Use of 2,3-cyclic GMP-amp sodium salt in preparation of Anti-tumor drug

Abstract

The present invention aims to provide a use of 2,3-Cyclic GMP-AMP sodium salt in the preparation of a drug for treating tumors.

Description

Use of sodium 2,3-cyclic guanosine monophosphate in the preparation of anti-tumor drugs Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to the use of sodium 2,3-cyclic guanosine monophosphate (CGP) in the preparation of anti-tumor drugs. Background Art

[0002] Cancer, also known as malignant neoplasm, is a disease caused by the malfunction of the mechanisms controlling cell growth and proliferation. In addition to uncontrolled growth, cancer cells can also invade surrounding normal tissues and even metastasize to other parts of the body through the circulatory or lymphatic systems.

[0003] Malignant tumors, due to their immature differentiation and rapid growth, can infiltrate and disrupt organ structure and function, and can metastasize, thus severely impacting the body. In addition to causing local compression and obstruction symptoms similar to those of benign tumors, malignant tumors can also present with fever, persistent pain, and, in late stages, severe weight loss, fatigue, anemia, and systemic exhaustion. Ectopic endocrine syndrome: Some non-endocrine gland tumors can produce and secrete hormones or hormone-like substances, causing clinical symptoms of endocrine disorders. These tumors are called ectopic endocrine tumors, and the clinical symptoms they cause are called ectopic endocrine syndrome. These tumors are mostly malignant, most often carcinomas, such as gastric cancer, liver cancer, and colon cancer, but can also occur in sarcomas such as fibrosarcoma and leiomyosarcoma. Furthermore, tumors of the APUD system (diffuse neuroendocrine system), such as carcinoids and pheochromocytomas, can also produce biogenic amines or polypeptide hormones.

[0004] Research on drugs to treat cancer has always been one of the key research directions in the biomedical field.

[0005] Summary of the Invention

[0006] The present invention aims to provide the use of 2,3-cyclic guanosine monophosphate sodium nucleotide in the preparation of anti-tumor drugs.

[0007] The present invention aims to provide the use of 2,3-cyclic guanosine monophosphate sodium nucleotide in the preparation of anti-tumor drugs.

[0008] The present invention aims to provide the use of 2,3-cyclic guanosine monophosphate sodium nucleotide in the preparation of anti-tumor drugs.

[0009] The present invention aims to provide the use of 2,3-cyclic guanosine monophosphate sodium nucleotide in the preparation of anti-tumor drugs.

[0010] The present invention aims to provide the use of 2,3-cyclic guanosine monophosphate sodium nucleotide in the preparation of anti-tumor drugs.

[0011] The present invention aims to provide the use of 2,3-cyclic guanosine monophosphate sodium nucleotide in the preparation of anti-tumor drugs.

[0012] The present invention aims to provide the use of 2,3-cyclic guanosine monophosphate sodium nucleotide in the preparation of anti-tumor drugs.

[0013] 2,3,-Cyclic GMP-AMP sodium salt;

[0014] The pharmacological experiments of the present invention have shown that 2,3-cyclic GMP-sodium nucleotide has a good therapeutic effect on mouse prostate cancer (RM-1 cells), bladder cancer (MB49 cells), breast cancer (4T-1 cells), colon cancer (CT-26 and MC38 cells), and liver cancer (H22 cells). BRIEF DESCRIPTION OF THE DRAWINGS

[0015] The accompanying drawings of the present invention are as follows:

[0016] Figure 1 shows the effect of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide on the tumor growth inhibition of MC38 tumor-bearing mice; in the figure: A is the growth inhibition curve of MC38 tumors after intratumoral administration of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide, and B is the area under the growth inhibition curve of A; C is the growth inhibition curve of MC38 tumors after intraperitoneal administration of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide, and D is the area under the growth inhibition curve of C;

[0017] Figure 2 shows the effect of the test substance 2,3-cyclic GMP-sodium nucleotide on the growth inhibition of MC38 tumor-bearing mice; in the figure: the curve of the inhibition of MC38 tumor growth by intratumoral and intraperitoneal administration of the test substance 2,3-cyclic GMP-sodium nucleotide (A) and the area under the growth inhibition curve-time (AUC) (B);

[0018] Figure 3 shows the effect of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide on the growth inhibition of tumors in H22 tumor-bearing mice; in the figure: A is the growth inhibition curve of H22 tumors after intratumoral administration of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide, and B is the area under the growth inhibition curve of A; C is the growth inhibition curve of H22 tumors after intraperitoneal administration of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide, and D is the area under the growth inhibition curve of C;

[0019] Figure 4 shows the effect of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide on the growth inhibition of tumors in CT-26 tumor-bearing mice; in the figure: A is the growth inhibition curve of CT-26 tumors after intratumoral administration of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide, and B is the area under the growth inhibition curve of A; C is the growth inhibition curve of CT-26 tumors after intraperitoneal administration of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide, and D is the area under the growth inhibition curve of C;

[0020] FIG5 shows the inhibitory effect of different administration routes of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide on tumor growth; ...

[0021] Figure 6 shows the effect of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide on the tumor growth inhibition of MB49 tumor-bearing mice; in the figure: A is the growth inhibition curve of MB49 tumors after intratumoral administration of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide, and B is the area under the growth inhibition curve of A; C is the growth inhibition curve of MB49 tumors after intraperitoneal administration of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide, and D is the area under the growth inhibition curve of C;

[0022] FIG7 shows the effects of different administration routes of the test drug 2,3-cyclic GMP-sodium on tumor growth in MB49 tumor-bearing mice; ...

[0023] Figure 8 shows the effect of the test article 2,3-cyclic guanosine monophosphate-sodium nucleotide on the growth inhibition of tumors in 4T-1 tumor-bearing mice; in the figure: A is the growth inhibition curve of 4T-1 tumors after intratumoral administration of the test article 2,3-cyclic guanosine monophosphate-sodium nucleotide, and B is the area under the growth inhibition curve of A; C is the growth inhibition curve of 4T-1 tumors after intraperitoneal administration of the test article 2,3-cyclic guanosine monophosphate-sodium nucleotide, and D is the area under the growth inhibition curve of C; DETAILED DESCRIPTION

[0024] Example 1 Pharmacological Experiments on the Treatment of Various Cancers with 2,3-Cyclic GMP-Sodium

[0025] 1. Establishment of tumor model mice

[0026] The mixture containing tumor cells was injected into the back of mice, which was recorded as P0 generation. When the volume of P0 generation tumor reached 800mm 3 ~1500mm 3 The mice were killed and the tumor was cut into 3*3*3mm pieces. 3, transplanted to the back of other mice, when the tumor grew to 60-200mm 3 At 14:00, the patients were divided into groups according to the size of the tumors. The average number of tumors in each group was between 80 and 120, and drug administration was started.

[0027] 2. Pharmacodynamics of 2,3-cyclic GMP-sodium nucleotide in different tumor models

[0028] The inhibitory effect of 2,3-cyclic guanosine monophosphate sodium on tumor growth from different tissues at different doses and administration routes was investigated. The number of animals per batch and grouping details are detailed in the main text. Animals weighed 20-30 g. Administration was intratumoral and intraperitoneal. When the average negative control tumor volume reached 1500 mm 3 End the experiment.

[0029] Animal body weight, tumor volume, ascites, and mortality were monitored throughout the experiment. Tumor weight was measured in tumor-bearing mice at the endpoint. Relative tumor volume (TV), relative tumor growth rate (T / C), and tumor inhibition percentage (IR) were calculated and statistically analyzed using SPSS 26.

[0030] Experimental results:

[0031] Tumor inhibition effect of 1,2,3-cyclic guanosine monophosphate sodium on MC38 tumor-bearing mice

[0032] There were two batches of experiments, the first batch was for dose exploration and the second batch was for effectiveness verification.

[0033] Dose-finding experiments:

[0034] Forty-eight MC38 tumor-bearing mice were administered intratumorally and intraperitoneally and divided into 8 groups. The intratumor administration was divided into vehicle control group (Group A), 0.025 mg / kg (Group B), 0.050 mg / kg (Group C), 0.1 mg / kg (Group D), 0.2 mg / kg (Group E), and 0.4 mg / kg (Group F), and the drugs were administered once every 4 days; the intraperitoneal administration group was divided into two dose groups, 20 mg / kg (Group G) and 20 mg / kg (Group H), and the drugs were administered once a day. The experiment lasted for 27 days, and some animals died during the experiment.

[0035] Tumor volume is the observation index:

[0036] Intratumoral administration experiment: As shown in Figure 1A and B, the tumor growth curve shows that after excluding the data of dead animals, the tumor volume of the 0.2 mg / kg and 0.4 mg / kg intratumoral administration groups showed a decreasing trend on the 9th day of administration. At the end of the experiment on the 27th day, the tumor in the model control group was 1451.6±529.3 mm 3The tumor volumes of the test substance 2,3-cyclic GMP-sodium nucleotide administered intratumorally at 0.025 mg / kg (group B), 0.050 mg / kg (group C), 0.1 mg / kg (group D), 0.2 mg / kg (group E), and 0.4 mg / kg (group F) were 1587.6±463.5 mm 3 、1734.3±468.7mm 3 、1340.4±411.8mm 3 、1134.8±526.4mm 3 、484.8±150.6mm 3 The tumor volume inhibition rates were -9.4%, -19.5%, 7.7%, 21.8%, and 66.6%, respectively. All groups achieved the same inhibition rate, except for the 0.4 mg / kg group, which achieved 45% after excluding abnormal data. However, there was a trend toward tumor growth inhibition, suggesting the need for increased dosage. The area under the tumor volume-time curve indicates that sodium 2,3-cyclic GMP-ribonucleotide has a tendency to inhibit tumor growth.

[0037] Intraperitoneal administration experiment: As shown in Figure 1C and D, the tumor growth curve shows that starting from the 15th day after the first administration, the tumor growth of the 20mg / kg intraperitoneal administration group was inhibited compared with the model control group. By the 27th day, the tumor size of the model control group was 1451.6±529.3mm 3 The average tumor volume in the 20 mg / kg and 40 mg / kg dose groups of the test substance 2,3-cyclic GMP-sodium was 483.6±234.5 mm 3 and 783.8±276.0mm 3 The inhibition rates of tumors were 66.7% and 46.0%, respectively, with significant effects (p<0.05).

[0038] Tumor weight is the observation index:

[0039] At the end of the experiment, the tumor weight was weighed and used as an observation indicator.

[0040] Intratumor administration experiment: the drug was given once every 4 days. On the 27th day, the tumor weight of the model control group was 1345.0±581.3 mg, and the tumor weights of the 0.025 mg / kg (Group B), 0.050 mg / kg (Group C), 0.1 mg / kg (Group D), 0.2 mg / kg (Group E), and 0.4 mg / kg (Group F) groups were 1190.0±339.5 mg, 808.3±256.8 mg, 688.3±277.1 mg, 220.7±121.1 mg, and 157.5±71.3 mg, respectively. The tumor inhibition rates were 11.5%, 39.9%, 48.8%, 83.6%, and 88.3%, respectively.

[0041] Intraperitoneal administration: Once daily, on day 27, the model control group had a tumor weight of 1345.0±581.3 mg. A high proportion of mice (5 / 6) died in the 40 mg / kg dose of the test article. The tumor weight in the 20 mg / kg dose group of the test article, 2,3-cyclic GMP-sodium, was 540.0±377.6 mg, with a tumor inhibition rate of 59.9%. The area under the tumor volume-time curve indicates that 2,3-cyclic GMP-sodium can inhibit tumor growth.

[0042] In MC38 tumor-bearing mice, taking tumor volume as an indicator, the effective dose of 2,3-cyclic GMP-sodium nucleotide injected intratumorally is 0.4 mg / kg, 20 mg / kg is effective when administered intraperitoneally, and more animals die at 40 mg / kg, indicating certain toxic side effects.

[0043] 2.2,3-Cyclic GMP-sodium nucleotide inhibition test on MC38 tumor-bearing mice

[0044] Eighteen tumor-bearing mice were divided into three groups: group A, a model control group; group B, a 20 mg / kg intraperitoneal dose, once daily; and group C, a 0.4 mg / kg intratumoral dose, every 4 days. The experiment lasted 17 days, with one mouse death in the low-dose group.

[0045] At the end of the experiment, the negative control tumor volume was 1197.3±385.3mm 3 The tumor volume of groups B and C was 368.7±170.3mm 3 and 877.9±167.1mm 3 Compared with the model control group, the tumor inhibition rate of the test substance 2,3-cyclic GMP-sodium nucleotide was 69.2% when administered intraperitoneally at a dose of 20 mg / kg per day, which was significantly different (P < 0.05). The tumor inhibition rate of the test substance 2,3-cyclic GMP-sodium nucleotide, administered intratumorally once every 4 days at a dose of 0.4 mg / kg, was 26.7%, which did not meet the standard of the preclinical effective dose for tumors, as shown in Figure 2A and B.

[0046] Taking tumor weight as an indicator, the tumor weight of the model control group was 1030.0±272.7 mg, the tumor weight of the intratumoral administration group was 713.3±193.7 mg, and the tumor weight of the intraperitoneal administration group was 443.3±149.0 mg. The tumor inhibition rates relative to the control group were 30.7% and 57.0%, respectively. The body weight of the three groups increased, with percentage changes of 6.6%, 5.3% and 7.6%, respectively. No obvious changes in body weight were observed. The results are shown in Table 1 below.

[0047] Table 1 Effects of the test substance 2,3-cyclic GMP-sodium on tumor weight in MC38 tumor-bearing mice

[0048] Note: a. Mean ± standard error; e. TGI = (mean tumor weight of negative control group - mean tumor weight of treatment group) / mean tumor weight of negative control group * 100%; fT / C% = TTW / CTW × 100% (TTW: mean tumor weight of treatment group at the end of the experiment; CTW: mean tumor weight of negative control group at the end of the experiment). Tumor volume on day 15 of gc was compared between the treatment group and the control group using one-way analysis of variance. *P < 0.05 vs. model control group.

[0049] 3. Tumor inhibition of the test drug 2,3-cyclic GMP-sodium on H22 tumor-bearing mice

[0050] Forty-eight tumor-bearing mice received intratumoral and intraperitoneal administration of the drug. Group E received 400 μg / kg intratumorally every four days, while Group F received 800 μg / kg intratumorally. Intraperitoneal administration consisted of vehicle (Group A), 10 mg / kg (Group B), 20 mg / kg (Group C), and 40 mg / kg (Group D). The experiment lasted for 16 days, with no mouse deaths.

[0051] Tumor volume was used as the observation index:

[0052] Intratumoral administration experiment: From the tumor growth curve, starting from the 12th day after the first administration, the high-dose group of intratumoral administration showed a significant inhibition of tumor growth compared with the model control group. By the 16th day, the tumor in the model control group reached 3505.0±1272.3mm 3 The tumor volumes of group F and group B after intratumoral administration of 400ug / kg and 800ug / kg of the test substance 2,3-cyclic GMP-sodium nucleotide were 1936.1±2332.6mm 3 and 473.7±497.9mm 3 Compared with the control group, the high-dose group administered intratumorally significantly inhibited tumor growth (p < 0.001) (Figure 3A). The area under the tumor growth volume-time curve (AUC) of the high-dose group of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide was 2530.0 ± 672.2 mm 3 .d was significantly lower than that of the model control group (AUC 12979.0±4879.0mm 3 .d(p<0.01), see Figures 3A and B.

[0053] Intraperitoneal administration experiment: From the tumor growth curve, starting from the 8th day after the first administration, the tumor growth of each administration group was significantly inhibited compared with the model control group. By the 16th day, the tumor in the model control group reached 3505.0±1272.3mm3 The tumor volumes of the test substance 2,3-cyclic GMP-sodium were 81.5±24.5, 40.6±14.5 and 32.5±9.3 mm2 at 10 mg / kg Qd(ip) (group B), 20 mg / kg Qd(ip) (group C) and 40 mg / kg Qd(ip) (group D), respectively. 3 The tumor volume-time curve area in the model control group was 12979.0±4879.0mm. 3 .d, the AUC of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide in the 10, 20 and 40 mg / kg dose groups were 1609.0±235.1, 1408.0±352.3 and 1163.0±193.2 mm 3 .d, significantly lower than that in the model control group (see Figure 3C and D).

[0054] Tumor weight was used as the observation index:

[0055] At the end of the experiment, the tumor weight was weighed and used as an observation indicator.

[0056] Intratumoral administration experiment: The tumor weight of the model control group was 3558.6±959.8 mg, and the tumor weight of the test sample 2,3-cyclic guanosine monophosphate-sodium 400ug / kg and 800ug / kg groups was 2138.3±690.4 and 371.7±89.7 mg, respectively. The tumor inhibition rates were 39.9% and 89.6%, respectively, and the inhibitory effect on the tumor was significant (p<0.05 or p<0.001).

[0057] Intraperitoneal administration experiment: The tumor weight of the model control group was 3558.6±959.8 mg, and the tumor weights of the test article 2,3-cyclic guanosine monophosphate-sodium 10, 20, and 40 mg / kg groups were 100.0±19.4, 205.0±47.1, and 0.0±0.0 mg. The tumor inhibition rates were 97.2%, 94.2%, and 100.0%, respectively. The inhibitory effect on the tumor was significant (p<0.05 or p<0.001). The results are shown in Table 2 below.

[0058] Taking weight as observation indicator:

[0059] At the end of the experiment, the body weight of the animals after administration decreased compared to the model control group. Using the same administration route, the control group experienced a 22.1% increase in weight compared to the initial body weight. The body weight gain of the test article, 2,3-cyclic GMP-sodium, was negatively correlated with the dose administered. The body weight change rates for intratumoral administration were 14.3% and 10.3%, respectively; and for intraperitoneal administration, the body weight change rates were 5.7%, 7.8%, and -0.8%, respectively.

[0060] In addition, the animals in each group of intraperitoneal administration developed ascites to varying degrees, while the animals in each group of intratumoral administration did not develop ascites.

[0061] The above experiment was repeated 3 times, and the change trend was consistent.

[0062] Table 2 Effects of the test substance 2,3-cyclic GMP-sodium on tumor weight in H22 tumor-bearing mice

[0063] Note: a. Mean ± standard error; e. TGI = (mean tumor weight of negative control group - mean tumor weight of treatment group) / mean tumor weight of negative control group * 100%; fT / C% = TTW / CTW × 100% (TTW: mean tumor weight of treatment group at the end of the experiment; CTW: mean tumor weight of negative control group at the end of the experiment). The tumor volume on day 15 of gc was compared between the treatment group and the control group by one-way analysis of variance, *P < 0.05, **P < 0.01, ***P < 0.01 vs. model control group.

[0064] 4. Tumor inhibition of the test drug 2,3-cyclic GMP-sodium on CT-26 tumor-bearing mice

[0065] Thirty-six CT26 tumor-bearing mice were given intratumoral and intraperitoneal administration. The intratumoral administration groups were divided into vehicle control group (intramedullary, group A), 0.2 mg / kg (intramedullary, group B), 1 mg / kg (intramedullary, group C), and 5 mg / kg (intramedullary, group D), and the drug was administered once every two days. The intraperitoneal administration groups were: 5 mg / kg (ip, group E), and 10 mg / kg (ip, group F). The experiment lasted for 14 days, and no animals died during the experiment.

[0066] Tumor volume was used as the observation index:

[0067] Intratumoral administration experiment: From the tumor growth curve, starting from the 10th day after the first administration, the high-dose group of intratumoral administration showed a significant inhibition of tumor growth compared with the model control group. By the 14th day, the tumor size of the model control group was 1554.0±429.1mm 3 The tumor volumes of the test substance 2,3-cyclic GMP-sodium nucleotide in the 0.2 mg / kg, 1 mg / kg, and 5 mg / kg groups were 847.6±166.2, 606.8±192.9, and 204.1±133.5 mm 3 Compared with the control group, each intratumoral administration group significantly inhibited tumor growth in a dose-dependent manner (p<0.001). The area under the tumor volume-time curve also showed that 2,3-cyclic GMP-sodium inhibited tumor growth in a dose-dependent manner, as shown in Figure 4A and B.

[0068] Intraperitoneal administration experiment: From the tumor growth curve, starting from the 12th day after the first administration, the tumor growth of the 20mg / kg intraperitoneal administration group was significantly inhibited compared with the model control group. By the 16th day, the tumor of the model control group reached 1554.0±429.1mm 3 The 5mg / kg and 20mg / kg dose groups of the test substance 2,3-cyclic guanosine monophosphate-sodium nucleotide were 1246.0±178.2 and 884.1±275.6mm 3 The high-dose group had a significant inhibitory effect on tumors (p<0.01), as shown in Figure 4C and D.

[0069] Tumor weight was used as the observation index:

[0070] At the end of the experiment, the tumor weight was weighed and used as an observation indicator.

[0071] Intratumoral administration experiment: The tumor weight of the model control group was 1229.2±453.8 mg, and the tumor weight of the test article 2,3-cyclic guanosine monophosphate-sodium 0.2 mg / kg, 1 mg / kg, and 5 mg / kg groups were 500.6±173.5, 357.5±139.9, and 106.2±93.5 mg. The tumor inhibition rates were 59.3%, 70.9%, and 91.4%, respectively, and the inhibitory effect on tumors was significant (p<0.05 or p<0.001).

[0072] Intraperitoneal administration experiment: The tumor weight of the model control group was 1229.2±453.8 mg, and the tumor weight of the test article 2,3-cyclic guanosine monophosphate-sodium 5 and 20 mg / kg groups was 1036.2±188.2 mg and 648.0±166.0 mg, respectively. The tumor inhibition rates were 15.7% and 47.3%, respectively. The test article 2,3-cyclic guanosine monophosphate-sodium 20 mg / kg group had a significant inhibitory effect on the tumor (p<0.01), see Table 3 below.

[0073] Taking weight as observation indicator:

[0074] At the end of the experiment, the body weight of the animals after administration decreased compared to the model control group. Using the same administration method, the control group's body weight increased by 13.4% compared to the initial body weight. Intratumoral administration of the test article, 2,3-cyclic GMP-sodium nucleotide, showed little change in body weight, with the rates of change being 12.9%, 13.1%, and 10.1%, respectively. The rates of change after intraperitoneal administration were 12.0% and 2.3%, respectively.

[0075] Effect of drug administration route on tumor growth

[0076] Under the same administration route, intratumoral administration was significantly better than intraperitoneal administration in inhibiting tumor growth, as shown in Figure 5A and B.

[0077] In addition, the animals in each group of intraperitoneal administration developed ascites to varying degrees, while the animals in each group of intratumoral administration did not develop ascites.

[0078] The above experiment was repeated 4 times, and the change trend was consistent.

[0079] Table 3 Effects of the test substance 2,3-cyclic GMP-sodium on tumor weight in CT-26 tumor-bearing mice

[0080] Note: a. Mean ± standard error; e. TGI = (mean tumor weight of negative control group - mean tumor weight of treatment group) / mean tumor weight of negative control group * 100%; fT / C% = TTW / CTW × 100% (TTW: mean tumor weight of treatment group at the end of the experiment; CTW: mean tumor weight of negative control group at the end of the experiment). The tumor volume on day 15 of gc was compared between the treatment group and the control group by one-way analysis of variance, *P < 0.05, **P < 0.01, ***P < 0.01 vs. model control group.

[0081] 5. Tumor inhibition of 2,3-cyclic GMP-sodium nucleotide on MB49 tumor-bearing mice

[0082] Thirty MB49 tumor-bearing mice were given intratumoral and intraperitoneal administration. The intratumoral administration groups were divided into vehicle control group (Group A), 0.3 mg / kg (Group B), and 3 mg / kg (Group C), and were administered once every 2 days; the intraperitoneal administration groups were divided into 3 mg / kg (Group D), 10 mg / kg (Group E), and 20 mg / kg (Group F), and were administered once a day. The experiment lasted for 14 days, and no animals died during the experiment.

[0083] Tumor volume was used as the observation index:

[0084] Intratumoral administration experiment: From the tumor growth curve, starting from the 12th day after the first administration, the high-dose group of intratumoral administration showed a significant inhibition of tumor growth compared with the model control group. By the 14th day, the tumor size of the model control group was 5975.6±936.8mm 3 The tumor volume of the test substance 2,3-cyclic GMP-sodium nucleotide group administered intratumorally at 0.3 mg / kg and 3.0 mg / kg was 3409.0±991.0 mm 3 and 2656.4±742.8mm 3 Compared with the control group, the high-dose intratumoral administration group significantly inhibited tumor growth (p<0.05). The area under the tumor volume-time curve showed that 2,3-cyclic GMP-sodium can inhibit tumor growth, as shown in Figures 6A and B.

[0085] Intraperitoneal administration experiment: From the tumor growth curve, starting from the 10th day after the first administration, the 20mg / kg intraperitoneal administration group had a significant inhibitory effect on tumor growth compared with the model control group. By the 16th day, the tumor size of the model control group was 5975.6±936.8mm 3 The test sample 2,3-cyclic guanosine monophosphate-sodium nucleotide in the 3mg / kg, 10mg / kg, and 20mg / kg dose groups were 2656.4±742.8mm 3 、3384.1±921.5mm 3 and 1365.8±174.5mm 3 The high-dose group had a significant inhibitory effect on tumors (p<0.01), as shown in Figures 6C and D.

[0086] Tumor weight was used as the observation index:

[0087] At the end of the experiment, the tumor weight was weighed and used as an observation indicator.

[0088] Intratumoral administration experiment: The tumor weight of the model control group was 4401.9±618.3 mg, and the tumor weight of the test article 2,3-cyclic guanosine monophosphate-sodium 0.3 mg / kg and 3.0 mg / kg groups was 3290±1156.3 mg and 1398.2±563.6 mg, respectively. The tumor inhibition rates were 25.3% and 68.2%, respectively. The high-dose group had a significant inhibitory effect on the tumor (p<0.05).

[0089] Intraperitoneal administration experiment: The tumor weight of the model control group was 4401.9±618.3 mg, and the tumor weights of the test article 2,3-cyclic guanosine monophosphate-sodium nucleotide 3 mg / kg, 10 mg / kg, and 20 mg / kg dose groups were 2356.6±420.0, 2394.5±1080.2, and 826.1±156.1 mg, respectively. The tumor inhibition rates were 46.5%, 45.6%, and 81.2%, respectively. The test article 2,3-cyclic guanosine monophosphate-sodium nucleotide 20 mg / kg group had a significant inhibitory effect on the tumor (p<0.01), as shown in Table 4.

[0090] Taking weight as observation indicator:

[0091] At the end of the experiment, the body weight of the animals after administration was no different from that of the model control group. The body weight of the animals in all groups increased, with the increase percentages in groups A to F being 18.6%, 20.4%, 13.9%, 19.2%, 19.8% and 13.1% respectively.

[0092] Effect of administration route on tumor growth:

[0093] Under the same administration route, intratumoral administration tends to be more effective in inhibiting tumor growth than intraperitoneal administration, as shown in Figures 7A and 7B.

[0094] In addition, the animals in each group of intraperitoneal administration developed ascites to varying degrees, while the animals in each group of intratumoral administration did not develop ascites.

[0095] The above experiment was repeated once, and the change trend was consistent.

[0096] Table 4 Effects of the test substance 2,3-cyclic GMP-sodium on tumor weight in MB49 tumor-bearing mice

[0097] Note: a. Mean ± standard error; e. TGI = (mean tumor weight of negative control group - mean tumor weight of treatment group) / mean tumor weight of negative control group * 100%; fT / C% = TTW / CTW × 100% (TTW: mean tumor weight of treatment group at the end of the experiment; CTW: mean tumor weight of negative control group at the end of the experiment). The tumor volume on day 15 of gc was compared between the treatment group and the control group using one-way analysis of variance. *P < 0.05, **P < 0.01 vs. model control group.

[0098] 6. Tumor inhibition of 2,3-cyclic guanosine monophosphate sodium on 4T-1 tumor-bearing mice

[0099] Thirty MB49 tumor-bearing mice were given intratumoral and intraperitoneal administration. The intratumoral administration was divided into a solvent control group (Group A), 0.1 mg / kg (Group B), 0.3 mg / kg (Group C), and 1 mg / kg (Group D), and the drug was administered once every 2 days; the intraperitoneal administration group set up a 10 mg / kg group (Group E), and the drug was administered once a day. The experiment lasted for a total of 14 days, and no animals died during the experiment.

[0100] Tumor volume is the observation index:

[0101] Intratumoral drug administration experiment: From the tumor growth curve, on the 16th day, the tumor size of the model control group was 1636.5±261.8mm 3 The tumor volume of the test substance 2,3-cyclic GMP-sodium nucleotide in the 0.1 mg / kg (group B), 0.3 mg / kg (group C), and 1 mg / kg (group D) groups was 1481.5±182.6 mm 3 、1052.9±148.4mm 3 、1044.7±251.2mm 3 , none of which achieved a 45% tumor inhibition rate, indicating a trend toward tumor growth inhibition, suggesting the need for increased dosage. The area under the tumor volume-time curve indicates that sodium 2,3-cyclic GMP-ribonucleotide can inhibit tumor growth (see Figures 8A and 8B).

[0102] Intraperitoneal administration experiment: From the tumor growth curve, starting from the 4th day after the first administration, the tumor growth of the 20mg / kg intraperitoneal administration group was inhibited compared with the model control group. By the 16th day, the tumor size of the model control group was 1636.5±261.8mm 3 The tumor volume of the test substance 2,3-cyclic GMP-sodium at a dose of 10 mg / kg was 891.8±118.1 mm 3 , and the inhibitory effect on tumors was significant (p<0.05), as shown in Figures 8C and 8D.

[0103] Tumor weight was used as the observation index:

[0104] At the end of the experiment, the tumor weight was weighed and used as an observation indicator.

[0105] Intratumoral administration experiments: The tumor weight in the model control group was 1490.0±610.3 mg, while the tumor volumes in the 0.1 mg / kg (Group B), 0.3 mg / kg (Group C), and 1 mg / kg (Group D) groups were 1441.7±469.1 mg, 825.0±526.8 mg, and 1121.7±732.1 mg, respectively. The tumor inhibition rates were 3.24%, 44.63%, and 24.72%, respectively, falling short of the 45% tumor inhibition rate. This suggests a trend toward tumor growth inhibition, suggesting the need for increased dosage. The area under the tumor volume-time curve indicates that sodium 2,3-cyclic GMP-ribonucleotide can inhibit tumor growth.

[0106] Intraperitoneal administration: The tumor weight in the model control group was 1490.0±610.3 mg, and the tumor weight in the 20 mg / kg 2,3-cyclic GMP-sodium group was 1138.3±382.5 mg. The tumor inhibition rates were 23.60%, respectively. The tumor inhibition trend of the 20 mg / kg 2,3-cyclic GMP-sodium group is shown in Table 5.

[0107] Taking weight as observation indicator:

[0108] At the end of the experiment, the body weight of the animals after administration was no different from that of the model control group. The body weight of the animals in all groups increased, with the increase percentages in groups A to E being 7.6%, 2.6%, 9.1%, -0.5% and -0.7%.

[0109] In addition, the animals in each group of intraperitoneal administration developed ascites to varying degrees, while the animals in each group of intratumoral administration did not develop ascites.

[0110] Table 5 Effects of the test substance 2,3-cyclic GMP-sodium on tumor weight in 4T-1 tumor-bearing mice

[0111] Note: a. Mean ± SD; e. TGI = (mean tumor weight of negative control group - mean tumor weight of treatment group) / mean tumor weight of negative control group * 100%; fT / C% = TTW / CTW × 100% (TTW: mean tumor weight of treatment group at the end of the experiment; CTW: mean tumor weight of negative control group at the end of the experiment). Tumor volume on day 15 of gc was compared between the treatment group and the control group using one-way ANOVA. *P < 0.05 vs. model control group.

[0112] 7. Tumor inhibition of RM-1 tumor-bearing mice by sodium 2,3-cyclic guanosine monophosphate (CGP)

[0113] The experiment was conducted in three batches. Because the tumor grew too fast, one of the batches performed better.

[0114] A total of 18 tumor-bearing mice were divided into three groups: model control group (Group A), test article BJY-0.3mg / kg (Group B), and 1mg / kg (Group C). All were administered intratumorally. The experiment lasted 12 days, and two mice in the high-dose group died.

[0115] At the end of the experiment, the negative control tumor volume was 7440.0±1262.8mm 3 The tumor volume of groups B and C was 3946.0±843.0mm 3 (P<0.05) and 3527.6±962.4mm 3 The tumor inhibition rates of the model control group were 47.0% and 52.6%.

[0116] The tumor weight in the model control group was 6300.0±1105.2 mg. The tumor weights in the 0.3 mg / kg and 1 mg / kg intratumoral administration groups were 2780.0±603.4 mg and 2520.0±547.6 mg, respectively. These represent tumor inhibition rates of 56% and 60%, respectively, relative to the model control group. Body weight changes decreased with increasing and decreasing doses, with rates of change of 31%, 27%, and 25%, respectively. See Table 6.

[0117] Table 6 Effect of the test substance 2,3,-cyclic GMP-sodium on tumor weight in RM-1 tumor-bearing mice

[0118] Note: a. Mean ± standard error; e. TGI = (mean tumor weight of negative control group - mean tumor weight of treatment group) / mean tumor weight of negative control group * 100%; fT / C% = TTW / CTW × 100% (TTW: mean tumor weight of treatment group at the end of the experiment; CTW: mean tumor weight of negative control group at the end of the experiment). The tumor volume on day 15 of gc was compared between the treatment group and the control group using one-way analysis of variance, *P < 0.05 vs. model control group.

Claims

1. Use of sodium 2,3-cyclic guanosine monophosphate (CGP) in the preparation of drugs for treating tumors.

2. Use of sodium 2,3-cyclic guanosine monophosphate (CGP) in the preparation of drugs for treating prostate cancer.

3. Use of 2,3'-cyclic guanosine monophosphate sodium nucleotide in the preparation of drugs for treating bladder cancer.

4. Use of 2,3'-cyclic guanosine monophosphate-sodium nucleotide in the preparation of drugs for treating breast cancer.

5. Use of 2,3'-cyclic guanosine monophosphate sodium nucleotide in the preparation of drugs for treating colon cancer.

6. Use of sodium 2,3'-cyclic guanosine monophosphate-nucleotide in the preparation of drugs for treating liver cancer.

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