6 all have certain in vitro anti-tumor cell growth inhibitory effects, but they are weaker than their prototype compound PF403.

Experimental example 2: Determination of anti-tumor activity in vivo

1. Experimental animals and grouping: ICR mice, SPF grade, female, weighing 18-20g. Purchased from: Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. After inoculating G422 mouse glioblastoma in the armpit (tumor fluid: sterile saline = 1:3), the next day they were randomly divided into groups, weighed and given medicine. Randomly divided into blank control group, positive control group (cyclophosphamide), administration group (No. kg, 2.5mg/kg intraperitoneal injection group).

2. Drug preparation and administration method: Cyclophosphamide for injection is dissolved in sterile normal saline (concentration 10mg/ml, intraperitoneal injection of 0.2ml per 20g mouse) before use, and intraperitoneal injection once on the first day after inoculation . Test compounds 1, 2, 3, 4, 5, and 6 were dissolved in double-distilled water (concentration: 5 mg/ml, 2.5 mg/ml, 0.4 ml per 20 g of mice) after being dissolved with Tween 80, and administered orally every other day Administration; PF403 was solubilized with polyethylene glycol 400, dissolved in sterile normal saline (final concentration of polyethylene glycol 25%), and administered by intraperitoneal injection; the test compound and PF403 were prepared immediately and stored in the dark.

3. Animal treatment: Record the body weight every other day, and draw the body weight change curve. On the 15th day after inoculation, the animals were sacrificed by cervical dislocation, the tumor was stripped, photographed, the tumor weight was recorded and the tumor inhibition rate was calculated.

Table 2 Screening results of PF403 prodrugs for in vivo anti-tumor effects on G422 mouse glioblastoma

+: The prodrug was administered orally, administered on the 2nd, 4th, 6th, 7th, 9th, 11th, and 13th days after inoculation, and administered 7 times in total; the injection dose of PF403 was the total dose, and the high-dose group 10mg/kg on the 5th day, 15mg/kg on the 7th, 10th, and 12th day, 20mg/kg on the 13th, and 14th day, 7 times in total; The administration time was the same as that of the high-dose group, and the administration amount was half of that of the middle-dose group.

#p<0.05; ##p<0.01; ###p<0.001; *p<0.05; **p<0.01; ***p<0.001, compared with the negative control group.

It can be seen from the results that compounds No. 1 to No. 6 all have strong inhibitory activity on G422 tumors, and compounds No. 2, 3, 5 and 6 have stronger inhibitory effects than the prototype compound PF403, and the dose-effect relationship is better, and the degree of weight loss is lower than that of the prototype compound PF403. There are few other compounds.

Claims (8)

1. A compound represented by the following general formula (I), and a pharmacodynamically acceptable salt thereof: Wherein, R is selected from R ₁ CO- or R ₂ SO ₂ -; When R is selected from R ₁ CO-, R ₁ is selected from substituted or unsubstituted C ₁₁₀ straight chain and branched chain alkyl, substituted or unsubstituted C ₁₁₀ straight chain and branched alkoxy, substituted or unsubstituted Substituted amine groups forming five-membered, six-membered or seven-membered rings; When R is selected from R ₂ SO ₂ -, R ₂ is selected from substituted or unsubstituted C ₁₁₀ alkyl and phenyl. 2. according to the compound of claim 1, it is characterized in that, described compound is the compound shown in general formula (IA), and its pharmaceutically acceptable salt: Wherein, R ₁ is selected from substituted or unsubstituted C ₁₁₀ straight chain and branched chain alkyl, substituted or unsubstituted C ₁₁₀ straight chain and branched alkoxy, substituted or unsubstituted five-membered, six-membered Or the amine group of the seven-membered ring. 3. according to the compound of claim 1, it is characterized in that, described compound is the compound shown in general formula (IB), and its pharmaceutically acceptable salt: Wherein, R ₂ is selected from substituted or unsubstituted C ₁₁₀ alkyl and phenyl. 4. according to the compound of claim 1 to 3, it is characterized in that, described compound is selected from: 5. A pharmaceutical composition, characterized in that it contains an effective dose of any compound according to any one of claims 1-4 and a pharmaceutically acceptable carrier. 6. The pharmaceutical composition according to claim 5, characterized in that, the pharmaceutical composition is selected from tablets, capsules, pills, injections, sustained-release preparations, controlled-release preparations and various particle delivery systems. 7. Use of the compound according to any one of claims 1-4 in the preparation of medicines for preventing and/or treating cancer and/or inflammatory diseases. 8. according to the application of claim 7, it is characterized in that, described cancer is selected from human glioma, human bone marrow neuroblastoma, human colon cancer, human gastric cancer, human ovarian cancer, cervical cancer, liver cancer, lung cancer, pancreatic cancer.