TWI236478B - Antitumor derivative of bis-dicarboxylic acid diaminoplatin complex, and the pharmaceutical composition containing the same - Google Patents

Abstract

The present invention relates to an antitumor derivative of bis-dicarboxylic acid diaminoplatin complex of formula (I), wherein R1 and R2 have the meanings defined in the specification, also relates to a process for preparation thereof, and to an antitumor pharmaceutical composition containing the same. The composition is a new generation of safe and effective antitumor drug, which has advantages such as low toxicity, broad antitumor spectrum, high antitumor activity, good solubility in water and fine stability in aqueous solution, selective killing of cancer cells without damaging normal cells, etc.

Description

Ϊ236478 发明 Description of the invention: [Technical field to which the invention belongs] The present invention relates to a derivative of _____, which relates to a novel anti-tumor bis-diamine diamino complex; the present invention also relates to the double-two Preparation of mai diamino complex derivative and green containing the bis-diamino-charm saccharin age compound; the present invention further relates to the bis-diamino complex of the anti-drug drug Application. [Previous Technology] Since B. Rosenber discovered that cis-digas diammonium has anti-tumor properties in 1969, cisplatin (CisPlatln) has been widely used in clinical practice as a platinum-based anticancer drug. For urogenital cancer, nasopharyngeal cancer, head and neck Cancer, lung cancer and other cancers have significant curative effects, but serious toxic and side effects. Adverse reactions such as nephrotoxicity, neurotoxicity, ototoxicity, and vomiting have limited the dose and length of the drug. Carboplatin (also known as carboplatin), the second-generation anticancer drug, has anticancer spectrum and Cisplatin is similar, its efficacy is slightly inferior to cisplatin, and it has cross-resistance. Although the toxic and side effects are significantly lower than cisplatin, bone mineral suppression still exists, and its stability in water is poor. Studies on toxic and broad-spectrum platinum-based anticancer agents have been very active. The prior arts most relevant to the present invention are WO95 / 20956 and EP642792A1, both of which disclose cis-diamino-1,1-cyclobutanedi When formic acid is synthesized, its structural formula is very different from that of the bis-dicarboxylic acid diammine platinum derivative according to the present invention. Therefore, the present invention is a pioneering invention. According to reports, in recent years, there have been many new platinum-based anticancer drugs. Enter clinical trials, such as Isoplatin ^ ^^ i0 (Oxaliplatin) > ^^ l0 (Ormajplatin) > ^ e, 16 (Lobapatin) > Enloplatin, Zeniriplatin , As well as L-NDOP, DWA-2114A, CI-973, etc. Most of these new-layup compounds are cross-linked with Lin Wuxi However, the anti-tumor sharpness is roughly the same as the cis axis, there is no broad anti-cancer spectrum, and the stability in water is also poor. Most of them are lower than cis, and some still show obvious nephrotoxicity, neurotoxicity and tilt inhibition. Therefore Neither has been used in clinical applications. As for oral effective platinum anticancer drugs, no report has been reported so far. A more effective, less toxic, and stable platinum water drug is required. 1236478 The compound has undergone intensive research, and it has been unexpectedly discovered that the bi-n-junction n7 / 7 organism can overcome the above-mentioned shortcomings, has high performance, low toxicity, and stability in water, thereby achieving the present invention. [Summary of the Invention] Therefore 'The primary object of the present invention is to provide a novel antitumor diaminoplatinate derivative of bisdicarboxylic acid which can overcome the shortcomings of the prior art described above; another object of the present invention is to provide a bisdipic acid diamine Method for amoxicillin derivative; A further object of the present invention is to provide an antitumor pharmaceutical composition containing the bisdicarbodiamidine derivative as an active ingredient; the present invention The purpose of this step is to provide the application of the shuangdijian diaminoluozaki organisms in the preparation of antitumor drugs. The invention / 歩 and an antitumor bismonotonic acid diammonium derivative are characterized in that: It has the following formula (I): ^

In the formula: R1 and R2 may be the same or different, and they independently represent hydrogen, Clw hydrocarbyl, hydrogen, aminocyano, hydroxyl, carboxyl, acyl, phosphoryl, or phosphorylamino, or R1 and R2 are connected to each other; The connected carbon atoms together form a carbon atom-containing LG saturated or unsaturated carbocyclic ring. Preferably, said R1 and R2 are connected to each other and together with the carbon atom to which they are connected form a 3-6 membered saturated carbocyclic ring containing carbon atoms. 1236478 It is more preferable that the dual-secondary diamine complex is a dual-u-cyclopropane secondary-diamine complex or a bis-1,1-cyclobutanedicarboxylic acid diaminoplatinum compound. Most preferably, the present invention provides a bis-u. Cyclidine diarimidine complex compound of the following formula (hereinafter referred to as bicyclic platinum):

Said :: = :: double, two_biological method'its special 〇 ----- Η— 〇, c-〇, CHh :: / \ h, \ h 〇, c / \ r2 H-- A certain H f may be the same, which independently represents hydrogen, -hydrocarbyl, autogen, amino, lactyl, meridian, vinyl, cationic, or amino; or R and R are connected to each other and connected to them. Carbon atom one; a saturated or unsaturated carbocyclic ring, the method is: R1 R2

X: ° > \ / R1-starting to form 3-12 carbon atoms

〇 II ^ (1) Chinoic acid ligand derivative reaction ΌΗ R1

C R2〆 II 0 ΌΗ (III) 8 1236478

In the formula: the definitions of R1 and r2 are as described in the formula (i) above to form a bis-dicarboxylic acid diammine platinum derivative of the formula (I); or 2) a dihalo-diammine platinum of the formula (NH3) 2PtX2 In the formula: X is C1 or I reacted with silver lignostearate or silver sulfate in an aqueous solution to form a monoammonium platinum nitrate hydrate of (NH3) 2pt (H2〇) 2 (N〇3) 2 or (NH3) 2Pt (H20) 2 (S04) hydrated diammine platinum sulfate; reacting the hydrated diammine platinum nitrate or sulfate with a dicarboxylic acid ligand derivative of the following formula (III) or its sodium or barium salt, R1

R2

0II / d-OH \ C-OH (III) where R1 and R2 are as defined in formula (j) above to form a bisdicarboxylic acid diammine platinum derivative of formula (I). [Embodiment] In the preparation of diammonium diammonium complex of formula (I), the reaction temperature in each step can be performed in a relatively wide range. Generally, the reaction is carried out at a temperature between (^^ to 10 (rc), preferably at a temperature ranging from 50 to (rc). The reaction time of each step is generally 2 to 16 hours. The green of the present invention is usually carried out under normal pressure. However, this method of fortune can also be carried out under pressure or decompression, usually at a pressure between 0. bar and 10 bar. ≪ When carrying out the method of the present invention, generally an approximately equimolar amount is used. Starting materials. However, it is also possible to use a relatively large excess of one of the starting materials. The starting materials, bidentate diaminoplatinum and dicarboxylic acid ligand derivatives, are known substances and can be prepared by known methods. ^ In addition The present invention also relates to a kind of antitumor pharmaceutical composition, which is characterized in that the composition 1236478 contains at least one kind of bisdinuolinium derivative of formula IX in a concentration of 0 Mwt% as a test component. And a pharmaceutically acceptable carrier. According to the pharmaceutical composition of the present invention, it is preferred that the bis-diaminine derivative of formula (I) as an active ingredient is a compound of formula ⑴r1 ^ r2 Those that are attached and together with the carbon atoms to which they are attached form a 3 to 6 membered saturated carbocyclic ring containing carbon atoms (1) The diaminodiamine platinum derivative of dicarboxylic acid is more preferably dil5; u-cyclopropanedicarboxylic acid diaminoplatinum or the dicyclobutadiene diaminodiamine complex of formula (π), most preferably Preferred is the bis, 1_cyclobutane diamine diammonium compound of formula (Π). The preparation of the antitumor pharmaceutical composition of the present invention is relatively simple, as long as the bis The carboxylic acid diammine platinum derivative may be mixed with a pharmaceutically acceptable carrier, and the pharmaceutical composition may be administered in the form of an injection or capsule. The pharmaceutically acceptable carrier may include purified water, medical starch and vitamin c In which, the medicine can be made into injections with pure water, which can be injected subcutaneously, intraperitoneally, and intravenously. The medicine can be made into oral capsules with medical starch and vitamin c. The pharmaceutical composition according to the present invention, such as bicyclic platinum for intraperitoneal injection Its LD5G is 283mg / kg, its LD50 = 50.46 ± 0.93mg / kg for intravenous injection, and its LD50 is 500mg / kg for oral administration. Ld50 is i80mg / kg. The dosage of the pharmaceutical composition according to the present invention is generally one part of its Ld5g amount. The present invention further relates to the use of the diaminoplatinum diaminodicarboxylic acid derivative of formula () in the preparation of antineoplastic drugs. The diaminoplatinum diaminodicarboxylic acid derivative (containing Transition metals other than platinum, such as Rh, Ru, Pd, Cu, Zn, Cr, etc.), for example, bis, ι_cyclobutanedicarboxylic acid diammine platinum, is a new class of supramolecular cage compounds. Covalent, coordination, and hydrogen bonds are used to connect atoms in such supramolecules. It shows its structural homogeneity and stability in solid phase and aqueous solution. The hydrogen bond is defined as molecular switch. The most important thing in life is DNA. DNA is double-encapsulated with four kinds of organic salt, namely Guanine (G), Cytosine (C), Adenine (A), and 1236478 Thymine. (Thymine, T) A-T, C-G are regularly arranged by hydrogen bonds. Hydrogen bond rupture means that a lesion has occurred. The compound of the present invention enters the human body. The hydrogen bonds in this cage-like supramolecular compound are not complementary to the hydrogen bonds in normal and normal DNA. It does not open the switch and does not work. When it encounters abnormal DNA, the hydrogen bond of the compound of the present invention and Abnormal DNA (cancer-containing DNA, Gucogene) hydrogen bonds are complementary. The molecular switch of the compound is immediately opened, so that Pt (containing other transition metals) d is complexed with purines and pyrimidines. This is a scientific concept achieved by the compounds of the present invention that selectively, specifically, blocks abnormal DNA (including Oncogene) without harming the normal characteristics of human normal DNA. A number of acute toxicity, pharmacodynamic experiments have been carried out on bis-glycerol diammonium diammonium compound of formula (I), such as diammonium diammonium diammonium diammonium compound of formula (II). After several years of clinical tests, the results are as follows: Nude mice were tested by intravenous injection, intraperitoneal injection and oral acute poisoning. Acute Toxicity Limit Test® shows that the toxicity is low, LD50 (intravenous injection) = 50.46 ± 0.93 mg / Kg, LD50 (intraperitoneal injection) = 283 mg / Kg, and LD50 (oral injection) = 500 mg / Kg 〇—Chinese Medical Science New Tumor Hospital and Cancer Research Institute conducted a certain effect-related experiment on Shuanglai. Shuanghuan started on human liver cancer cells BEL-7402IC50 = l_3pg / ml (Note: kill this cancer cell-half f dual jobs 1.3pg / ml); private breast cancer cells IC5G = 2 8) Llg / ml, human breast Carcinoma, lung cancer, occupation, large secret nest and dual-duty simple age experiment, intraperitoneal injection, divided into large doses (LD50xl / 5), medium doses (LD50xl / 10) and low doses (LD50xl / 20) group, the tumor inhibition rates were 90%, 95% and 70%, respectively. Gavage with the same dose as described above, the tumor inhibition rate is slightly lower. # Beijing Institute of Drug Control for human liver cancer% nude mice vaccination rate experiment, intravenous injection, divided into large doses (LD50xl / 5), medium doses (LD50xl / 1/10) and low doses (LD50〇 / 2 〇), and the effect of melamine and melamine. Bicyclic uranium was used in large, medium, and small doses at 0.2 ml / time, once a day, and 7 consecutive injections; sacrifice He Tumor rats were sacrificed for 30 days. The antitumor rates of large, medium and small doses of Shuanghuanming were 50 Tao and 57 Na, respectively. ^ = 0% ‘Moles live normally’ and diet is normal ’None of the 10 nude mice in each group died. And the ring was mixed and the female was called the minimum dose administration only 2 times. The inhibition rate reached 73, but most of them were reduced to death. Γ ° ° Shuanghuan was injected with rabbits to perform pharmacokinetic experiments, and mice were double-turned. Do oral medicine 11 1236478 generation experiment. The content of the scale of the animal serum towel and the minute of the drug reached a peak in money, and the content in the blood tended to 0 in 24 hours. … The long-term experiment of biaxial three-axis oral administration ... To evaluate the safety of dual-hybrids, observe the three consecutive drug administrations and provide toxic anti-eunuchs and the reversibility of their damage to the critical response of the agency and its severity Compared with non-toxic reaction seams, it provides a reference for the preparation of human safety seams. 3. The drug has a dose-effect relationship to visceral damage. Under the treatment dose, the symptoms are relieved after one month of stopping the medicine. The small dose group had little effect and returned to normal. In vitro tumor suppression experiments: Growth inhibition of human liver cancer BEL_74G2 cells cultured in vitro by Shuanghuan Jing Observed the dynamic growth of human liver cancer BEL-7402 cells after treatment with bicycloplatin, and found that: Two days after treatment, the number of cells in the control group was not significantly different from that in the control group. From 3 days after treatment, the number of viable cells in the treatment group began to be significantly less than that in the control group. The control group showed logarithmic growth, while the number of cells in the bicycloplatin treatment group It gradually decreased, and the differences between the groups became more obvious with the passage of time, and the number of viable cells in the 20 pg / ml paired bicyclic platinum group was lower than the 10 μβ / πιιππ platinum group. The above results show that Shuangbaibai has a significant growth inhibitory effect on human liver BEL · · cells cultured in vitro, and shows a time-dependent effect relationship (see Figure 4). 2. Double% platinum has the function of selectively killing cancer cells within a certain dose range without harming normal cells. The Nanbuzi College of Life Sciences used Shuanghuan to conduct experiments on human normal fibers, epidermal cells, lung cancer cells, and melanoma. The experiments proved that normal human fibroblasts and epidermal cells were not damaged, and lung cancer cells and melanocytes were killed by 50% and 80% (see Figure 5). 3. Effect of bicycloplatin on the ultrastructure of human liver cancer BEL-7402 cells. After BEL-7402 cells were treated with bicycloplatin at different doses, the cells were collected, fixed, embedded, and sectioned, and the ultrastructural changes of the cells were observed under a transmission electron microscope. . The results showed that the untreated liver cancer cells showed irregular polymorphism, the surface of the cells was rich in microvilli, the organelles in the cytoplasm were abundant, and the glycogen particles were contained in varying amounts. Hepatocarcinoma cells treated with double% platinum showed different degrees of degeneration with the increase of drug concentration. 12 1236478 Pharmacodynamic Z. The double u · cyclopropyl f-diamino complex was also tested several times. The toxicity, the substance ^, and the succinct character of the bicyclic ring (II), which indicate that the bis-U-cyclopropanediamine diamino complex (II) has poor water impurities and higher impurities, but is resistant to __. Pain. Yang 2Cai4 _Specific test characteristics and characteristics of the drug as the active ingredient of the bisdiselamine derivative starting from the present day: 1. Determination of IC50 value a. Take the BGC of the moon cancer -823 cells, cultured in ls% calf serum nutrient solution, inoculated in 96-well culture plates, ix104 cells / well, placed in 3rc, 5% co2 incubator. Dilute the 1% bicyclic transfection solution to the required concentration with RPMi_1640 scale solution containing 15% calf serum, add it to each well of a 96-well plate, each well contains 3 wells, and a 6-well blank control. Incubate for 72 hours and add MTT solution. After 1 hour, DMSO was added to develop the color, and the OD value was measured by a standard instrument. Calculate the kill rate of each concentration of drug, and get the IC50 value according to the coordinate method. Table 2 BGC_823 cell ic50 = 14.80 ± 1.34 pg / ml concentration pg / ml 200 100 50 25 12.5 6.25 3.13 1.56 0.78 kill rate% 83.3 80.3 74.6 60.0 51.0 33.0 21.0 19.0 6.0 IC50 value of human oral cancer welcome cells. Table 3 KB cells IC50 = 13.24 ± 1.33 pg / ml concentration pg / ml 200 100 50 25 12.5 6.25 3.13 1.56 0.78 Killing rate% 93.3 93.0 77.1 66.6 36.7 26.2 28.2 14.8 5.9 13 1236478 2. It has obvious antitumor effect: a ) Forty Kunming mice weighing 21-22 g, half male and half female, were randomly divided into 4 groups of 10 mice each. The right abdomen was subcutaneously inoculated with liver cancer Kb cell suspension 0.2 ml / head. The next day, three groups were given intraperitoneal injection of Shuanghuan 1 from 3Gmg, 2Gmg, and 1Gmg / kg, once a day for 7 consecutive days. The control group was treated with normal saline. Mice were sacrificed the next day after drug withdrawal, weighed and tumor weight was calculated, and tumor suppression rate was calculated. As shown in Table 4, the mice in the drug-treated group gained less weight, but none died, and H22 had a significant antitumor effect. The tumor inhibition rates of 10 mg, 20 mg, and 30 mg / kg were 40%, π%, and 6%, respectively, and the intensity of tumor inhibition was positively correlated with the dose. Table 4 Tumor inhibition rate of H22 for liver cancer

/) Bicyclic uranium measured on sarcoma by the same method as in the above test is shown in Table $. It is shown that double% platinum also has a significant inhibitory effect on Suo, and has a dose-response relationship. Table 5 Grade of tumor inhibition rate of Si80 for sarcoma-^ ^ λ. / 1 mouse front (only) body 1 ~ 1ΓΊ t (g) ~~ ^ Γ ~ tumor weight (g) tumor inhibition rate ------- ^ __ P value 10 10 21.2 29.0 1 · 7 ± 0 · 57 ----- -20mg ^ 10 ~ Ίο ~ --10 10 10 10 21.6 1ΐ · 2 21.5 21.5 ~ 25T ~ 0.49 ± 0.16 0.54 + 0.16 1.03 ± 0 · 30 71.0—68 · Guang—39 · 7 ^ 〇Ό〇Γ ^ 〇όοι_ < 0.01 c) Treatment of human liver cancer BEL_74〇2 cells with 10 pg / ml, 20 μθιη1 bicycloplatinum, and observe the cell dynamics 236478 complain about growth for 8 days, such as As shown in Figure 4, the number of cells in the administration group and the control group was not significantly different within two days. The number of cells in the administration group gradually decreased from the beginning of the next day, while the cancer cells in the control group continued to grow in logarithm. As time increased, the differences between groups increased. It was shown that bicycloplatin had a significant growth inhibitory effect on human liver cancer BEL_74 02 cells, and showed a dose-response relationship. 3. It shows very low toxicity and can selectively kill cancer cells without harming normal cells. A) Take 50 Kunming mice weighing 18_22§, male and female, randomly divided into 5 groups, each group 10 only. Bicycloplatin was administered at 400, 280, 196, 137, and 96 mg / kg, respectively, for one consecutive observation for 10 days, and the general performance and the number of deaths were recorded. Except for high-dose administration, which showed local irritation symptoms, there were no abnormal manifestations, and death usually occurred within 3-7 days. LD5G ± SE = 20.5 ± 1.14 mg / kg was calculated according to the Bliss method. It is significantly higher than the LDsq values of asplatin and carboplatin measured under the same conditions, showing the low toxicity of bicyclic platinum. b) Two rabbits (mg, 3 $ ㈣) of similar age and weight were selected for intravenous injection, and blood samples were taken obliquely, and the initial content in serum samples was measured by atomic absorption spectrometry (AAS method). In Table 6, the concentration of drug in the blood quickly reached a high level after the main shot, exerted its effect, and then decreased with the temple B. After 12 hours, the wave intensity was very low, and it reached 0 in 24 hours. Shuanghuan was able to quickly metabolize and leave the body. Therefore, the toxicity and side effects are very small. Initial content of rabbit Jinqing (pg / ml) Time (hours) Rabbit 1 (3.25kg) Rabbit 2 (3.5kg) 0 34.6 58.0 0.5 20.5 21.8 1 10.4 20.3 2 6.2 10.8 4 2.4 4.2 6 1.9 2.8 ^ 8 1.7 2.3 10-1.6 2.1 Ί 2 1 0.9 2.2 24 〇 0 0 c) Based on the oral acute toxicity test LD5050 mg / kg as a reference, the rats were orally administered 15 1236478 double uranium at LD% / 10. 5% glucose and 0.9% sodium vaporized (ie, sugar saline) were dissolved and administered once daily for 90 days for long-term toxicity experiments. The body weight was measured once a week, and the dose was adjusted according to the weight change. The drug was discontinued after taking the medicine for 12 weeks, and the observation was continued for 2 weeks. The results showed that: (1) There was no death in the rats, and there was no obvious abnormality in diet, activity, and state. This shows that bicycloplatin has no adverse effects on rats. (2) After 2 weeks of discontinuation of the 12-dose administration, the blood collection test showed that the number of red and white blood cells, hemoglobin I, knives, and platelets were in the normal range, and there was no significant difference between the groups (p >005); urea nitrogen ( Β ·), GPT, GOT, there was no significant difference between the administration and the control group (P > 0.05) 〇 (3) After the observation, the rats were sacrificed. The organ coefficient, liver weight of the administration group was slightly higher than that of the blank control group, but still within the normal range. No other organ coefficients were abnormal. (4) Tissue examination of the heart, liver, spleen, lung, kidney, stomach, duodenum, nest, testis, prostate, adrenal gland, thyroid, etc. of the rats. The liver and spleen of the administration group had different degrees of congestion, edema, and other viscera. Is did not see any organic disease caused by the drug. Φ Treat human normal fibroblasts, epidermal cells, lung cancer cells, and melanoma cells with bicycloplatin. The experimental results are shown in ® 5. In the-fixed dose range, normal f fiber cells and epidermal cells are not injured, while lung cancer cells and melanin are not damaged. The tumor cells will be killed. For example, when the concentration of bicycloplatin was 0.00347 mg / i, lung cancer cells and melanocytes were killed by 50% and 8G%, respectively, showing that bicycloplatin can selectively kill cancer cells without harming normal cells. . E) After the human liver cancer BEL-7402 cells were treated with bicycloplatin, the ultrastructural changes of the cells were observed under a transmission electron microscope. Non-medicated liver cancer cells are irregularly polygonal and have rich microvilli on the surface: the cytoplasm is full of organelles and contains varying amounts of glycogen particles (Figures 6, 7). Hepatocellular carcinoma cells treated with bicyclic platinum showed different degrees of degenerative changes, a large number of vacuoles appeared in the cytoplasm (Figure 8), and lipid droplets were large (囷 9) until necrotic changes, nuclear shrinkage, nuclear fragmentation, and nuclear Dissolved, the organelles completely disintegrated (dead after 5 days of death. 5 Typical anti-cancer drugs Shuanghuan platinum oral capsules and injections clinical typical cases: 16 1236478 ι · Shuanghuan and auxiliary materials composed of oral capsules, patients with oral capsules containing 20-30 mg of bicyclic platinum. Healing Case: Zhu, male, 64 years old, bladder cancer. After finding that the hematuria laser treatment department has recurred three times, he started taking Shuanghuan. The original 3 pieces of 2x2cm2 lumps completely disappeared. No abnormalities were found in both kidneys and bladder. Wang's, female, 65 Aged, liver cancer, advanced, critically ill. After taking bicycloplatin and intravenous injection of 30 mg A butin, the left and right tumors of the liver were 4.7 x 3.1 cm, and after 4 to 5 days of administration, the tumor shrank to 2.3 x 1.7 cm. The tumor disappeared and the disease Healed, life is normal. 2 · Bicyclic uranium water injection, 5ml ampoule, bicyclic platinum content 1%, once every other day, 2 each time. Bicyclic platinum water solution is dissolved in 100ml 5% glucose 〇9% sodium chloride injection After dripping, vaporize with 100ml The injection solution is dripped, 6 times (12 ampoules) as a course of treatment. Two weeks later, a second course of treatment is performed. After 3 consecutive courses of treatment, a comprehensive examination is performed. Ma, male, 83 years old, with advanced recurrence of liver cancer Lungs. Since then, I started taking Shuanghuan Platinum Capsules 4 times a day, 4-6 capsules each time. After 2 months of taking, the metastatic lung cancer tumors went from 2x2cm to disappeared. The bone metastatic cancer tumors shrank from 18x14cm to 14xllcm. Front smooth and slightly shrunk

㈣t-like mental state and health, weight ㈣Kg at 6lKg, little change in contrast before and after blood treatment, the treatment effect is good. Xu, male, 59 years old, with lymphatic metastasis. When she relapsed, she had a low score of nasopharyngeal cancer, which recurred years after the needle was placed, and the nasopharyngeal cancer was advanced.

Common reactions to other anticancer drugs such as heart and vomiting. Hepato-pulmonary metastasis, Yang, male, 45 years old, melanoma, 17 1236478 after injection of 36 needles of bicyclic platinum injection The tumor is necrotic. . Li Gan, male, and 9-year-old 'primary liver cancer' tumor volume 3.3 > < 3.3cm bile duct metastasis portal vein with double ring treatment for three courses of% branch, the first course of the left liver lobe tumor volume shrinks ", '3X3'3Cm' In the second course of treatment, the tumor of the left hepatic lobe basically disappeared, and the right bile duct only expanded by 0.8 cm, and portal cancer was eliminated. Chu Yi J Paper Continued The course of treatment is stable, the patient is mentally sad and appetite is good, little change before and after blood treatment. Periodic fever for about 10 hours and sometimes pain in the ward. Meng, a 'male', was 48 years old. He was diagnosed with advanced lung adenocarcinoma as Stage I V by the 301 hospital. He was given bicycloplatin and needles at the same time. Shuanghuan capsules were all taken orally. All symptoms such as coughing, wheezing, and phlegm had disappeared. ^ A foreign patient 'male, 60 years old, Russian and Chinese Academy of Medical Sciences Cancer Hospital and Air Force General Hospital did #hepatic metastasis of colon cancer and yellow gallbladder appeared. Doctors in Russia suggested to go to China for treatment after surgery. 4 shots recommended 36 injections of double injections and 36 capsules of Shuang Na Na capsule at the same time, the liver metastasis disappeared, and has now been basically cured. From the above large-scale scientific experiments, from ex vivo cell experiments, clinical (both oral and injection injection routes),, oral fruit and heart, long-term toxicity, and experimental conclusions on normal cells have proved that the double Dicarboxamidine ... starter derivatives (especially bicyclic starter and bis-, cyclopropanyl diammonium diammonate starter) are a generation of safe and effective anti-facial drugs, bringing new hope for humans to overcome cancer. The following briefly describes the drawings: Figure 1 is the infrared spectrum of bicyclic platinum; Figure 2A is the 1H_NMR spectrum of bicyclic platinum; Figures 2B and 2C are the negative ion ESI_Ms (electrospray mass spectrometry) of bicyclic platinum; Figure 3 is X-ray quaternary diffraction unit cell stacking diagram of Shuanghuanming; Figure 4 is the growth curve of human liver cancer BEL-7402 cell with bicycloplatin; Figure 5 is the curve of selective killing of cancer cells with bicyclic molybdenum; Figure 6 is a blank control BEL-7402 cells (ΕΜχΙΟΟΟΟ); Figure 7 is a blank control of BEL-7402 cells (ΕΜχ6000); 1236478 Figure 8 is a vacuolar transformation of BEL-7402 cells (ΕΜχ12000); Figure 9 is a fatty degeneration of BEL-7402 cells (ΕΜχ 12000) Figure 10 shows BEL-7402 nuclear fragmentation (EMX6000); Figure 11 shows BEL-7402 nuclear fragmentation (EMX8000). Explanation and assignment of the main peaks in Figure 1: Group 3291cm4 is the stretching vibration peak of Pt—NH3; 2539-2992cm · 1 is a peak that rises slowly, instead of the -COO- group of carboxylate;

1716. The strong peak of HiHcm4 is in the carboxylic acid group in the bicyclic platinum < 1_, I56-1. The strong peak is c = 〇 stretching vibration in _C00_Pt in the molecule / the strong absorption peak of 1400cm · 1 is _c = 〇 The variable-angle vibration absorption peaks;, peaks in the fingerprint region of 697 ~ 1317cmi, not explained one by one. Ammonia platinum (II) (referred to as bicyclic platinum) The above infrared spectrum data is a fingerprint characterization of bisU-cyclobutyldicarboxydi. The 1H-NjyfR analysis of the bicyclic platinum in Fig. 2A is as follows: δΡΡιη Peak shape JHz Η Number of proton groups explained 1.64 5 heavy peaks 7.5 2 Structure (II) 2 magnetically equivalent Η 3 in CH2 of the right quaternary ring 1.84 5 Heavy peak 7.5 2 structure (II) 2 magnetic equivalents of 3 CH2 in the left four-membered ring 36 2.36 3 heavy peak 7.5 4 structure (II) 4 magnetic equivalents of 2 and 4 digits in the right four-membered ring Η 2.49 multiplet-未 undeuterated Η in solvent DMSO 2.65 3 heavy peaks 7.5 4 structure (II) left four-membered ring 2, 4 digits 2 magnetic equivalents of 4 CH 19 ^ 236478 3.28 singlet one 4.06 Singlet — 6 12.61 Singlet 2 * From the above analysis data and bicyclic platinum (π) Bicyclic platinum molecular weight: 515.36. Trace Η20 in solvent DMSO Structure (II) Two νη3 6 Pt on Pt Magnetic equivalence 两个 Two -COOH magnetic equivalence Η in Structure (II) The structure shown is completely consistent. Figure 2B clearly shows the para-excimer ion peak wall 514. The benefits represented by all the peaks in Figure 2C are reasonable in terms of the structure of the financial compound. In FIG. 4, “.” Indicates a control group, “◊” indicates 1 (Wml, “χ,” indicates Zheng Kai. In FIG. 5, “.” Indicates a normal human fibroblast, and “·” indicates a normal human epidermal cell. The human melanoma cells, "□", represent human lung cancer cells. The present invention can be further illustrated by the following examples of preparing the derivative of formula (I) and its pharmaceutical composition: Example 1: Double Preparation of 1,1-cyclobutanedicarboxylic acid diammineplatinum compound Add 3.54 g (10.0 mmol) of carbohydrate to the reaction flask, add 200 mi of pure water, and stir to avoid dissolution until it is completely dissolved. 16.2g (10.0mmol) of citronella acid, stir, dissolve, rotate to dryness, add 50ml of ethanol, stir for 2 hours, filter, wash with 10mlx3 ethanol, recrystallize with distilled water, evacuate, and dry naturally. Example 2: Preparation of bis-1,1-cyclopropanedicarboxylic acid diammine platinum compound In a reaction flask, Ag6.5S (40.8 g, 20.8 mmol) was added, and 100 ml of purified water was added to the reaction flask. Completely dissolve, add 26.2g (20.7mmol) of Cis- (NH3) 2PtCl in batches with stirring, and stir the reaction for 5 hours in a water bath below 40 ° C. Filter while hot, wash with 10 ml × 3 warm water, transfer the filtrate to the reaction flask, add 35% (H20 220 ml (232.6 mmol)) in portions, stir the reaction at room temperature for 4 hours, and warm to 60 ° C for 2 hours. Then add 12.0 g (25.0 mmol) of barium cyclomalonate, search and stir at 60 ° C for 8 hours, filter while hot, wash with 10 ml × 3 warm water, transfer the filtrate to a reaction bottle, and spin-dry 1236478 to dry, Add 50ml of ethanol, stir for 2 hours, filter, wash with 0mlx3 ethanol, recrystallize distilled water, evacuate, and dry naturally to obtain the title compound. Decompose at 220 ° C. Example 3: Bis U-cyclobutanedicarboxylic acid diamine complex Preparation of platinum compound 105g (61.8mmol) AgNO3, add 100ml of pure water, stir in a reaction bottle until dissolved, add 9.3g (31.1mmol) of diaminoplatinum in portions, and stir the reaction below 40 ° C. 5 Hours. Filter while hot, wash with 10mlx3 warm water, transfer the filtrate to the reaction flask, add 35% H2O2 30ml (349.0mmol) in batches (15ml + 15mlx3), stir the reaction for 5 hours at room temperature, warm to 6 (rC reaction 2 hours. Then add 12.0g (69.0mmd) of succinic acid, 6 (TC stirring reaction for 8 hours, filter while hot, wash with 10mlx3 warm water, and transfer the filtrate In a reaction flask, rotary evaporation to dryness, 50 ml of ethanol was added, and the mixture was stirred for 2 hours, filtered, washed with 10 ml × 3 ethanol, distilled water was recrystallized, evacuated, and naturally dried to obtain the title compound. Example 4: Double 1,1-ring The same procedure as in Example 3 was used to prepare butanedicarboxylic acid diammine platinum compound, and 15.0 g of diiodine diammine platinum was used instead of 9.3 g of digas diammine platinum to obtain the title compound. Example 5: Preparation of bis 1,1-cyclopentanedicarboxylic acid diammine platinum compound iO.Sgai.Smmo ^ AgNO3, add 100 ml of pure water, stir in a reaction ampoule until dissolved, and add two gas two Aminoplatinum 9.3g (3Ummol), the reaction was stirred for 5 hours below 40 ° C. Transition while hot, wash with 10mlx3 warm water, transfer the filtrate to the reaction flask, add 35% H202 30ml (349.0mmol) in batches (15ml + 15mlx3), stir the reaction for 5 hours at room temperature, and heat to 60 ° C for 2 hours. Then add 15.6 g (53.0 mmol) of barium glutarate (the cycloglutarate is a known substance and it can be prepared by known methods) '60 C Stir; stir for 8 hours, filter while hot, wash with 10 ml x 3 warm water The liquid was transferred into a reaction flask, rotovaped to dryness, added with 50m of ethanol and stirred for 2 hours, filtered, washed with 10mlx3 ethanol, recrystallized from distilled water, evacuated, and naturally dried to obtain the title compound, and its melting point was 197. 〇 ~ 199〇C 〇 Example 6 · Preparation of bis 1,1-cyclohexane dicarboxylic acid diammine platinum compound 6.7 g (39.4 mmol) AgN 03 'Add 100 ml of pure water, Stir until dissolved, add in batches. 6.2g (19.6mmol) '40 C is allowed to react for 5 hours. Tear while hot, wash with 21 1236478 1〇1111 > < 3 warm water, transfer the filtrate to a reaction flask, add 350 / 〇112022〇1111 (232.6111111〇1) in batches, stir the reaction at room temperature for 5 hours, and raise the temperature to 60 ° C reaction for 2 hours. Then 3.7 g (21.5 mmol) of cycloadipate was added (the cycloadipate is a known substance which can be prepared by known methods). 45. (: With stirring, slowly add 1 ml (10.3 mmol) of hydrazine hydrate. Dropping is completed within 2 hours, the reaction is continued for 6 hours, rotary evaporation to dryness, and vacuum dehydration and drying for 5 hours. Add 50 ml of methanol and stir for 2 hours, filter, lmix3 Wash with methanol, recrystallize with distilled water, and vacuum dry to obtain the title compound, and its melting point is 212 ° C ~ 214 ° C. Example 7: Preparation of bis 1,1-cyclobutanedicarboxylic acid diaminoplatinum and Refined 12.4g of digas diaminoplatin was suspended in 800ml of pure water, and a stoichiometric 1N AgNCb solution was added under stirring and protected from light, and reacted at 40 ° C for 4 hours. The refrigerator was left to filter. The precipitate was removed by filtration (recovered for future use). Stoichiometric cyclobutanedicarboxylic acid was added to the filtrate, and the reaction was stirred for 16 hours at 6 ° C., and evaporated under reduced pressure until microcrystals appeared. The refrigerator was left for 48 hours, filtered, and the crystals were collected to obtain a crude product. Refined and purified to obtain a refined product yield of 90%, melting point 182 ° C ~ 184. (: Preparation Example 1: 50 g of bicyclic injection is first dissolved in 5000 ml of pure water, 7.5 g of medical charcoal is adsorbed and filtered, and a 2um filter membrane is used. Filter twice, dispense into 1000 5ml ampoules, and seal by burning Injection Example 2: Preparation of oral preparation: 24 g of duraplatin, i600 g of medical starch and 120 g of vitamin c, mixed and crushed in a blender, if necessary, divided into 5,000 oral capsules, An oral preparation was obtained. The formula ⑼ shuanghuan prepared in the above implementation of m, 3, 4, and 7 is colorless needle-like crystals, and the micelles are capped into white microcrystalline powders. Measured by melting point, solubility, and pH, carbon, gas, and nitrogen Analysis of trace elements of ICP_EAS by trace element analysis, differential thermal and thermal weight loss analysis of infrared spectrum, ultraviolet spectrum, laser Raman spectrum, thin-layer chromatography, lH,%, 195pt_ total series, χ-quaternary complement, etc. The chemical method has fully tested the composition, structure, and purity of the formula (Cong_). The double-i small ring sintered two-ship diamino complex Lina single crystal structure of formula (II) has been confirmed by Gu Wei. For the domain-like compound, the infrared spectrum, 22 1236478 iH-NMR, MS and X-quaternary diffraction pattern are shown in the attached drawings 1 to 3 and the following Table 1. Table 1 X-ray quaternary diffraction measurement results Single crystal size (mm) 0.2x0.2x0.3 Crystalline monoclinic space group P21 / C Cell parameters a = 5.679 (2) A b = 12.107 (2 ) A c = 23.430 (5) A β = 95.25⑵0 Volume 1604.3⑺ A3 Density 2.123Mg / m3 Absorption Coefficient 8.876mm " 1 Number of Unit Cells 4 The new formula (I) bisaminodiamine diammine platinum Derivatives have an unexpected effect in the treatment of cancer. Compared with the existing platinum-based anticancer drugs, the formula ① diaminodiamine platinum derivative of the present invention is characterized by: · The compound is formed by four hydrogen bonds in the molecule The cage-like molecular structure locks platinum atoms in the cage, which makes the compound have a certain stability, weakens the attack of water molecules and ensures the stability in water. It well overcomes the complexity of the compound and the lethal effect of Shui Lin Zhi, and retains the significant advantages of its medicinal effect, greatly improves the water solubility, and is stable in water. It can be used subcutaneously in the abdominal cavity, veins, and pi. Dosing. It not only has higher resistance, but also greatly reduces /, and also overcomes the instability of its water towel. Its significant features are as follows, taking the bicyclic platinum compound of formula (II) as an example: 1. High anticancer activity, such as IC50 stuck to breast cancer cells in white 9.30μ8 / πι1 doubled doubled 2.80pg / ml to liver cancer cell BL7402 IQo carboplatin 8.45μΕ / ηι1 Bicyclic Molybdenum 1.30pg / ml H Yueguang pairs / birth cancer, head and neck cancer, nasopharyngeal cancer, breast cancer, lung cancer, liver cancer, pancreatic cancer, 23 1236478 gastric cancer, intestinal cancer, lymphatic cancer, etc. have significant effects . 3, low toxicity, LD50 intraperitoneal injection of 13mg / Kg Kagu 130mg / Kg shuanghuan 283mg / Kg oral shuanghuan 500mg / Kg (cisuranium and carboplatin are not effective orally) The clinical test toxicity is 0 degrees to i degrees ( Classified according to the fifth degree), there were no adverse reactions in clinical patients. 4. Good water solubility, good stability, water solubility (g / 1〇ml) cisplatin 〇2 carboplatin 1.5 bicyclic platinum 4.0 cisplatin and cardosol dissolve in water-day after day, while the bicyclic ring of formula ⑼ is homozygous The product can be stored for a long time without failure. 5. It is effective when taken orally, and the derivatives of Shuang Er Jian Di Ammonia are not only effective but also snail. For example, a capsule made of the bicyclomine compound of formula (II) as the main agent and supplemented with a pharmaceutically acceptable excipient has undergone a long-term and large-scale clinical oral trial and has a significant effect.

24 1236478 [Schematic description]

Claims (1)

1236478 The scope of patent application: 1 ·-a kind of bis-divisco-diammine derivative, which is characterized in that it has the formula 1: R1 > < R2 / 〇 Η-〇c < VH C-〇X meaning: Η &Η; Η Η, Η〆, Η 'Η- \ / R1 C, R2 where A and R2 are connected to each other and connected to them. Together, they form a saturated carbocyclic ring containing carbon atoms. 2. The bi-second-spectrum diammonium hydrazine organism according to item i in the scope of the patent application, characterized in that it is a bi-u-cyclopropane or di-un-diamine compound. 3. The shuangdijian diamino complex as described in item 2 of the scope of the patent application, characterized in that: / is a bis 1,]-cyclobutadiene diamino complex compound of the following formula (II) : 'The described seed is used for the method for preparing the bis-dianylidene complex, which is characterized in that the organism has H 生物:, 〇 ----- Η——〇 R' R2: xr > ^ c—ο 〇 ~ \ / Η ,,, Η〆'Η -Ή—— \ / R1 C CT and R2 formulas. R and r2 are connected to each other and together with the carbon atoms to which they are connected form a 3-6 1236478-membered saturated carbocyclic ring containing carbon atoms. The method is: carboplatin or carboplatin Reacting with a dicarboxylic acid ligand derivative of the following formula (III) R1 〇II C—OH C (ΠΙ) r2 / C—OH IIο where R1 and R2 are defined as described in formula (I) above to give formula ( I) a bisaminodiaminoplatinum derivative; or 2) a dihalodiaminoplatinum of the formula (NH3) 2PtX2 where: X is C1 or I and silver nitrate or silver sulfate is reacted in an aqueous solution to produce a formula: (NH3) 2Pt (H20) 2 (N03) 2 hydrated diammine nitrate or hydrated diamine starting sulfate (NH3) 2Pt (H20) 2 (S04); the hydrated diammonium platinum nitrate or Sulfate reacts with a dicarboxylic acid ligand derivative of the following formula (III) or its sodium or barium salt R1 0 II C—OH C (ΠΙ) r2 / C—OH IIο where: R1 and R2 are as defined above The bis dicarboxylic acid diamino complex platinum derivative of formula (I) is described in formula (I). • An antitumor pharmaceutical composition, characterized in that the composition contains at least one bis-dicarboxylic acid diaminoplatinum derivative of the formula α) as described in claim 1 in an amount of 0.001-55% by weight at 3 1236478 degrees. Physically acceptable carrier. Le Wu 6 · The pharmaceutical composition according to item 5 of the scope of patent application, characterized in that the bis-hybrid acid diammine platinum derivative of formula (I) is bis 1,1-cyclopropane dicarboxylate Acid diammine platinum or double! L-cyclobutane diamino complex platinum compound. The pharmaceutical composition according to item 6 of Shuangyi slow moon r Zhuwei, characterized in that the formula ⑴ 8. If the application domain is a bis-U_cyclobutadiene_diammine compound. Seeking Yuanyuan No. 5 to No. Pharmaceutical composition The pharmaceutical composition according to any one of the items, characterized in that. Administration in the form of injection capsules. For example, the application of the patent application in the Fanyuan patent, the elk drug in μ tumor drugs, the bisaminodiaminoplatinum derivative of formula (I) in the preparation technology