CN1187047C - Application of difenitriazole in preparation of medicine for treating malignant tumour - Google Patents

Abstract

The present invention discloses the application of difenitriazole to the preparation of a medicament for treating malignant tumors. Difenitriazole is a triazole compound, and the chemical name is 3-(2-ethylphenyl)-5-(3-methoxyphenyl)-1H-1, 2, 4-triazole. The difenitriazole has stronger out-of-body inhibiting action on various tumour cells, such as a human ovarian cancer HO-8910 cell, a mouse lymphoid tumour P388 cell, a human oral cavity epidermoid carcinoma KB cell, an ileocecum adenocarcinoma HCT-116 cell, a chronic marrow tropocollagen leukemia K562 cell, a human lung cancer A549 cell, a human nasopharyngeal carcinoma CNE cell, a human cervical carcinoma Hela cell, a human malignant embryo rhabdomyoma RD cell, an original medullary cell leukemia HL-60 cell, a prostatic cancer PC-3 cell, a human liver cancer Bel-7402 cell, etc., and can be applied to the preparation of various malignant tumor drugs.

Description

Application of defetriazole in preparation of medicament for treating malignant tumor diseases

Technical field

The invention relates to a new application of defetriazole in the field of pharmacy.

Background technique

Difetriazole was first reported by Italian Galliani in 1978, and its synthetic route was found to have antifertility activity. Defetriazole has been reported and used as a method for terminating early pregnancy. The drug has no official name in Italy, and Galliani et al. used the code name DL111-IT. After searching, there were 4 foreign patents, namely West German patents 2819372 (1978), 2943326 (1980); US patents 4379155 (1978), 4370336 (1983). Published the preparation method of 3,5-disubstituted 1H-1,2,4 triazole derivatives, a total of more than 40 derivatives (including DL111-IT), and proved to have antifertility and sedative effects on animals.

Diphenytriazol (DPTZ for short), chemical name 3-(2-ethylphenyl)-5-(3-methoxyphenyl)-1H-1,2,4-triazole, English chemistry The name is 3-(2-ethylphenyl)-5-(3-methoxyphenyl)-1H-1,2,4 triazol. Its chemical structural formula is:

Molecular formula: C ₁₇ H ₁₇ N ₃ O Molecular weight: 279.34

Physical and chemical properties of this product: white crystalline powder. Odorless, tasteless, easily soluble in absolute ethanol and chloroform, slightly soluble in vegetable oil, hardly soluble in water. Melting point: 76-81°C.

Contents of Invention

The object of the present invention is to provide a new application of defetriazole in treating malignant tumor diseases, that is, a new application in pharmacy.

Specifically, the present invention relates to defetriazole as a preparation for treating various malignant tumor diseases such as ovarian cancer, cervical cancer, lymphoma, leukemia, lung cancer, liver cancer, colorectal cancer, prostate cancer, nasopharyngeal cancer, oral cancer, and malignant rhabdomyoma. application in medicine.

Preparation of Defetriazole Injection

Take 1000ml of vegetable oil for injection (tea oil, soybean oil, etc.), dry heat sterilization at 150°C for 1 hour, let it cool to 50°C, take out part of the oil and add 1g of difetriazole, stir to dissolve, and finally add solvent to the full amount . Filter it with a dry No. 3 vertical melting funnel, fill it in a 1ml dry sterilized ampoule, and sterilize it with circulating steam at 100°C for 30 minutes to prepare an injection solution containing 1 mg of difetriazole in 1 ml of each ampoule.

Preparation of Difetriazole Nano-emulsion Pills

Dissolve defetriazole in non-aqueous liquid or semi-dry solid compound excipients to prepare soft capsules, each pill contains 0.5mg defetriazole

Once this composite excipient is mixed with water or gastrointestinal fluids, it will rapidly and spontaneously form a nano-sized oil-in-water emulsion, and the drug is dissolved in these extremely small oil droplets and rapidly dispersed to the absorption site in the gastrointestinal tract . During the entire absorption process, the drug remains in a molecular state, and these tiny drug-containing oil droplets can increase its adsorption on the intestinal epithelial cells, prolong the absorption time, increase the transport of the drug through the lymphatic system and pass through the Payer's area of the intestinal tract. Phagocytosis into the body's circulation avoids first-pass elimination. Improved bioavailability and increased drug targeting.

Preparation of Difetriazole Microsphere Injection

Biocompatible and biodegradable polymers are prepared into desfetriazole injection microspheres, and each injection contains 2 mg of desfestriazole.

After intramuscular injection, the polymer will slowly release and absorb the drug at the administration site, prolonging the action time. This polymer is hydrolyzed into harmless degradation products (water and carbon dioxide) after drug release. long-acting non-triazole

Intramuscular injection of microsphere injection once can maintain the drug effect for one month. It is used for the prevention of metastasis and recurrence of cancer cells after operation of malignant tumors.

In order to better understand the essence of the present invention, the following pharmacological and toxicological tests and results of defetriazole will illustrate its new use in the field of pharmacy.

1. In vitro inhibitory effect of difetriazole on various tumor cells

Take a bottle of human ovarian cancer HO-8910 cells that have been cultured for 3 to 4 days, add an appropriate amount of trypsin to make the adherent cells fall off, make a cell suspension with RPMI1640 nutrient solution containing 10% fetal bovine serum, and use complete culture medium after counting Dilute to a cell suspension with a concentration of about 4× ¹⁰⁴ /ml, take a 24-well plate, add 1ml of cell suspension to each well, put it in the incubator for 48 hours, add 10μl of the test substance, so that the final concentrations are: Defetriazole 40μg/ml, 20μg/ml, 10μg/ml, 5μg/ml, 2.5μg/ml, 4 wells for each concentration, cultured for 48 hours after adding the drug, added 5mg/ml MTT60μl to each well, incubated for 4 hours Aspirate the supernatant, add 0.5ml DMSO and shake well, measure the OD value of each small well at 550nm with a microplate reader, the formula for calculating the cell survival rate is:

Cell viability%=(OD value of drug-added cells/OD value of control cells)×100%.

IC ₅₀ was calculated by Bliss method. repeat three times.

Mouse lymphoid tumor P388 cells, human oral epidermoid carcinoma KB cells, ileocecal adenocarcinoma HCT-116 cells, chronic myelogenous leukemia K562 cells, human nasopharyngeal carcinoma CNE cells, human cervical cancer Hela cells, human malignant embryonic striated muscle The in vitro inhibition test method of tumor RD cells, human lung cancer A549 cells, myeloid leukemia HL-60 cells, prostate cancer PC-3 cells, and human liver cancer Bel-7402 cells is the same as above.

The results showed that after co-incubating cells such as HO-8910, P388, KB, HCT-116, K562, CNE, RD, A549, Hela, HL-60, PC-3, Bel-7402 for 48 hours, difetriazole had Show strong inhibitory effect on cell proliferation. After 40 μg/ml of difetriazole acted on the above tumor cells for 48 hours, the inhibition rates were 94.1%, 98.6%, 87.1%, 99.7%, 96.5%, 65.5%, 98.0%, 68.7%, 92.3%, 90.0% , 93.9%, 100.0%, and their IC ₅₀ and 95% confidence limits were 15.1 (12.5-18.4), 4.1 (1.3-13.5), 10.7 (7.4-15.6), 6.2 (1.8-21.6), 9.9 (5.3- 18.4), 17.3 (10.5-28.5), 12.6 (8.0-20.2), 17.4 (13.0-23.4), 9.9 (4.8-19.6), 14.1 (7.4-26.7), 19.7 (8.6-40.5), 5.6 (1.4-15.7 ) μg/ml. See Table 1.

Table 1. The in vitro inhibitory effect of difetriazole on various tumor cells for 48 hours

Tumor Defetriazole (μg/ml) IC ₅₀ and 95% confidence limit

Cells 2.5 5.0 10 20 40 (μg/ml)

HO-8910 0.7 6.1 28.9 57.6 94.1 15.1(12.5-18.4)

CNE 11.3 23.4 47.7 51.7 65.5 17.3(10.5-28.5)

A549 18.3 22.8 40.1 50.4 68.7 17.4(13.0-23.4)

P388 42.1 60.0 68.3 78.2 98.6 4.1(1.3-13.5)

K562 14.2 17.6 32.2 71.0 96.5 9.9(5.3-18.4)

Hela 13.7 21.9 39.5 72.6 92.3 9.9 (4.8-19.6)

HL-60 - 2.4 26.9 85.5 90.0 14.1(7.4-26.7)

PC-3 - - - 6.6 50.6 93.9 19.7(8.6-40.5)

Bel-7402 31.2 41.3 62.7 89.2 100.0 5.6(1.4-15.7)

KB 15.7 28.2 49.2 57.7 87.1 10.7(7.4-15.6)

RD 1.6 11.3 27.7 54.7 98.0 12.6(8.0-20.0)

2. The time-effect relationship of defetriazole's inhibitory effect on human ovarian cancer HO-8910 cells in vitro

Take a bottle of HO-8910 cells that have been cultured for 4 to 5 days, add an appropriate amount of trypsin to make the adherent cells fall off, make a cell suspension with RPMI1640 nutrient solution containing 10% fetal bovine serum, and dilute it with complete culture medium to obtain a concentration after counting. About 4× ¹⁰⁴ cells/ml of cell suspension, take 24-well plate, add 1ml of cell suspension to each well, put it in the incubator for 48 hours, add 10μl of the test substance, so that the final concentrations are: Azole 40 μg/ml, 20 μg/ml, 10 μg/ml, 5 μg/ml, 2.5 μg/ml. 4 wells for each concentration. After adding the drug, culture for 24 hours, 48 hours, 72 hours, and 96 hours, add 5mg/ml MTT60μl to each well, incubate for 4 hours, suck off the supernatant, add 0.5ml DMSO and shake well, suck out 0.1ml from each well into a 96-well plate, Use a microplate reader to measure the OD value of each small well at 550nm, and the formula for calculating the cell viability is:

Cell viability%=(OD value of drug-added cells/OD value of control cells)×100%.

IC ₅₀ was calculated by Bliss method. Repeat each action time 3 times.

The results showed that desfetriazole showed a strong anti-proliferation effect on human ovarian cancer HO-8910 cells for 24, 48, 72, and 96 hours. The inhibitory rates of defetriazole 40 μg/ml on HO-8910 cells for 24, 48, 72, and 96 hours were 79.2%, 94.1%, 94.2%, and 93.6%, respectively, and the IC ₅₀ and 95% confidence limits of the above time points They were 23.9 (13.5-42.3), 15.1 (12.5-18.4), 12.4 (9.7-15.7), and 9.1 (6.3-13.2) μg/ml, respectively. See Table 2.

Table 2. Time-effect relationship of in vitro inhibitory effect of difetriazole on human ovarian cancer HO-8910 cells

Action time DL111-IT(μg/ml) IC ₅₀ and 95% confidence limit

(h) 2.5 5.0 10 20 40 (μg/ml)

24 0.1±0.1 4.3±5.6 6.2±6.9 36.3±7.7 79.2±11.9 23.9(13.5-42.3)

48 0.7±0.6 6.1±0.9 28.9±3.2 57.6±23.4 94.1±1.5 15.1(12.5-18.4)

72 2.6±2.9 5.8±3.3 40.7±13.1 80.1±11.3 94.2±2.0 12.4(9.7-15.7)

96 7.4±8.4 14.4±11.2 59.1±13.6 90.4±6.6 93.6±5.4 9.1(6.3-13.2)

3. Effect of defetriazole on the growth curve of human ovarian cancer HO-8910 cells

Take HO-8910 cells in the logarithmic growth phase and make 3× ¹⁰⁴ cells/ml cell suspension with complete culture medium, take 24 25ml culture bottles, add 5ml cell suspension to each bottle, divide into 4 groups, 6 bottles in each group , respectively (1) solvent control group, (2), (3) administration group: the final concentrations of difetriazole were 1.5 μg/ml and 6.0 μg/ml respectively, (4) positive control group: cisplatin 1.0 μg /ml. Put them in an incubator, cultivate them for 1, 2, 3, 4, 5, and 6 days, and then take 1 bottle for each group, digest with trypsin to make the adherent cells fall off, use 0.7ml of complete culture medium to make a cell suspension, and then Add 0.3ml of 0.4% trypan blue solution, count, plot the logarithm of the number of living cells rejected per ml and time, read the growth saturation density (N _s ) from the graph, and calculate the killing rate of cells according to the following formula and cell doubling.

Cell killing rate of difetriazole (%)=(N ₀ -N ₀ ′)/N ₀ ×100

Effect of difetriazole on cell doubling time (TD): TD=0.301t/(logN _t -logN ₀ ) Cells in each group were stained with trypan blue after cultured for 1, 2, 3, 4, 5, and 6 days, and counted , the logarithm of the number of living cells rejected per milliliter and time was used as a growth curve, and it can be drawn from the figure that the killing rates of difetriazole 1.5pg/ml and 6μg/ml to HO-8910 cells were 14.1 % and 23.4%, the saturation density of cell growth (N _s ) was significantly decreased (p<0.05), and defetriazole 6 μg/ml significantly prolonged the cell doubling time (TD) (p<0.05), see Table 3. In the table: ^* P<0.05, vs DMSO group, t test.

Table 3. Effect of difetriazole on the growth of human ovarian cancer HO-8910 cells

Drug Concentration Cell Doubling Time (TD) Killing Percentage Cell Growth Saturation Density (N _s )

(μg/ml) (h) (%) (×10 ⁵ cells/ml)

DMSO80μl 36.8 ±2.4 - 3.70±0.40

Difetriazole 1.5 40.7±1.8 14.1±3.0 2.64±0.31 ^*

Difetriazole 6.0 44.8±3.1 ^* 23.4±4.4 2.21±0.17 ^*

4. Therapeutic effect of defetriazole on mouse H22 liver cancer and mouse S180 sarcoma

The mouse sarcoma S ₁₈₀ and mouse liver cancer H22 tumor cell suspensions were inoculated in the right axilla of NIH mice respectively, and 60 mice were inoculated for each tumor, half male and half male. 60 mice were randomly divided into five groups, respectively positive control cyclophosphamide 50mg/kg group, defetriazole high-dose group (50mg/kg), middle-dose group (25mg/kg), low-dose group (12.5mg/kg). kg) and the solvent control group (tea oil 10ml/kg). Continuous administration for 9 days, on the 10th

The mice were dissected, the tumor tissues were taken out and weighed, and the tumor inhibition rate was calculated.

The results show that defetriazole has a good therapeutic effect on mouse S180 sarcoma and mouse H22 liver cancer, and the tumor inhibition rate of defetriazole 12.5, 25.0, 50.0 mg/kg on mouse S180 sarcoma reached 33.55% respectively. %, 50.00% and 55.92%, and the tumor inhibitory rates against mouse H22 liver cancer reached 24.50%, 52.00% and 56.60%, respectively. See Tables 4 and 5. In the table: ^** P<0.01, vs camellia oil group, t test.

Table 4. Therapeutic effect of difetriazole on mouse S180 sarcoma

Group Dose Dose Administration Number of Animals Tumor Weight Tumor Inhibition Rate

(mg/kg) Route (only) (g) (%)

Camellia oil control 10ml/kg im 12 1.52±0.57 -

Defetriazole 50.0 im 12 0.67±0.35 ^** 55.92

25.0 im 12 0.76±0.40 ^** 50.00

                                                                                                

Cyclophosphamide 50.0 im 12 0.83±0.25 ^** 45.39

Table 5. Therapeutic effect of difetriazole on mouse H22 liver cancer

Group Dose Administration Administration Number of Animals Tumor Weight Tumor Inhibition Rate

(mg/kg) Route (only) (g) (%)

Camellia oil control 10ml/kg im 12 2.00±0.49 -

Defetriazole 50.0 im 12 0.87±0.33 ^** 56.60

25.0 im 12 0.96±0.46 ^** 52.00

                                                  

Cyclophosphamide 50.0 im 12 1.06±0.50 ^** 47.00

5. Therapeutic effect of defetriazole on human ovarian cancer HO-8910 xenografted tumor in nude mice

Vigorously growing human ovarian cancer HO-8910 cells were digested to prepare a cell suspension with a concentration of 10 ⁷ cells/ml, and 0.2ml of the cell suspension was inoculated subcutaneously in the right armpit of nude mice with a syringe under sterile conditions. 55 nude mice were randomly divided into 4 groups, respectively 50.0, 25.0, 12.5mg/kg/day of defetriazole, 50mg/kg/day of positive control cyclophosphamide, and 10ml/kg/day of solvent control tea oil. Both groups were administered by intramuscular injection (im), administered twice a week for a total of 4 weeks, and the diameter of the tumor was measured twice a week during the period of administration.

Product (TV) is calculated as follows: $\mathrm{TV}=\frac{1}{2}a{b}^2,$ Where a and b represent length and width, respectively. On the 28th day, the tumors were dissected, and the tumors were weighed, and the tumor inhibition percentages of each administration group were calculated.

The results showed that difetriazole 12.5, 25.0, and 50.0 mg/kg had different degrees of inhibitory effects on human ovarian cancer HO-8910 cells, and the tumor inhibition rates reached 17.03%, 43.3%, and 64.28%, respectively, as shown in Table 6. In the table: ^** P<0.01, vs camellia oil group, t test

Table 6. Therapeutic effect of difetriazole on human ovarian cancer HO-8910 nude mice xenografts

Group Dose Dose Administration Number of Animals Tumor Volume Tumor Weight Tumor Inhibition Rate

(mg/kg) Route (only) (g) (%)

Camellia oil control 10ml/kg im 15 0.507±0.181 0.364±0.124 -

Defetriazole 50.0 im 10 0.182±0.092 ^** 0.130±0.054 ^** 64.28

25.0 im 10 0.352±0.132 ^** 0.207±0.075 ^** 43.13

                                                                                         

Cyclophosphamide 50.0 im 10 0.251±0.207 ^** 0.152±0.042 ^** 58.24

6. Animal pharmacokinetic test of defetriazole

In this experiment, SD rats were selected for pharmacokinetic research, and the dynamic changes of blood concentration of difetriazole in vivo were detected by column switching high performance liquid chromatography, and the curve was fitted on a computer with the 3P87 program of the Chinese Pharmacological Society, and the drug was calculated. Kinetic parameters. The results show that: rat iv ((intravenous injection) administration is a two-compartment kinetics model, and im administration is a one-compartment kinetics model. The pharmacokinetic parameters are respectively:

Rats in iv defetriazole 5, 2.5, 0.5mg/kg three dose groups, average value of 5 rats in each group, t _1/2 α is 0.35, 0.42, 0.40h, t _1/2 β is 2.0, 2.53 , 2.85h, Vd were 1.89, 2.34, 1.95L·kg ^-1 , AUC were 4.12, 1.78, 0.43μg·h·ml ^-1 , Cl(s) were 1.22, 1.42, 1.17L·kg ^-1 ·h ^{- 1} .

Rats im defetriazole 20, 10, 5mg/kg three dose groups, the average value of 5 rats in each group, t _1/2ka is 0.28, 0.35, 0.27h, t _1/2ke is 2.72, 2.33, 2.54h , T(peak) is 0.99, 1.08, 0.93h, Cmax is 1.86, 0.99, 0.55μg·ml ^-1 , AUC is 9.48, 4.65, 2.12μg·h·ml ^-1 , Cl(T) is 2.20, 2.25, 2.78L·kg ^-1 ·h ^-1 , Vd is 8.44, 7.66, 7.68L·kg ^-1 .

Drug distribution test: SD rats, 5 at each time point, were dissected at 1, 8, 24h, 2, 4 and 6d after im defetriazole 5.0 and 50mg/kg respectively, and column switching high performance liquid chromatography was used The distribution of difitriazole in major organs was detected. The results showed that after 1 hour of administration, the order of drug distribution was as follows: uterine ovary > lung > muscle > fat > kidney > small intestine > bone > stomach > liver > heart > spleen > brain; after 48 hours, the order was uterine ovary > lung

>Heart>Kidney>Spleen>Brain>Bone>Muscle>Liver>Fat; after 6 days, the drug concentration was still the highest in uterus and ovary. Drug excretion test: 6 SD rats, iv defetriazole 5mg/kg, measured the excretion of the original drug in urine and feces 1 to 7 days after administration, and also detected by column switching HPLC method, the results showed that: The cumulative excretion in the last 7 days was 2561.7±1968.7ng in urine, accounting for 0.225±0.173% of the administered dose; 1773.2±1335.5ng in feces, accounting for 0.156±0.117% of the administered dose. The amount of unchanged drug excreted in the bile was 0.66±0.13% of the administered dose in 2 hours, and 1.60±0.29% in 12 hours.

Drug-plasma protein binding test: 8 different concentrations of dialysate were used for determination. When the original drug concentration [D]=2×10 ^-6 mol/L, the binding rate of defetriazole to bovine serum albumin was 51.14%.

Bioavailability of difetriazole: $\frac{\mathrm{AUC}\left(\mathrm{im}\right)}{\mathrm{AUC}\left(\mathrm{iv}\right)}\&times;100\%=51.5\%$

7. Acute toxicity test of difetriazole

1). Mouse test:

NIH mice were injected subcutaneously (sc) and intraperitoneally (ip) with triazole camellia oil solution respectively, and respectively established 5 dosage groups, 10 in every group (half and half male and female), and observed the number of deaths within 14 days, according to Bliss's Calculation of LD ₅₀ and reliability. They are: 4385.92 (3979.16-4845.59) mg/kg (sc); 1397.40 (127.21-1514.74) mg/kg (ip).

2) Rat test:

SD rats are the same as the mouse test method, the results of LD ₅₀ and reliability are: 4805.76 (4377.86 ~ 5351.67) mg/kg (sc); 1213.25 (971.67 ~ 1619.01) mg/kg (ip).

8. Long-term toxicity test of defetriazole

1).Rat test:

SD rats were intramuscularly injected (im) three dosage groups of defetriazole 3, 10, and 50mg/kg/d, and solvent camellia oil control group 2ml/kg/d (im), every group of 20 (male and female respectively) half), were administered for 20 days, the results showed that: the high dose group 50mg/kg/d can cause toxic reactions, manifested as bone marrow hematopoietic suppression, thymus atrophy, recovery after 14 days of drug withdrawal, indicating that the toxic reactions are reversible. 10mg/kg/d rats also have mild bone marrow toxicity, and the safe dose of defetriazole without toxic side effects is 3mg/kg/d (im) and below.

2). Rhesus monkey test

Rhesus monkeys (Macaca mulatta) were injected intramuscularly (im) with three dose groups of defetriazole 2, 10, and 50mg/kg/d, and solvent camellia oil control group 2ml/kg/d (im), with 4 animals in each group ( half male and half male), all administered for 20 days, the result was the same as that of rats. 50mg/kg/d can cause toxic reactions, manifested as bone marrow hematopoietic suppression, thymus atrophy,

Recovery after 14 days of drug withdrawal indicated that the toxicity was reversible. 10mg/kg/d also has mild bone marrow toxicity, and the safe dose of non-toxic side effects of rhesus monkey is 2mg/kg/d (im) and below.

3). Beagle dog test

Beagle dogs were injected intramuscularly (im) with three dosage groups of defetriazole 0.05, 0.2, and 0.8mg/kg/d, and solvent camellia oil control group 2ml/kg/d (im), each group of 4 (male and female respectively) half), were administered for three months, and the test results showed that: before and after the administration, the animals of the three dosage groups had respiration, blood pressure, electrocardiogram, and every detection index of blood biochemistry was in the normal range, and there was no physiological significance change. Hematological tests showed that the erythrocytes of animals in the 0.8mg/kg/d and 0.2mg/kg/d dose groups were significantly reduced, but still at a low level of normal, and the rest of the hematological indicators fluctuated within the normal range; gastrointestinal reactions were more common, The 0.8mg/kg/d group had a severe reaction, mainly manifested as anorexia, occasional vomiting, mucus or a small amount of blood in the stool, which disappeared within a short period of time without stopping the drug; The production was inhibited; histopathological examination showed that the bone marrow hematopoietic cells of the animals in the 0.8mg/kg/d group were damaged, the thymus was atrophied, and the immune function was reduced. The above toxic reactions all disappeared after one month of drug withdrawal and returned to normal. Therefore, it can be considered that 0.8 mg/kg/d of difetriazole is the toxic dose for Beagle dogs, and the target sites of toxicity are bone marrow and thymus. 0.2mg/kg/d also has mild bone marrow toxicity, and 0.05mg/kg/d has no obvious gastrointestinal reactions and toxic reactions, which is the basic safe dose for Beagle dogs.

9. Tolerance test of normal people to difetriazole

Phase I clinical trials have shown that the maximum tolerated dose of a single intramuscular injection of defetriazole is 2 mg.

Tests have shown that difetriazole has significant anti-malignant effects, and has a wide anti-tumor spectrum. Through various animal safety tests and normal human tolerance phase I clinical trials, it has been shown that animal species and humans are resistant to difetriazole. Sensitivity varies greatly, and humans and Beagle dogs are most sensitive to defetriazole. The drug has no significant effect on vital signs and is relatively safe. Therefore, it can be considered that difetriazole has clinical prospects in the treatment of malignant tumor diseases.

Detailed ways

Embodiments of the present invention will be described below, but the content of the present invention is not limited thereto at all.

Example 1

Defetriazole Injection

Prescription: Difetriazole 1g, add vegetable oil for injection (tea oil, soybean oil, etc.) to 1000ml.

Preparation: Take vegetable oil for injection, dry heat sterilization at 150°C for 1 hour, let it cool to 50°C, take out part of the oil and add difetriazole, stir to dissolve, and finally add solvent to the full amount. Filter with a dry No. 3 vertical melting funnel, fill in 1ml dry sterilized ampoules, sterilize with flowing steam at 100°C for 30 minutes, and each ampule 1ml contains 1 mg of difetriazole.

Dosage: 0.5-1mg intramuscular injection each time.

Example 2

Triazole nano latex pellets

Dissolve defitriazole in non-aqueous liquid or semi-dry solid compound excipients, prepare soft capsules according to conventional methods, and each pill contains 0.5 mg of defetriazole.

Dosage: 0.5-1mg per oral administration

Once this composite excipient is mixed with water or gastrointestinal fluids, it will rapidly and spontaneously form a nano-sized oil-in-water emulsion, and the drug is dissolved in these extremely small oil droplets and rapidly dispersed to the absorption site in the gastrointestinal tract . During the entire absorption process, the drug remains in a molecular state, and these tiny drug-containing oil droplets can increase its adsorption on the intestinal epithelial cells, prolong the absorption time, increase the transport of the drug through the lymphatic system and pass through the Payer's area of the intestinal tract. Phagocytosis into the body's circulation avoids first-pass elimination. Improved bioavailability and increased drug targeting.

Example 3

Defetriazole Microsphere Injection

Preparation of biocompatible and biodegradable polymers into difetriazole injection microspheres. Each injection contains 2 mg of nontriazole.

Dosage: Intramuscular injection of 2 mg each time, once every 15 days or 30 days.

After intramuscular injection of difetriazole microsphere injection, the polymer will slowly release and absorb the drug at the administration site, prolonging the action time. This polymer is hydrolyzed into harmless degradation products (water and carbon dioxide) after drug release. Intramuscular injection of long-acting non-triazole microsphere injection once can maintain the drug effect for one month. It is used for the prevention of metastasis and recurrence of cancer cells after operation of malignant tumors.

Claims (2)

1. the DPTZ medicament is in preparation treatment ovarian cancer, cervical cancer, lymphoma, leukemia, pulmonary carcinoma, hepatocarcinoma, colorectal cancer, carcinoma of prostate, nasopharyngeal carcinoma, oral cancer, the application in the medicine of pernicious rhabdomyoma disease.

2. purposes according to claim 1 is characterized in that the DPTZ medicament comprises: the DPTZ injection that with the vegetable oil is solvent; The DPTZ microsphere injection liquid; DPTZ nano-emulsion soft gelatin capsule.