Application of icaritin in preparation of medicine for treating and / or relieving pain

Abstract

The invention relates to an application of icaritin in preparation of a medicine for treating and / or relieving pain, especially tumor-related pain.

Description

Technical Field

[0001] The present invention relates to the use of icariin in the preparation of a medicament for treating and / or alleviating pain, and belongs to the field of medicine. Background Art

[0002] When malignant tumors progress to the advanced stage, due to the large size of the tumors and their invasion of surrounding tissues, pain symptoms will occur. Bone metastatic cancer pain is one of the most common complications of cancer, and its incidence increases year by year with the overall prevalence of cancer, seriously affecting the quality of life and survival period of cancer patients. Bone metastatic cancer pain is a special type of pain that combines the characteristics of inflammatory pain and neuropathic pain but is different from both types, and its pathogenesis is very complex.

[0003] Bone cancer pain (BCP) is pain caused by tumor metastasis to the bone, which often occurs in breast cancer, prostate cancer, lung cancer, etc. It is found that more than 80% of advanced cancer patients clinically experience varying degrees of bone pain, seriously affecting the quality of life of patients. The pathogenesis of BCP is complex and involves physiological and pathological factors. On the one hand, tumor cells activate osteoclasts, leading to bone destruction and causing bone pain. On the other hand, with the growth of the tumor and bone destruction, tumor cells and related stromal cells secrete various cytokines, and the bone marrow constitutes a complex and unique microenvironment, thus activating and sensitizing the nociceptors that innervate bone tissue. These peripheral nociceptive information is transmitted through spinal neurons and uploaded to the central nervous system, and through the interaction between neurons and glial cells, synaptic remodeling, and activation of the descending facilitation system, etc., it leads to pain hypersensitivity and / or hyperalgesia, ultimately resulting in BCP.

[0004] Clinical treatment of BCP includes various treatment methods such as three-step analgesic drugs, bisphosphonates, interventional therapy, bone protectants, palliative radiotherapy, surgery, and anti-tumor therapy, which play a certain role in controlling BCP. However, these treatment measures all have some problems. For example, non-steroidal anti-inflammatory drugs (NSAIDs) can inhibit prostaglandin synthesis and damage the digestive tract mucosal barrier, so digestive tract adverse reactions often occur during the application of NSAIDs, including dyspepsia, digestive tract ulcers, digestive tract bleeding, etc. And there is a problem of the maximum effective dose (ceiling effect) for NSAIDs and weak opioid drugs. Although strong opioid analgesic drugs do not have a ceiling effect and have obvious analgesic effects, they are prone to produce toxic and side effects such as drug resistance, addiction, constipation, drowsiness, and respiratory depression after long-term large-dose administration; bisphosphonate drugs can cause osteonecrosis of the jaw and influenza-like symptoms, while radiotherapy and chemotherapy are prone to cause bone marrow suppression, and these adverse reactions often affect or even interrupt the treatment of BCP.

[0005] Primary bone tumors are divided into benign tumors and malignant tumors, and the malignant tumors are called bone cancer. It includes various types such as malignant chondrogenic tumors, malignant osteogenic tumors, malignant fibrogenic tumors, malignant bone tumors of the hematopoietic system, malignant angiosarcoma, malignant osteoleiomyosarcoma, malignant bone lipogenic tumors, etc. Chemotherapy drugs for the clinical treatment of primary bone cancer generally include high-dose methotrexate, ifosfamide, cisplatin, doxorubicin, cyclophosphamide, vincristine, actinomycin, etoposide, doxorubicin and ifosfamide, etc. Chemotherapy can control tumors, but it cannot relieve pain. The reason is that after cells rupture or are damaged during chemotherapy, the original substances inside the cells are released to the outside of the cells, resulting in an increase in the content of local inflammatory mediators and potassium ions, which causes the pain to intensify; moreover, chemotherapy drugs can cause serious adverse reactions.

[0006] Methotrexate is an antimetabolic drug that can inhibit DNA synthesis and cell replication. Methotrexate can cause mucosal damage, liver function damage, bone marrow suppression and subacute neurotoxicity.

[0007] Cyclophosphamide is a kind of alkylating agent among the anti-cancer chemotherapy drugs, and the main administration route is intravenous injection. After injection, the blood vessels at the injection site of the patient first turn red, painful and spread proximally. In severe cases, there may be brachial plexus neuralgia. The patient shows profuse sweating and groans continuously. Two weeks later, the blood vessels become significantly blackened and hardened, showing a cord-like shape.

[0008] Ifosfamide can produce central neurotoxicity, and the clinical manifestations are hallucinations, delirium, apathy and anxiety. In severe cases, there are manifestations such as epileptic seizures and even coma; and ifosfamide causes cardiac toxicity, and the clinical manifestations are heart failure and malignant arrhythmia. Since the side effects of traditional drugs for treating pain related to primary and secondary bone cancer are obvious, a drug with less side effects is needed to solve the pain related to bone cancer and even other tumors.

[0009] Icaritin, also known as acoradin and icariin aglycone, is a new effective monomer obtained by enzymatic transformation of icariin, the main active ingredient isolated from the traditional Chinese medicine Epimedium brevicornu Maxim. Its structural formula is shown in the following formula (I):

[0010]

[0011] The use of icariin in the preparation of drugs for abnormal cell proliferation, especially cancer, was mentioned in the Chinese patent application with the application number 200780039276.9. The application of icariin in the preparation of selective estrogen receptor modulators was proposed in the Chinese patent with the application number 03129242.9. The use of icariin in the treatment of tumors has been disclosed in the existing documents, but there is no report on the pain related to tumors. Summary of the Invention

[0012] One object of the present invention is to provide the use of icariin in the preparation of a medicament for treating and / or alleviating pain.

[0013] On the one hand, the present invention provides the use of icariin in the preparation of a medicament for treating and / or alleviating pain.

[0014] Preferably, the pain includes tumor-related pain.

[0015] Preferably, the tumor includes solid tumors.

[0016] Preferably, the tumor-related pain includes pain associated with bone cancer.

[0017] Preferably, the pain associated with bone cancer includes pain related to primary bone cancer and secondary bone cancer.

[0018] Preferably, the primary bone cancer includes chondrosarcoma, osteogenic tumor and osteofibroma.

[0019] Preferably, the secondary bone cancer includes secondary bone cancer associated with one or more of breast cancer, lung cancer, prostate cancer, thyroid cancer, liver cancer, pancreatic cancer and colorectal cancer bone metastasis.

[0020] Preferably, the solid tumors include one or more of breast cancer, lung cancer, prostate cancer, thyroid cancer, liver cancer, pancreatic cancer and colorectal cancer.

[0021] Preferably, the medicament is an oral preparation.

[0022] Preferably, the oral preparation is one or more of capsules, tablets, solutions, syrups, emulsions, suspensions and granules.

[0023] The effect of the present invention is that the icariin of the present invention shows good therapeutic effects in the use of medicaments for treating and / or alleviating pain, especially tumor-related pain. This includes pain related to primary bone cancer and secondary bone cancer. Compared with existing medicaments for treating tumor-related pain, the icariin of the present invention has fewer side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 Indicates mechanical hyperalgesia in mice of the blank group, phosphate buffer group and tumor model group.

[0025] Figure 2 Indicates cold hyperalgesia in mice of the blank group, phosphate buffer group and tumor model group.

[0026] Figure 3 Indicates spontaneous pain in mice of the blank group, phosphate buffer group and tumor model group.

[0027] Figure 4 Indicates the mechanical hyperalgesia of mice after inoculation with LLC cells and administration of different doses of icariin to the mice.

[0028] Figure 5 Indicates the cold hyperalgesia of mice after inoculation with LLC cells and administration of different doses of icariin to the mice.

[0029] Figure 6 Indicates the mechanical hyperalgesia of mice after administration of different doses of icariin to the mice for 7 days after administration.

[0030] Figure 7 Indicates the cold hyperalgesia of mice after administration of different doses of icariin to the mice for 7 days after administration.

[0031] Figure 8 Indicates the mechanical hyperalgesia of mice after administration according to the method of Example 3.

[0032] Figure 9 Indicates the cold hyperalgesia of mice after administration according to the method of Example 3.

[0033] Figure 10 Indicates the number of spontaneous foot lifts of mice within 10 minutes after administration according to the method of Example 3.

[0034] Figure 11 Indicates the foot lift protection time of mice within 10 minutes after administration according to the method of Example 3. Detailed implementation manners

[0035] The following examples are only used for exemplary illustration of the present invention and are not used to limit the present invention. Modifications, changes, variations, etc. made within the protection scope of the present invention are all within the protection scope of the present invention.

[0036] Unless otherwise stated, "mechanical hyperalgesia" herein refers to the pain that occurs in body tissues under the action of external forces. When body tissues are subjected to external force stimulation, they will deform. When the degree of deformation exceeds the threshold of mechanoreceptors, the nociceptors are activated, thereby causing the body to produce a feeling of pain.

[0037] Unless otherwise stated, "cold hyperalgesia" herein refers to the pain that occurs when the body's sensitivity to cold exceeds the threshold of thermoreceptors after body tissues are damaged, and the nociceptors are activated, thereby causing the body to produce a feeling of pain.

[0038] Unless otherwise stated, "spontaneous pain" herein refers to the pain that spontaneously occurs inside the body without any external stimulation.

[0039] Unless otherwise stated, "icariin soft capsules" herein are marketed drugs and can be purchased from hospitals.

[0040] Unless otherwise specified, the "Lewis lung cancer cells" in this article were purchased from Shanghai Zeye Biotechnology Co., Ltd., product number: CB95529855.

[0041] Example 1

[0042] Evaluation of the analgesic effect of icariin at different doses on mice with bone cancer pain

[0043] I. Experimental animals

[0044] 8-week-old male C57BL / 6J littermate mice, weighing 20 - 22 g.

[0045] II. Bone cancer pain modeling cells

[0046] Lewis lung cancer cells (abbreviated as LLC cells), 2×10 5 cells were used for each mouse.

[0047] III. Experimental methods

[0048] 1. Establishment of bone cancer pain model mice:

[0049] (1) Preparation of LLC cell suspension

[0050] Resuscitate LLC cells. When the cells are almost confluent in the culture dish, perform one or two passages to ensure the state and quantity of tumor cells. After washing the culture dish 2 times with Dulbecco's phosphate buffered saline (DPBS), digest the cells with 0.25% trypsin for 5 minutes. After terminating the digestion with complete medium, transfer the cells to a 15 ml centrifuge tube, centrifuge at 1000 rpm for 3 minutes, discard the supernatant, resuspend the cells with phosphate buffered saline (PBS), and place them in an ice box for later use.

[0051] (2) Grouping of experimental mice

[0052] 1.2.1 Establishment of bone cancer pain mouse model:

[0053] Modeling surgery

[0054] Anesthetize the mice with isoflurane gas for 2 minutes or with 1% sodium pentobarbital, inject the anesthetic intraperitoneally at a dose of 0.05 g / kg. Then place the mice on the surgical drape, disinfect the skin of the left thigh and joint of the mice, and use a 10 μl microsyringe to aspirate the corresponding volume of tumor cell suspension from the LLC cell suspension. Inject approximately 2×10 5 LLC cells into the bone marrow cavity of the distal end of the left femur of the mice and stop the needle for 2 minutes. After injection, seal the wound with bone wax and finally suture the wound.

[0055] 1.2.2 Blank mouse model group

[0056] No operations are performed on the mice.

[0057] 1.2.3 Phosphate Buffered Saline (PBS) model group

[0058] Anesthetize the mice with isoflurane gas for 2 minutes or with 1% sodium pentobarbital (0.05 g / kg, intraperitoneal injection). Then place the mice on the surgical drape, disinfect the skin of the left thigh and joint of the mice, use a 10 μl micro syringe to aspirate 10 ml of 0.01 M PBS and inject it into the bone marrow cavity of the distal left femur of the mice, and stop the needle for 2 minutes. After injection, seal the wound with bone wax and finally suture the wound.

[0059] 2. Behavioral determination:

[0060] (1) Mechanical pain determination: In a quiet environment, place the mice in a plastic compartment with a wire mesh bottom. After 30 minutes of adaptation, start the measurement. Apply von Frey filaments (0.02, 0.04, 0.07, 0.16, 0.4, 0.6, 1.0, and 2.0 g) vertically to the center of the left hind paw of the mice and hold for 5 seconds. A sudden withdrawal, licking, or shaking of the foot in response to von Frey filament stimulation is considered a positive reaction. Each filament of each gram is measured 5 times, with an interval of at least 10 minutes between each measurement. The minimum gram required to produce 3 positive reactions is recorded as the mechanical pain threshold.

[0061] (2) Spontaneous pain determination: In a quiet environment, place the mice in a plastic compartment with a glass platform bottom. After 30 minutes of adaptation, start the measurement. Then, without any stimulation, record the number and duration of spontaneous lifting, licking, shaking, or limping of the left foot of the mice within 10 minutes as the judgment indicators of spontaneous pain behavior. Among them, the number of spontaneous lifting, licking, shaking, or limping of the left foot of the mice within 10 minutes is called the "number of foot lifts within 10 minutes", and the duration is called the "foot lift protection time within 10 minutes".

[0062] (3) Determination of cold pain sensitivity: The cold stimulation behavioral device is used to measure the cold pain latency. In a quiet environment, place the mice in a plastic compartment with a glass platform bottom. After 30 minutes of adaptation, start the measurement. Place the instrument with dry ice under the glass platform and vertically contact the glass under the left hind paw of the mice. The longest time of dry ice contact is set to 20 seconds to prevent unnecessary tissue damage. A sudden withdrawal, licking, or shaking of the foot in response to cold stimulation is considered a positive reaction. Each mouse is measured 3 times, with an interval of at least 10 minutes between each measurement. Record the time required for a positive reaction, and the average of the 3 results is recorded as the cold pain "withdrawal latency threshold".

[0063] The above experimental methods can also refer to the experimental methods in the priority document 202311330329.X.

[0064] The mechanical hyperalgesia, cold hyperalgesia and spontaneous pain curves of the mice in the tumor model group, blank group and phosphate buffer group are shown in Figure 1 , Figure 2 and Figure 3 .

[0065] Example 2

[0066] The bone cancer animal model of Example 1 was divided into the following groups:

[0067] Group 1: The group given corn oil (solvent) by gavage, that is, the control group at 0 mg / kg;

[0068] Group 2: The group given icariin at a dose of 50 mg / kg each time by gavage, that is, experimental group 1;

[0069] Group 3: The group given icariin at a dose of 100 mg / kg each time by gavage, that is, experimental group 2;

[0070] Group 4: The group given icariin at a dose of 200 mg / kg each time by gavage, that is, experimental group 3;

[0071] Group 5: The group given icariin at a dose of 400 mg / kg each time by gavage, that is, experimental group 4.

[0072] There were 11 - 15 experimental animals (bone cancer pain model mice) in each group. Administration method: gavage (the day of modeling was day 0, and the drug was administered on the 7th day after modeling, 2 times a day), and the drug was administered continuously for 7 days until the 14th day after modeling. Administering icariin to the mice means dispersing the required amount of icariin in corn oil to make a corn oil solution of icariin and administering it to the mice by gavage. The administered doses in each group were calculated based on the mass of icariin in the corn oil solution.

[0073] Time points for detecting bone cancer pain in mice:

[0074] (1) The day of modeling (day 0), the 3rd day, 7th day (the 1st day of drug administration), 10th day, 14th day after modeling, the 15th day of modeling, that is, the 1st day after drug withdrawal; the 16th day of modeling, that is, the 2nd day after drug withdrawal; the 18th day of modeling, that is, the 4th day after drug withdrawal; the 21st day of modeling, that is, the 7th day after drug withdrawal.

[0075] In the preliminary experiment, first according to the reference doses provided by Party A (35 / 70 / 140 mg / kg), the dose - time curve (S curve) was fitted according to log2[concentration] to determine the EC50, and the concentrations of 0 / 25 / 50 / 100 / 200 / 400 mg / kg were selected; then the optimal effective dose (such as 100 / 200 / 400 mg / kg) was determined according to the EC50 obtained from the fitted curve, and the final dose selection needed to be determined according to the EC50).

[0076] (2) Time points for detecting acute effects after drug withdrawal: On the 14th day after modeling, i.e., 1 h, 3 h, 6 h, 12 h, and 24 h after the second administration on the last day of drug administration.

[0077] The detection method is the same as that in Example 1.

[0078] The mechanical pain sensitivity, cold pain sensitivity, and spontaneous pain curves of the mice in the tumor model group, blank group, and phosphate buffer group are shown in Figure 4 、 Figure 5 and Figure 6 respectively.

[0079] It can be seen from Figure 4 that for the mice with a drug dosage of 200 mg / kg and 400 mg / kg, during the drug administration period from the 7th day to the 14th day, the paw withdrawal weight threshold was significantly enhanced compared to the control group mice. And after the drug administration ended, i.e., after the 14th day, although the paw withdrawal weight threshold of the mice was lower than that during the drug administration period, it was still much higher than that of the control group mice with a drug dosage of 0 mg / kg. By comparing groups 2, 3, 4, and 5, it can be seen that as the drug dosage increased from 50 mg / kg to 200 mg / kg, the paw withdrawal weight threshold of the mice continuously increased. After stopping the drug administration on the 14th day, the paw withdrawal weight threshold of each drug dosage group of mice decreased compared to that during the drug administration period. However, within the range of 50 mg / kg - 200 mg / kg of the drug dosage, as the previous drug dosage increased, the paw withdrawal weight also increased. The paw withdrawal weight threshold of the mice with a drug dosage of 200 mg / kg was basically consistent with that of the mice with a drug dosage of 400 mg / kg throughout the detection process.

[0080] It can be seen by comparing Figure 5 with Figure 2 that from group 2 to group 5, the cold pain sensitivity of the mice was much higher than that of the non-drug-administered mice. Compared with the non-drug-administered group, the cold pain sensitivity of the control group, i.e., the mice given corn oil, was lower than that of the non-drug-administered mice. The cold pain sensitivity of the mice in groups 3 - 5 was very close, including during and after the drug administration period, and was greater than that of the mice in group 1.

[0081] Figure 6 Let

[0082] Figure 7 The paw withdrawal latency threshold indicating the end of drug administration. As can be seen from the figure, within 7 days after the end of drug administration, the paw withdrawal latency thresholds of mice with drug doses of 100 mg / kg, 200 mg / kg, and 400 mg / kg are very close, and are greater than the paw withdrawal latency threshold of mice with a drug dose of 50 mg / kg. The paw withdrawal latency threshold of mice with a drug dose of 50 mg / kg is greater than the paw withdrawal latency threshold of mice in the corn oil group.

[0083] Example 3

[0084] Combined use of icariin and morphine

[0085] The bone cancer animal model of Example 1 was divided into the following groups:

[0086] Group 1: Intragastric administration of corn oil (solvent) group, i.e., the control group;

[0087] Group 2: Intragastric administration of morphine at a dose of 10 mg / kg each time, i.e., experimental group A;

[0088] Group 3: Intragastric administration of icariin at a dose of 100 mg / kg each time, i.e., experimental group B;

[0089] Group 4: Intragastric administration of icariin at a dose of 100 mg / kg and morphine at a dose of 1 mg / kg each time, i.e., experimental group C;

[0090] Group 5: Intragastric administration of icariin at a dose of 100 mg / kg and morphine at a dose of 2 mg / kg each time, i.e., experimental group D;

[0091] Group 6: Intragastric administration of icariin at a dose of 100 mg / kg and morphine at a dose of 4 mg / kg each time, i.e., experimental group E;

[0092] Group 7: Intragastric administration of icariin at a dose of 100 mg / kg and morphine at a dose of 8 mg / kg each time, i.e., experimental group F;

[0093] The experimental method was the same as that in Example 2, except that in addition to detecting the mechanical hyperalgesia, cold hyperalgesia, and the number of spontaneous paw lifts within 10 minutes of the mice, the paw lift protection time within 10 minutes was also detected to detect the sensitivity of the mice.

[0094] Through Figure 8 、 Figure 9 、 Figure 10 and Figure 11From the control group curve, it can be seen that since the 7th to 14th day after modeling, the mechanical pain withdrawal threshold (PWT) of the left hind paw (tumor-bearing side) and the cold pain hypersensitivity withdrawal latency threshold (PWL) of the bone cancer pain mice have significantly decreased, and the spontaneous pain raising foot frequency and raising foot protection time have significantly increased, and persist. Icariin was given on the 7th to 14th day after modeling. Since the 10th to 14th day, icariin significantly increased the PWT and PWL of the left hind paw of bone cancer pain (BCP) mice, and reduced the spontaneous raising foot frequency and raising foot protection time. The effect continued until the 18th day after modeling (i.e., the 4th day after drug withdrawal) (p < 0.05). The combination of icariin at 1-fold EC50 dose (100 mg / kg) and morphine significantly prolonged the analgesic effect after drug withdrawal, and the analgesic effect after drug withdrawal continued until the 21st day after modeling (i.e., the 7th day after drug withdrawal) (p < 0.05). The combination of icariin at half-effective dose (100 mg / kg) and morphine could reduce the morphine dosage by up to 60% when the optimal analgesic effect of morphine was achieved. Conclusion: Icariin has a significant analgesic effect on BCP mice. When combined with morphine, it not only significantly prolongs the analgesic effect after drug withdrawal, but also can reduce the morphine dosage by up to 60%.

[0095] Example 4

[0096] Icariin soft capsules are drugs already on the market. The preparation components and preparation methods refer to Chinese Patent: 201310526018.0.

[0097] Compare the quality of life of subjects with advanced liver cancer treated with icariin soft capsules and cinobufacini as the first-line treatment

[0098] Compare the core quality of life scales of the two groups of trial populations, and the quality of life is studied based on pain.

[0099] Dosage and administration of icariin soft capsules: soft capsules; specification 100 mg / capsule; oral administration; twice a day, 6 capsules (6 × 100 mg / capsule) / time; each 4 weeks is a medication treatment cycle; treatment duration: continuous medication until disease progression or intolerance.

[0100] Dosage and administration of cinobufacini tablets for comparison: tablets; specification 0.3 g / tablet; oral administration; three times a day, 4 tablets (4 × 0.3 g / tablet) / time; each 4 weeks is a medication treatment cycle; treatment duration: continuous medication until disease progression or intolerance. Pain index of patients before taking the medicine:

[0101] The enriched population uses a scoring method to evaluate the pain value of the subjects for scoring. The scoring level is divided into 4 levels. 1 point = none, 2 point = a little, 3 point = quite, 4 point = very. The higher the symptom dimension score, the worse the quality of life.

[0102] We made an evaluation for the two groups of patients taking icariin soft capsules and cinobufacini once when they were enrolled, and the scoring results are shown in Table 1. When both groups of patients had taken the medicine for 18 weeks on the last day, another evaluation was made, and the scoring results are shown in Table 2. In the middle of the 18-week trial, one patient dropped out from each group.

[0103] Table 1 Pain scoring of patients taking icariin soft capsules

[0104]

[0105] Table 2 Pain scoring of patients taking icariin soft capsules after 18 weeks of medication

[0106]

[0107] By comparing Table 1 and Table 2, it can be clearly seen that icariin soft capsules can relieve the pain of liver cancer patients.

Claims

1. Use of icariin in the preparation of a medicament for treating and / or relieving pain.

2. The use according to claim 1, wherein the pain comprises tumor-related pain.

3. The use according to claim 2, wherein the tumor comprises a solid tumor.

4. The use according to claim 2, wherein the tumor-related pain comprises pain associated with bone cancer.

5. The use according to claim 4, wherein the pain associated with bone cancer includes pain associated with primary bone cancer and secondary bone cancer.

6. The use according to claim 5, wherein the primary bone cancer includes chondrosarcoma, osteoblastic tumor and osteofibroma.

7. The use according to claim 5, wherein the secondary bone cancer comprises secondary bone cancer associated with one or more of breast cancer, lung cancer, prostate cancer, thyroid cancer, liver cancer, pancreatic cancer and colorectal cancer bone metastasis.

8. The use according to claim 3, wherein the solid tumor comprises one or more of breast cancer, lung cancer, prostate cancer, thyroid cancer, liver cancer, pancreatic cancer and colorectal cancer.

9. The use according to claim 1, wherein the medicine is an oral preparation.

10. The use according to claim 9, wherein the oral preparation is one or more of capsules, tablets, solutions, syrups, emulsions, suspensions and granules.

Images