WO2022121884A1 - 伊维菌素及其类似物在皮肤相关疾病治疗中的应用 - Google Patents

伊维菌素及其类似物在皮肤相关疾病治疗中的应用 Download PDF

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WO2022121884A1
WO2022121884A1 PCT/CN2021/136027 CN2021136027W WO2022121884A1 WO 2022121884 A1 WO2022121884 A1 WO 2022121884A1 CN 2021136027 W CN2021136027 W CN 2021136027W WO 2022121884 A1 WO2022121884 A1 WO 2022121884A1
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ivermectin
fluorouracil
skin
pharmaceutical composition
administration
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French (fr)
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丁佳隆
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广州海普丁生物科技有限公司
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C19/00—Cheese; Cheese preparations; Making thereof
    • A23C19/097—Preservation
    • A23C19/10—Addition of preservatives
    • A23C19/11—Addition of preservatives of antibiotics or bacteriocins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the invention relates to the technical field of biomedicine, in particular to an anti-tumor combined drug composition and application thereof.
  • Skin cancer is a malignant tumor that occurs in the skin, and its attack rate is the most common in Caucasians, followed by yellows, and the lowest in blacks. The most common age group is adults.
  • Skin cancer is mainly divided into malignant melanoma (MM) and non-melanoma skin cancer (NMSC).
  • non-melanoma skin cancer mainly includes basal cell carcinoma (Basal cell carcinoma, BCC) and squamous cell carcinoma (Squamous cell carcinoma, SCC).
  • Basal cell carcinoma Basal cell carcinoma
  • SCC squamous cell carcinoma
  • Most skin cancers occur on the face, neck, ears, forearms, and backs of the hands, and can occur from prolonged exposure to large amounts of sunlight, chronic chemical exposure, or prolonged exposure to radiation exposure, etc.
  • BCC basal cell carcinoma
  • SCC squamous cell carcinoma
  • the preferred treatment for skin cancer in modern medicine is surgery.
  • Early, small, well-differentiated skin cancers can be treated with physical therapy, including cryotherapy, electrotherapy, and laser therapy.
  • Radiotherapy has a cure rate of 95% for early clinical skin cancer, especially basal cell carcinoma, and is suitable for patients with surgical contraindications or difficult surgical resection.
  • Smaller skin cancers can achieve good results with the above treatments.
  • it will not only affect the patient's appearance, but also significantly increase the difficulty of treatment.
  • drug therapy since most skin cancers are not sensitive to chemotherapy drugs, they can only be used for adjuvant therapy after surgery or palliative therapy for advanced skin cancer.
  • Fluorouracil also known as 5-Fu or 5-Fluorouracil (5-Fluorouracil)
  • Fluorouracil also known as 5-Fu or 5-Fluorouracil
  • 5-Fu or 5-Fluorouracil has a molecular formula of C 4 H 3 FN 2 O 2 and a molecular weight of 130.08. Its chemical structure is similar to that of uracil, and it is a white crystal or powder.
  • Fluorouracil is a cell cycle specific drug and is currently clinically used in addition to adjuvant chemotherapy and palliative treatment for gastrointestinal cancers such as breast cancer, liver cancer and pancreatic cancer, ovarian cancer and primary bronchial lung adenocarcinoma, as well as malignant mole and hydatidiform moles.
  • gastrointestinal cancers such as breast cancer, liver cancer and pancreatic cancer, ovarian cancer and primary bronchial lung adenocarcinoma, as well as malignant mole and hydatidiform moles.
  • choriocarcinoma, serous cavity cancerous effusion, bladder cancer, and head and neck malignant tumors it can also be used for intratumoral injection.
  • fluorouracil is widely used in clinic and has significant and exact curative effect, it still has many shortcomings.
  • fluorouracil due to poor lipid solubility of fluorouracil, it is not easy to enter tissues and cells, and its bioavailability is low. The toxic and side effects of fluorouracil are large, which can cause bone marrow suppression and hemorrhagic enteritis, and the elimination in vivo is nonlinear and individual differences are large. The need for continuous intravenous administration and poor patient compliance limit the use of fluorouracil.
  • fluorouracil is used locally in the treatment of various skin cancers including melanoma and various viral warts (including recurrent condyloma acuminatum), keratosis, skin cancer, psoriasis (psoriasis), keloids and other diseases
  • fluorouracil topical preparations and local reactions such as erythema, burning, ulcers, scabs and other obvious adverse reactions.
  • the technical means to be adopted in the present invention is to use fluorouracil combined with ivermectin for skin cancer treatment.
  • Ivermectin combined with fluorouracil can greatly enhance the killing ability of skin tumor cells, improve the treatment effect, shorten the treatment period, and reduce the occurrence of side effects.
  • the inventive discovery of the present invention is the preventive and/or therapeutic effects of ivermectin and its analogs on skin-related diseases. Especially its application as a topical preparation. And when it is combined with fluorouracil, it shows an unexpected synergistic effect. therefore,
  • One aspect of the present invention provides the use of ivermectin and its analogs in the preparation of medicaments for treating and/or preventing skin-related diseases.
  • the skin-related diseases are selected from skin tumors, white plaques, condyloma acuminatum, common warts, flat warts, psoriasis, xeroderma pigmentosum, actinic keratosis, Bowen's disease, Bowen's papules disease, vitiligo, preferably skin tumors.
  • Skin tumors include malignant melanoma, squamous cell carcinoma, basal cell carcinoma, malignant lymphoma, idiopathic hemorrhagic sarcoma (Kaposi sarcoma), sweat gland carcinoma, dermatofibrosarcoma protuberans, angiosarcoma, and chest wall metastases from breast cancer .
  • the skin tumor is selected from skin cancer, preferably chest wall metastases of breast cancer, melanoma, basal cell carcinoma and squamous cell carcinoma, most preferably basal cell carcinoma and squamous cell carcinoma.
  • ivermectin and its analogs are selected from oral administration, parenteral administration or transdermal administration.
  • Parenteral administration is selected from intravenous injection, subcutaneous injection, intramuscular injection; oral administration and transdermal administration are preferred, and transdermal administration is more preferred.
  • the amount of ivermectin and its analogs is a prophylactically and/or therapeutically effective amount, for example a unit dose selected from 0.01-1000 mg, preferably 0.1-500 mg.
  • ivermectin and its analogs are preferred as the single active ingredient.
  • the dosage form is selected from tablets, capsules, pills, granules, solutions, suspensions, syrups, injections (including injections, injections sterile powders and concentrated solutions for injection), suppositories, inhalants or sprays, creams, ointments, lotions, gels, liniments, tinctures, patches or poultices; preferably creams, ointments, Lotions, gels, liniments. External preparations are preferred.
  • One aspect of the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising ivermectin and its analogues and fluorouracil, the pharmaceutical composition comprising the following compounds as active ingredients:
  • the ivermectin and its analogs are selected from avermectin, ivermectin, moxidectin, doramectin, ivermectin, moximectin, acaricide ivermectin, preferably ivermectin and moxidectin, more preferably ivermectin.
  • the pharmaceutical composition of the present invention wherein the weight ratio of ivermectin and its analogs to fluorouracil is selected from 0.001:1-1000:1, preferably 0.01:1-100:1, most preferably 0.05:1-50: 1.
  • the amount of ivermectin and its analogs is a prophylactically and/or therapeutically effective amount, for example, the unit dose is selected from 0.01-1000 mg, preferably 0.1-500 mg.
  • the amount of fluorouracil is a prophylactically and/or therapeutically effective amount, for example, the unit dosage range is selected from 0.01-1000 mg, preferably 0.1-500 mg.
  • composition of the present invention optionally comprising one or more pharmaceutically acceptable carriers and/or excipients, can be made into any pharmaceutically acceptable dosage form.
  • the dosage form is selected from tablets, capsules, pills, granules, solutions, suspensions, syrups, injections (including injections, sterile powders for injection and concentrated solutions for injection) , suppository, inhalation or spray, cream, ointment, lotion, gel, liniment, tincture, patch or poultice; preferably cream, ointment, lotion, gel, liniment .
  • the pharmaceutical composition of the present invention optionally further comprises one or more other therapeutic agents, and the other therapeutic agents are preferably therapeutic agents for skin-related diseases, such as skin tumors or skin cancers.
  • the pharmaceutical composition of the present invention is used for preparing a medicine for treating/or preventing skin-related diseases.
  • Another aspect of the present invention provides the use of ivermectin and its analogs in combination with fluorouracil in the preparation of a medicament for treating and/or preventing skin-related diseases.
  • the ivermectin and its analogs are selected from the group consisting of avermectin, ivermectin, moxidectin, doramectin, ivermectin, moxicin, and acaricide , preferably ivermectin and moxidectin, more preferably ivermectin.
  • the skin-related diseases are selected from skin tumors, white plaques, condyloma acuminatum, common warts, flat warts, psoriasis, xeroderma pigmentosum, actinic keratosis, Bowen's disease, Bowen's papules disease, vitiligo, preferably skin tumors.
  • Skin tumors include malignant melanoma, squamous cell carcinoma, basal cell carcinoma, malignant lymphoma, idiopathic hemorrhagic sarcoma (Kaposi sarcoma), sweat gland carcinoma, dermatofibrosarcoma protuberans, angiosarcoma, and chest wall metastases from breast cancer .
  • the skin tumor is selected from skin cancer, preferably chest wall metastases of breast cancer, melanoma, basal cell carcinoma and squamous cell carcinoma, most preferably basal cell carcinoma and squamous cell carcinoma.
  • ivermectin and its analogs and fluorouracil are independently selected from oral administration, parenteral administration or transdermal administration.
  • Parenteral administration is selected from intravenous injection, subcutaneous injection, intramuscular injection; oral administration and transdermal administration are preferred, and transdermal administration is more preferred.
  • ivermectin and its analogs and fluorouracil can be administered simultaneously, or sequentially (administer ivermectin and its analogs first, then fluorouracil, or first fluorouracil or ivermectin and its analogs).
  • sequential administration there may be a certain time interval or no time interval between the two administrations as required.
  • prophylactically and/or therapeutically effective amounts of ivermectin and its analogs and fluorouracil are sequentially administered to a subject or patient in need, or any pharmaceutical composition comprising both is administered.
  • ivermectin and its analogs and fluorouracil can be administered to a patient simultaneously or sequentially.
  • the method of the present invention is characterized in that the administration frequency of ivermectin and its analogs and fluorouracil can be once a day, or multiple times a day, or once a day, respectively.
  • Ivermectins and analogs thereof described in the present invention include but are not limited to Avermectin, Ivermectin, Moxidectin, Dormectin ), Eprinomectin, Moxidectin and Milbemycin, etc., preferably ivermectin and Moxidectin, more preferably ivermectin.
  • the ivermectin and its analogs of the present invention include the form of pharmaceutically acceptable salts thereof, and the fluorouracil includes the form of pharmaceutically acceptable salts thereof, and the salts may be the free base form of the compound with an organic acid and/or or any pharmaceutically acceptable salt formed from inorganic acids, organic bases and/or inorganic bases.
  • the skin-related diseases described in the present invention refer to any skin-related diseases, including but not limited to skin tumors, white patches, condyloma acuminatum, common warts, flat warts, psoriasis, xeroderma pigmentosum, and ray angle rheumatoid arthritis, Bowen's disease, Bowenoid papulosis, vitiligo, etc.; preferably skin-related neoplastic diseases, including benign tumors and malignant tumors (cancers), including but not limited to malignant melanoma, squamous cell carcinoma, basal cell carcinoma, malignant Lymphoma, idiopathic hemorrhagic sarcoma (Kaposi sarcoma), sweat gland carcinoma, dermatofibrosarcoma protuberance, angiosarcoma, and chest wall metastases of breast cancer, etc.; more preferably skin-related cancers, such as skin cancer; even more preferably, breast cancer Chest wall met
  • the "combination” described in the present invention is a mode of administration, which includes the simultaneous administration of prophylactically and/or therapeutically effective amounts of ivermectin and its analogs and fluorouracil, and also includes administration within a certain period of time. At least one prophylactically and/or therapeutically effective amount of said ivermectin and its analogs and at least one prophylactically and/or therapeutically effective amount of said fluorouracil, wherein both substances exhibit pharmacological effects. Said time period can be within one dosing cycle, preferably within 4 weeks, within 3 weeks, within 2 weeks, within 1 week, or within 24 hours.
  • the ivermectin and its analogs and the fluorouracil can be administered simultaneously or sequentially.
  • “Combination” includes the administration of prophylactically and/or therapeutically effective amounts of said ivermectin and its analogs and said fluorouracil, optionally by the same route of administration or by different routes of administration.
  • an "effective amount" of the present invention still refers to an amount of a drug effective to prevent and/or treat a disease or disorder in a mammal.
  • a therapeutically effective amount of the drug reduces the number of cancer cells, reduces the size of the tumor, inhibits (ie, to some extent slows and preferably prevents) the infiltration of cancer cells into surrounding organs, and/or reduces to some extent a one or more symptoms associated with the disorder.
  • ivermectin and its analogs with fluorouracil shows a significant synergistic therapeutic effect and enhances the antitumor activity. Compared with fluorouracil alone, the medication is safer, the treatment effect is better, and the patient's medication compliance is better.
  • Example 1 uses The chemiluminescence cell viability detection method (ie CTG method) was used to evaluate the growth inhibitory effect of ivermectin and fluorouracil combined administration and individual administration on human skin cancer cell lines.
  • CTG method chemiluminescence cell viability detection method
  • Cell lines skin cancer cell lines A431 (human skin basal cell carcinoma cells), SCL-1 (human skin squamous cell carcinoma cells), A375 (human skin melanoma cell carcinoma cells). All were purchased from the Cell Bank of the Chinese Academy of Sciences.
  • ivermectin and fluorouracil were purchased from Sigma, USA.
  • Cells in logarithmic growth phase include human skin cancer cell lines A431 (human skin basal cell carcinoma cells), SCL-1 (human skin squamous cell carcinoma cells), A375 (human skin melanoma cell carcinoma cells), at appropriate concentrations seeded in 96-well plates. After culturing the cells for 24 hours, the original medium was aspirated, and the test compound was added to the cell line. The cells were cultured for 3 days at 37°C, 5% CO2 and 95% humidity. CTG was added to read the plate on the Envision instrument, and the experiment was analyzed. data.
  • the experiment was divided into blank control group, ivermectin treatment group, fluorouracil treatment group and combined drug treatment group.
  • Final test concentrations 100 ⁇ M, 31.6 ⁇ M, 10 ⁇ M, 3.17 ⁇ M, 1 ⁇ M, 317nM, 100nM, 31.8nM, 10.1nM for ivermectin treatment group; 100 ⁇ M, 31.6 ⁇ M, 10 ⁇ M, 3.17 ⁇ M, 1 ⁇ M, 317nM for fluorouracil treatment group , 100nM, 31.8nM, 10.1nM; in the combined treatment group, the concentration of ivermectin was fixed at 3 ⁇ M (the cell growth inhibition rate was about 20-40%), and the concentration of fluorouracil was 100 ⁇ M, 31.6 ⁇ M, 10 ⁇ M, 3.17 ⁇ M, 1 ⁇ M , 317nM, 100nM, 31.8nM, 10.1nM.
  • Each administration group had inhibitory effect on the growth of human skin cancer cell lines A431, SCL-1 and A375. in:
  • the IC 50 values of the fluorouracil treatment group for inhibiting the growth of human skin cancer cell lines A431, SCL-1 and A375 were 8.2 ⁇ M, 6.2 ⁇ M, and 7.9 ⁇ M, respectively;
  • the IC 50 values of the ivermectin treatment group for inhibiting the growth of human skin cancer cell lines A431, SCL-1 and A375 were 5.3 ⁇ M, 4.2 ⁇ M and 4.9 ⁇ M, respectively;
  • the IC 50 values of fluorouracil inhibiting the growth of human skin cancer cell lines A431, SCL-1 and A375 in the combined treatment group were 0.4 ⁇ M, 0.2 ⁇ M and 0.2 ⁇ M, respectively.
  • Example 2 Inhibitory effect of combined administration of ivermectin and fluorouracil on in situ skin tumors in mice
  • SPF grade ICR mice male, 6 weeks old, weighing 16-18 g, were purchased from Qinglongshan Experimental Animal Breeding Center. Raised in a single cage, provided with sufficient feed and drinking water, maintained a circadian rhythm of 12h light and 12h darkness, temperature 25 ⁇ 2°C, humidity 50%-70%, and the experimental process followed animal ethics.
  • Phorbol ester (TPA) Sigma, USA.
  • Acetone Sinopharm Group Chemical Reagent Co., Ltd.
  • Ivermectin cream (specification 1%) and fluorouracil ointment (specification 5%) are both commercially available, blank ivermectin cream and blank fluorouracil ointment are prepared according to the drug instructions by selecting excipients, without the active ingredient ivermectin Cream with fluorouracil.
  • mice which are mice suitable for experiments.
  • mice were randomly divided into six groups and numbered. Dosing begins at this point.
  • Control group 1 The same amount of blank fluorouracil ointment was applied to the skin, 4 times a week, about once every 40 hours;
  • Control group 2 The same amount of blank ivermectin cream was applied to the skin, 4 times a week, about once every 40 hours;
  • Control group 3 Apply the same amount of blank fluorouracil ointment to the skin, and after absorption, apply the blank ivermectin cream to the skin, 4 times a week, about once every 40 hours;
  • Fluorouracil group apply fluorouracil ointment to the skin, 4 times a week, about once every 40 hours, at a dose of 100 mg/kg;
  • Ivermectin group apply ivermectin cream to the skin, 4 times a week, about once every 40 hours, at a dose of 100 mg/kg.
  • the combined administration group of ivermectin and fluorouracil apply fluorouracil ointment to the skin, and after absorption, apply ivermectin cream to the skin, 4 times a week, about once every 40 hours.
  • the dose of bacteriocin was 50 mg/kg and the dose of fluorouracil was 50 mg/kg.
  • the skin cancer tumor volume of each administration group decreased compared with the control group.
  • the ivermectin group was basically the same as the fluorouracil group.
  • the combined administration of ivermectin and fluorouracil had significantly higher inhibitory effects on in situ skin tumors in mice than the fluorouracil group and the ivermectin group; 100mg/kg, the dose of fluorouracil group was 100mg/kg; the dose of ivermectin in the combination group was 50mg/kg, the dose of fluorouracil was 50mg/kg, and the total dose was 100mg/kg), the combined tumor volume was 20% (90.9mm3/450.2mm3*100 ⁇ 20%) and 24% (90.9mm3/380.7mm3*100 ⁇ 24%) of fluorouracil group and ivermectin group had obvious synergistic effect.
  • Example 3 Inhibitory effects of combined administration of ivermectin and fluorouracil and individual administration of each on mouse melanoma.
  • Ivermectin cream (specification 1%) and fluorouracil ointment (specification 5%) are both commercially available, blank ivermectin cream and blank fluorouracil ointment are prepared according to the drug instructions by selecting excipients, without the active ingredient ivermectin Cream with fluorouracil.
  • B16-F10 cells were provided by the Shanghai Cell Bank of the Chinese Academy of Sciences, and were cultured in a DMEM-based incubator at 37°C, 5% CO2, and saturated humidity.
  • Preparation of B16-F10 cell suspension in the ultra-clean workbench according to the aseptic operation rules, 4-5 days before the experiment, the cells are recovered and passaged in T75 cell culture flasks, the number of cells in each culture flask is about 0.8 ⁇ 107 -1.0 ⁇ 107 cells; when the cells grow to 80%, discard the culture medium, add 4mL of PBS to wash twice, aspirate the PBS, and routinely digest; transfer to a 15ml centrifuge tube, centrifuge at 800rpm/3min, aspirate and discard the supernatant, Add 1 mL of 4°C pre-chilled PBS to prepare a cell suspension of 1 ⁇ 107 cells/mL, which is kept in ice for later use.
  • the C57BL/6 subcutaneous tumor-bearing mouse model was established: one day before the experiment, the skin of the right lower limb of the mouse was prepared, the day of the experiment was recorded as day 0, the weight was recorded in the clean table, and the mice were anesthetized by inhalation of isoflurane. Grab and pinch the skin on the back of the mouse's neck with the left hand to lift the mouse, expose the skin of the right armpit, and disinfect the grafted skin with 75% alcohol; hold the needle with the right hand and pierce the right flank subcutaneously of the mouse horizontally. Swing horizontally to prevent penetration into the muscle layer; inject 0.1 mL of cell suspension, withdraw the needle slowly, and then press the needle hole with an alcohol cotton ball to prevent leakage; continue to feed and observe the tumor formation. Dosing started on day 6.
  • Control group 1 Apply the same amount of blank fluorouracil ointment to the skin, 4 times a week, about once every 40 hours;
  • Control group 2 an equal amount of blank ivermectin cream is applied to the skin, 4 times a week, about once every 40 hours;
  • Control group 3 Apply the same amount of blank fluorouracil ointment to the skin, and after absorption, apply the blank ivermectin cream to the skin, 4 times a week, about once every 40 hours;
  • Fluorouracil group apply fluorouracil ointment to the skin, 4 times a week, about once every 40 hours, at a dose of 100 mg/kg;
  • Ivermectin group apply ivermectin cream to the skin, 4 times a week, about once every 40 hours, at a dose of 100 mg/kg.
  • the combined administration group of ivermectin and fluorouracil apply fluorouracil ointment to the skin, and after absorption, apply ivermectin cream to the skin, 4 times a week, about once every 40 hours.
  • the dose of bacteriocin was 50 mg/kg and the dose of fluorouracil was 50 mg/kg.
  • T/C (%) (average tumor growth volume of treatment group/average tumor growth volume of control group) ⁇ 100%.
  • the tumor observation test will end 3 weeks after administration (Day 21), the mice will be sacrificed by cervical dislocation, and the tumor volume will be measured.
  • the results showed that fluorouracil, ivermectin and combined administration could inhibit the growth of melanoma. specifically:
  • control group 1, control group 2, and control group 3 were basically the same.
  • the tumor volume was 71.8% of that in the control group 1, with a significant difference (P ⁇ 0.01).
  • the tumor volume was only 55.7% of that in the control group 2, with a significant difference (P ⁇ 0.01).
  • the tumor volume was 28.2% of the control group 3, with a significant difference (P ⁇ 0.01).
  • the skin cancer tumor volume of each administration group decreased, and the treatment effect of the ivermectin group was slightly better than that of the fluorouracil group.
  • the combined administration of ivermectin and fluorouracil had a significantly higher inhibitory effect on the transplanted skin tumors in mice than the fluorouracil group and the ivermectin group; /kg, the dose of fluorouracil group was 100 mg/kg; the dose of ivermectin in the combined administration group was 50 mg/kg, the dose of fluorouracil was 50 mg/kg, and the total dose was 100 mg/kg), and the combined tumor volume was 50 mg/kg of fluorouracil, respectively. 40% (324.1mm3/804.1mm3*100 ⁇ 40%) and 53% (324.1mm3/607.2mm3*100 ⁇ 53%) of the ivermectin group and the ivermectin group, with obvious synergistic effect.

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Abstract

提供一种伊维菌素及其类似物在预防和／或治疗皮肤相关疾病的应用。尤其是提供一种伊维菌素及其类似物和氟尿嘧啶联合在制备预防和／或治疗皮肤相关疾病药物中的用途。还提供了包含伊维菌素及其类似物和氟尿嘧啶的药物组合物及其用途。

Description

伊维菌素及其类似物在皮肤相关疾病治疗中的应用 技术领域
本发明涉及生物医药技术领域，尤其涉及一种抗肿瘤的联合用药组合物及其应用。
背景技术
皮肤癌是发生于皮肤的恶性肿瘤，其罹患率以白种人最常见，黄种人次之，黑种人最低，好发的年龄层以成年人为主。皮肤癌在临床上主要分为恶性黑色素瘤(Malignant melanoma，MM)和非黑色素瘤性皮肤癌(Non-melanoma skin cancer，NMSC)。其中，非黑色素瘤性皮肤癌主要包括基底细胞癌(Basal cell carcinoma，BCC)和鳞状细胞癌(Squamous cell carcinoma，SCC)。大部分的皮肤癌都发生在脸部、颈部、耳朵、前臂和手背等部位，其发生的原因包括长期大量的曝露于日光下、慢性化学药剂的接触，或在具有放射线的环境中长时间曝露等。随着户外休闲活动的增加以及大气臭氧层被破坏，全球皮肤癌的发生率及死亡率呈现逐渐增加的趋势，造成人体健康上的重大问题。
现代医学治疗皮肤癌的首选治疗方法为外科手术。早期、较小的、分化良好的皮肤癌，可以使用物理手段治疗，包括冷冻、电疗及激光治疗。放疗对临床早期皮肤癌，特别是基底细胞癌的治愈率可达95％，适合有手术禁忌症或手术切除有困难的患者。面积较小的皮肤癌采用上述治疗方式即可达到不错的治疗效果。但对于分布广或多发性的皮肤癌，不仅对患者会造成外观上的影响，治疗难度也显著提高。在药物治疗方面，由于大部分皮肤癌对化疗药物不敏感，只能用于手术后的辅助治疗或晚期皮肤癌的姑息治疗。
目前传统的治疗药物包括氟尿嘧啶(Fluorouracil)等。氟尿嘧啶又称5-Fu或5-氟尿嘧啶(5-Fluorouracil)，分子式为C 4H 3FN 2O 2，分子量为130.08，其化学结构与尿嘧啶类似，为白色结晶或粉末。氟尿嘧啶是细胞周期特异性药物，目前在临床上除用于乳腺癌、肝癌和胰腺癌等消化道癌肿、卵巢癌和原发性支气管肺腺癌的辅助化疗和姑息治疗，以及恶性葡萄胎和绒毛膜上皮癌、浆膜腔癌性积液和膀胱癌、头颈部恶性肿瘤的治疗外，还可用于瘤内注射。但是，尽管氟尿嘧 啶在临床上运用广泛、疗效显著而确切，但其仍存在诸多缺点。例如，由于氟尿嘧啶脂溶性差，不易进入组织和细胞，生物利用度低。氟尿嘧啶的毒副作用大，可引起骨髓抑制及出血性肠炎，并且体内消除非线形性、个体差异大。需要静脉持续用药，病人顺应性差，从而限制了氟尿嘧啶的应用。另外，虽然氟尿嘧啶在局部应用治疗包括黑色素瘤在内的各类皮肤癌及各类病毒疣(包括复发性尖锐湿疣)、角化病、皮肤癌、银屑病(牛皮癣)、瘢痕疙瘩等疾病方面取得了较好的进展，但同样存在氟尿嘧啶外用制剂治疗效果差，局部反应如红斑、灼热、溃疡、结痂等明显不良反应。
因此，提高氟尿嘧啶在皮肤癌领域的治疗效果，并减少产生的副作用方面，仍然存在较大的未满足的临床需求。本发明拟采用的技术手段是采用氟尿嘧啶联合伊维菌素进行皮肤癌治疗。伊维菌素联合氟尿嘧啶，可以极大的增效对皮肤肿瘤细胞的杀伤能力，提高了治疗效果，缩短治疗周期，同时减少副作用发生。
发明内容
本发明创造性的发现，伊维菌素及其类似物对皮肤相关疾病的预防和/或治疗作用。尤其是其作为外用制剂的应用。并且其与氟尿嘧啶联合治疗时，更是体现了预料不到的协同增效效果。因此，
本发明的一个方面，提供伊维菌素及其类似物在制备治疗和/或预防皮肤相关疾病药物中的用途。
本发明的用途，皮肤相关疾病选自皮肤肿瘤、白色斑块、疗尖锐湿疣、寻常疣、扁平疣、银屑病、着色性干皮病、光线性角化、博温病、博温样丘疹病、白癜风，优选皮肤肿瘤。皮肤肿瘤包括恶性黑色素瘤、鳞状细胞癌、基底细胞癌、恶性淋巴瘤、特发性出血性肉瘤(Kaposi肉瘤)、汗腺癌、隆突性皮肤纤维肉瘤、血管肉瘤，以及乳腺癌的胸壁转移。皮肤肿瘤选自皮肤癌，优选乳腺癌的胸壁转移、黑色素瘤、基底细胞癌和鳞状细胞癌，最优选基底细胞癌和鳞状细胞癌。
本发明的用途，伊维菌素及其类似物选自经口给药、胃肠外给药或经皮给药。胃肠外给药选自静脉注射、皮下注射、肌肉注射；优选口服给药和经皮给药，更优选经皮给药。
如上所述用途，伊维菌素及其类似物的量为预防和/或治疗有效量，例如单 位剂量选自0.01-1000mg，优选0.1-500mg。
如上所述用途，优选伊维菌素及其类似物作为单一活性成分。
如上所述用途，制成药学上可接受的任一剂型，所述剂型选自片剂、胶囊剂、丸剂、颗粒剂、溶液剂、混悬剂、糖浆剂、注射剂(包括注射液、注射用无菌粉末与注射用浓溶液)、栓剂、吸入剂或喷雾剂、乳膏剂、软膏剂、洗剂、凝胶剂、搽剂、酊剂、贴膜剂或巴布剂；优选乳膏剂、软膏剂、洗剂、凝胶剂、搽剂。优选外用制剂。
本发明的一个方面，提供一种包含伊维菌素及其类似物和氟尿嘧啶的药物组合物，所述药物组合物包含如下所述化合物作为活性成分：
A)伊维菌素及其类似物。
B)氟尿嘧啶。
本发明的药物组合物，所述伊维菌素及其类似物选自阿维菌素、伊维菌素、莫西菌素、多拉菌素、伊普菌素、摩西菌素、杀螨菌素，优选伊维菌素与莫西菌素，更优选伊维菌素。
本发明的药物组合物，其中所述伊维菌素及其类似物与氟尿嘧啶的重量比选自0.001:1-1000:1，优选0.01:1-100:1，最优选0.05:1-50:1。
本发明的药物组合物，其中伊维菌素及其类似物的量为预防和/或治疗有效量，例如单位剂量选自0.01-1000mg，优选0.1-500mg。
本发明的药物组合物，其中氟尿嘧啶的量为预防和/或治疗有效量，例如单位剂量范围选自0.01-1000mg，优选0.1-500mg。
本发明的药物组合物，任选包含一种或多种药学上可接受的载体和/或赋型剂，制成药学上可接受的任一剂型。
本发明的药物组合物，所述剂型选自片剂、胶囊剂、丸剂、颗粒剂、溶液剂、混悬剂、糖浆剂、注射剂(包括注射液、注射用无菌粉末与注射用浓溶液)、栓剂、吸入剂或喷雾剂、乳膏剂、软膏剂、洗剂、凝胶剂、搽剂、酊剂、贴膜剂或巴布剂；优选乳膏剂、软膏剂、洗剂、凝胶剂、搽剂。
本发明的药物组合物，任选进一步包含另外一种或多种其它治疗剂，所述其它治疗剂优选皮肤相关疾病治疗剂，例如皮肤肿瘤或皮肤癌。
本发明的药物组合物，其用于制备治疗/或预防皮肤相关疾病药物。
本发明的另一个方面，提供一种伊维菌素及其类似物与氟尿嘧啶联合在制备治疗和/或预防皮肤相关疾病药物中的用途。
本发明的用途，所述伊维菌素及其类似物选自阿维菌素、伊维菌素、莫西菌素、多拉菌素、伊普菌素、摩西菌素、杀螨菌素，优选伊维菌素与莫西菌素，更优选伊维菌素。
本发明的用途，皮肤相关疾病选自皮肤肿瘤、白色斑块、疗尖锐湿疣、寻常疣、扁平疣、银屑病、着色性干皮病、光线性角化、博温病、博温样丘疹病、白癜风，优选皮肤肿瘤。皮肤肿瘤包括恶性黑色素瘤、鳞状细胞癌、基底细胞癌、恶性淋巴瘤、特发性出血性肉瘤(Kaposi肉瘤)、汗腺癌、隆突性皮肤纤维肉瘤、血管肉瘤，以及乳腺癌的胸壁转移。皮肤肿瘤选自皮肤癌，优选乳腺癌的胸壁转移、黑色素瘤、基底细胞癌和鳞状细胞癌，最优选基底细胞癌和鳞状细胞癌。
本发明的用途，伊维菌素及其类似物和氟尿嘧啶分别独立选自经口给药、胃肠外给药或经皮给药。胃肠外给药选自静脉注射、皮下注射、肌肉注射；优选口服给药和经皮给药，更优选经皮给药。
本发明的用途，伊维菌素及其类似物和氟尿嘧啶可以同时给药，或顺次给药(先给予伊维菌素及其类似物，再给予氟尿嘧啶，或先氟尿嘧啶或伊维菌素及其类似物)。顺次给药时，两次给药间可以根据需要存在一定时间间隔或不存在时间间隔。
本发明的再一个方面，提供一种治疗和/或预防皮肤相关疾病的方法。
本发明的治疗方法，顺次给予需要的受试者或患者预防和/或治疗有效量的伊维菌素及其类似物和氟尿嘧啶，或给予包含两者的任意药物组合物。
本发明的治疗方法，伊维菌素及其类似物和氟尿嘧啶可以同时给予患者，或顺次给予患者。
本发明的方法，其特征在于，伊维菌素及其类似物和氟尿嘧啶的给药频次可以分别是一日一次，或一日多次，或多日一次。
术语说明：
本发明所述伊维菌素(Ivermectins)及其类似物，包括但不限于阿维菌素(Avermectin)，伊维菌素(Ivermectin)，莫西菌素(Moxidectin)，多拉菌素(Dornmectin)，伊普菌素(Eprinomectin)，摩西菌素(Moxidectin)和杀螨菌素 (Milbemycin)等，优选伊维菌素与莫西菌素(Moxidectin)，更优选伊维菌素。
本发明所述伊维菌素及其类似物包括其药学上接受的盐的形式，所述氟尿嘧啶包含其药学上可接受的盐的形式，所述盐可以是化合物游离碱形式与有机酸和/或无机酸、有机碱和/或无机碱形成的任意的药学上可接受到盐。
本发明所述皮肤相关疾病，指任意与皮肤相关的疾病，包括但不限于皮肤肿瘤、白色斑块、疗尖锐湿疣、寻常疣、扁平疣、银屑病、着色性干皮病、光线性角化、博温病、博温样丘疹病、白癜风等；优选与皮肤相关肿瘤疾病，包括良性肿瘤和恶性肿瘤(癌症)，包括但不限于恶性黑色素瘤、鳞状细胞癌、基底细胞癌、恶性淋巴瘤、特发性出血性肉瘤(Kaposi肉瘤)、汗腺癌、隆突性皮肤纤维肉瘤、血管肉瘤，以及乳腺癌的胸壁转移等；进一步优选皮肤相关癌症，例如皮肤癌；更进一步优选，乳腺癌的胸壁转移、黑色素瘤、基底细胞癌和鳞状细胞癌，最优选基底细胞癌和鳞状细胞癌。
本发明所述的“联合”是一种给药方式，包含同时给予预防和/或治疗有效量的伊维菌素及其类似物和氟尿嘧啶的给药方式外，还包含在一定时间期限内给予至少一种预防和/或治疗有效量的所述伊维菌素及其类似物和至少一种预防和/或治疗有效量的所述氟尿嘧啶，其中两种物质都显示药理学作用。所述的时间期限可以是一个给药周期内，优选4周内，3周内，2周内，1周内，或24小时以内。可以同时或顺次给予所述伊维菌素及其类似物似物和所述氟尿嘧啶。“联合”包括任选通过相同给药途径或不同给药途径给予预防和/或治疗有效量的所述伊维菌素及其类似物和所述氟尿嘧啶。
本发明“有效量”仍指在哺乳动物中有效预防和/或治疗疾病或病症的药物量。在癌症的情况中，治疗有效量的药物可减少癌细胞的数目，缩小肿瘤的尺寸，抑制(即一定程度的减缓和优选阻止)癌细胞浸润到周围器官中，和/或一定程度的减轻一种或多种与该病症有关的症状。
优益效果
伊维菌素及其类似物与氟尿嘧啶联合在预防和/或治疗癌症中，显示出明显的协同治疗效果，增强了抗肿瘤活性。相对单用氟尿嘧啶治疗，用药更安全、治疗效果更佳、患者用药依从性更好。
具体实施方式
以下通过实施例说明本发明的具体步骤，但不受实施例限制。
在本发明中所使用的术语，除非另有说明，一般具有本领域普通技术人员通常理解的含义。如下结合具体实例进一步详细描述本发明，以下实施例中未详细描述的过程和方法可以使用本领域中公知的常规方法。应理解，这些实施例只是为了举例说明本发明，而非以任何方式限制本发明的范围。
实施例1 采用
Figure PCTCN2021136027-appb-000001
化学发光细胞活率检测方法(即CTG方法)评价，伊维菌素与氟尿嘧啶联合给药与各自单独给药对人皮肤癌细胞株的生长抑制作用。
1、试验材料
细胞株：皮肤癌细胞株A431(人皮肤基底细胞癌细胞)、SCL-1(人皮肤鳞状细胞癌细胞)、A375(人皮黑色素瘤细胞癌细胞)。均购买自中国科学院细胞库。
待测化合物：伊维菌素、氟尿嘧啶，均购买于Sigma，美国。
2、试验方法
处于生长对数期的细胞包括人皮肤癌细胞株A431(人皮肤基底细胞癌细胞)、SCL-1(人皮肤鳞状细胞癌细胞)、A375(人皮黑色素瘤细胞癌细胞)，以适宜浓度种于96孔板中。细胞培养24h贴壁后吸去原来的培养基，向细胞系中加入待测化合物，在37℃、5％CO2和95％湿度条件下培养3天，加入CTG于Envision仪器上读板，分析实验数据。
试验分为空白对照组、伊维菌素处理组、氟尿嘧啶处理组、联合给药处理组。最终测试浓度：伊维菌素处理组为100μM，31.6μM，10μM，3.17μM，1μM，317nM，100nM，31.8nM，10.1nM；氟尿嘧啶处理组为100μM，31.6μM，10μM，3.17μM，1μM，317nM，100nM，31.8nM，10.1nM；联合给药处理组为伊维菌素浓度固定为3μM(细胞生长抑制率约为20-40％)，氟尿嘧啶浓度为100μM，31.6μM，10μM，3.17μM，1μM，317nM，100nM，31.8nM，10.1nM。
按如下方法加入CTG溶液检测细胞活性。
1)于室温平衡96孔板约30分钟。
2)每孔加入50μl CTG溶液。
3)用微孔板震荡器混匀2分钟使细胞裂解。
4)于室温放置20分钟稳定荧光信号。
5)用Envision2104读板仪测定荧光信号值。
6)应用GraphPad Prism软件，使用非线性回归模型绘制S型剂量-反应曲线并计算IC 50值。
3、试验结果
各给药组对人皮肤癌细胞株A431、SCL-1和A375的生长均具有抑制作用。其中：
氟尿嘧啶处理组抑制人皮肤癌细胞株A431、SCL-1和A375生长的IC 50值分别为：8.2μM、6.2μM、7.9μM；
伊维菌素处理组抑制人皮肤癌细胞株A431、SCL-1和A375生长的IC 50值分别为：5.3μM、4.2μM、4.9μM；
联合给药处理组抑制人皮肤癌细胞株A431、SCL-1和A375生长的氟尿嘧啶的IC 50值分别为：0.4μM、0.2μM、0.2μM。
由上述实施例表明：在含有伊维菌素3μM(细胞生长抑制率约为20-40％)的情况下，氟尿嘧啶对人皮肤癌细胞株A431、SCL-1和A375生长的抑制能力极大提升，氟尿嘧啶对A431、SCL-1和A375的IC 50值分别降低约20倍(8.2μM/0.4μM≈20)、30倍(6.2μM/0.2μM≈30)与40倍(7.9μM/0.2μM≈40)。
实施例2 伊维菌素与氟尿嘧啶联合给药与各自单独给药对小鼠皮肤原位皮肤肿瘤的抑制作用
1、试验材料
实验动物：
SPF级ICR小鼠，雄性，6周龄，体重16-18g，购自青龙山实验动物养殖中心。单笼饲养，提供充足的饲料和饮水，保持12h光照，12h黑暗的昼夜节律，温度25±2℃，湿度50％-70％，实验过程遵循动物伦理。
主要试剂：
7,12-二甲基苯蒽(DMBA)：Sigma，美国。
佛波酯(TPA)：Sigma，美国。
丙酮：国药集团化学试剂有限公司。
生理盐水：安徽双鹤药业有限责任公司。
水合氯醛：上海凌峰化学试剂有限公司。
伊维菌素乳膏(规格1％)与氟尿嘧啶软膏(规格5％)均为市售，空白伊维菌素乳膏与空白氟尿嘧啶软膏根据药品说明书选取辅料制备，不含活性成分伊维菌素乳膏与氟尿嘧啶。
2、化学诱导法建立小鼠皮肤肿瘤模型
7周周龄的C57BL/6小鼠(体重＝20g±2g)，用宠物剃毛刀剃小鼠背部毛皮(面积相似)，先使用300μg/mL的7,12-二甲基苯并(a)蒽(DMBA)涂抹200μL；随后每周两次涂抹佛波脂(TPA)(20μg/mL每次；200μL体积)涂抹20周。
试验开始7-10周后小鼠背部开始出现肿瘤块，从成模小鼠中选取适合进行下一步实验的个体，判断标准为：荷有1-3个体积介于100mm 3-200mm 3肿瘤的，为适宜进行实验的小鼠。
实验分组：将符合条件的小鼠随机分为六组，并进行编号。此时开始给药。
3、抗肿瘤药物治疗
对照组1：等量空白氟尿嘧啶软膏于皮肤涂抹，每周4次，约每40小时给药一次；
对照组2：等量空白伊维菌素乳膏于皮肤涂抹，每周4次，约每40小时给药一次；
对照组3：等量空白氟尿嘧啶软膏于皮肤涂抹，待吸收后，以空白伊维菌素乳膏于皮肤涂抹，均为每周4次，约每40小时给药一次；
氟尿嘧啶组：以氟尿嘧啶软膏于皮肤涂抹，每周4次，约每40小时给药一次，100mg/kg剂量；
伊维菌素组：以伊维菌素乳膏于皮肤涂抹，每周4次，约每40小时给药一次，100mg/kg剂量。
伊维菌素与氟尿嘧啶联合给药组：以氟尿嘧啶软膏于皮肤涂抹，待吸收后，以伊维菌素乳膏于皮肤涂抹，均为每周4次，约每40小时给药一次，伊维菌素剂量为50mg/kg，氟尿嘧啶剂量为50mg/kg。
分别测量每个肿瘤的长径(a)，短径(b)和高度(h)并计算肿瘤体积，肿瘤的体 积＝(a×b×h)×6/π。视情况对小鼠进行脱毛，持续给药和记录数据，肿瘤抑制实验持续60天。
表1 氟尿嘧啶对小鼠皮肤原位皮肤肿瘤的抑制作用
组别 平均肿瘤体积(mm 3)
对照组1 750.2±65.1N＝8
氟尿嘧啶组 450.2±35.8N＝8(P＜0.01)
表2 伊维菌素对小鼠皮肤原位皮肤肿瘤的抑制作用
组别 平均肿瘤体积(mm 3)
对照组2 730.2±75.2N＝8
伊维菌素组 380.7±31.1N＝8(P＜0.01)
表3 联合给药对小鼠皮肤原位皮肤肿瘤的抑制作用
组别 平均肿瘤体积(mm 3)
对照组3 790.2±55.1N＝8
联合给药组 90.9±8.7N＝8(P＜0.01)
表4 联合给药相对单药治疗的比较
组别 平均肿瘤体积(mm 3)
联合给药组 90.9±8.7N＝8(P＜0.01)
氟尿嘧啶组 450.2±35.8N＝8
伊维菌素组 380.7±31.1N＝8
4、试验结果
如表1-4所示，各给药组的皮肤癌肿瘤体积相较于对照组均有下降。伊维菌素组与氟尿嘧啶组基本相当。但伊维菌素与氟尿嘧啶联合给药对小鼠原位皮肤肿瘤的抑制作用显著高于氟尿嘧啶组与伊维菌素组；在总给药剂量均为100mg/kg(伊维菌素组剂量为100mg/kg，氟尿嘧啶组剂量为100mg/kg；联合给药组伊维菌素剂量为50mg/kg，氟尿嘧啶剂量为50mg/kg，总剂量为100mg/kg)情况下，联合给药肿瘤体积分别为氟尿嘧啶组与伊维菌素组20％(90.9mm3/450.2mm3*100≈20％)与24％(90.9mm3/380.7mm3*100≈24％)，具有明显的协同 作用。
实施例3：伊维菌素与氟尿嘧啶联合给药与各自单独给药对小鼠黑素瘤的抑制作用。
1、试验材料
试验药物：
伊维菌素乳膏(规格1％)与氟尿嘧啶软膏(规格5％)均为市售，空白伊维菌素乳膏与空白氟尿嘧啶软膏根据药品说明书选取辅料制备，不含活性成分伊维菌素乳膏与氟尿嘧啶。
实验动物：
雌性C57BL/6，6～8周龄，体重16-18g，购于上海斯莱克实验动物有限公司，SPF级环境(恒温25±2℃，恒湿40％-50％)喂养。
2、动物模型制备
黑色素瘤B16-F10细胞由中国科学院上海细胞库提供，使用DMEM完全培养基于37℃、5％CO2、饱和湿度的培养箱中培养。B16-F10细胞悬液制备：在超净工作台内按照无菌操作规则进行，实验前4-5d，将细胞复苏、传代于T75细胞培养瓶中，每个培养瓶细胞数大约含0.8×107-1.0×107个细胞；待细胞生长至80％时，弃培养液，加入4mL PBS清洗2遍，吸弃PBS，常规消化；转移至15ml离心管中，800rpm/3min离心，吸弃上清，加入1mL4℃预冷的PBS，制成1×107个/mL的细胞悬液，置于冰中备用。建立C57BL/6皮下荷瘤小鼠模型：实验前一天小鼠前右下肢备皮，实验当天记为day0，称重记录在超净台内，用异氟烷口鼻吸入，对小鼠进行麻醉用左手抓捏小鼠颈背部皮肤提起小鼠，暴露其右侧腋下皮肤，用75％酒精消毒移植处皮肤；右手持针水平刺入小鼠右侧胁肋部皮下，针头刺入后能水平摆动，以防止打入肌肉层；注入0.1mL细胞悬液，缓慢退针，然后用酒精棉球按压针孔，以防渗漏；继续饲养，观察成瘤情况，所有动物于造模后第6天开始给药。
3、干预措施
对照组1：等量空白氟尿嘧啶软膏于皮肤涂抹，每周4次，约每40小时一次；
对照组2：等量空白伊维菌素乳膏于皮肤涂抹，每周4次，约每40小时一 次；
对照组3：等量空白氟尿嘧啶软膏于皮肤涂抹，待吸收后，以空白伊维菌素乳膏于皮肤涂抹，均为每周4次，约每40小时一次；
氟尿嘧啶组：以氟尿嘧啶软膏于皮肤涂抹，每周4次，约每40小时给药一次，100mg/kg剂量；
伊维菌素组：以伊维菌素乳膏于皮肤涂抹，每周4次，约每40小时给药一次，100mg/kg剂量。
伊维菌素与氟尿嘧啶联合给药组：以氟尿嘧啶软膏于皮肤涂抹，待吸收后，以伊维菌素乳膏于皮肤涂抹，均为每周4次，约每40小时给药一次，伊维菌素剂量为50mg/kg，氟尿嘧啶剂量为50mg/kg。
肿瘤抑制的评价指标为相对肿瘤生长率：T/C(％)＝(治疗组的平均肿瘤增长体积/对照组的平均肿瘤增长体积)×100％。
4、试验结果
瘤体观察观试验将于给药后3周(Day 21)时结束，通过颈椎脱臼法处死小鼠，测量肿瘤体积。结果表明：氟尿嘧啶、伊维菌素和联合给药均可以抑制黑素瘤生长。具体地：
对照组1、对照组2、对照组3肿瘤体积基本相当。
氟尿嘧啶组干预2周末，肿瘤体积为对照组1的71.8％，具有显著性差异(P＜0.01)。
伊维菌素组干预2周末，肿瘤体积仅为对照组2的55.7％，具有显著性差异(P＜0.01)。
伊维菌素与氟尿嘧啶联合给药组干预2周，肿瘤体积为对照组3的28.2％，具有显著性差异(P＜0.01)。
表5 氟尿嘧啶对小鼠黑素瘤的抑制作用
组别 平均肿瘤体积(mm 3)
对照组1 1120.2±105.1N＝8
氟尿嘧啶组 804.1±75.8N＝8(P＜0.01)
抑制率T/C(％) 71.8％
表6 伊维菌素对小鼠黑素瘤的抑制作用
组别 平均肿瘤体积(mm 3)
对照组2 1090.2±113.2N＝8
伊维菌素组 607.2±51.5N＝8(P＜0.01)
抑制率T/C(％) 55.7％
表7 联合给药对小鼠黑素瘤的抑制作用
组别 平均肿瘤体积(mm 3)
对照组3 1149.2±142.2N＝8
联合给药组 324.1±29.8N＝8(P＜0.01)
抑制率T/C(％) 28.2％
表8 联合给药相对单药治疗的比较
组别 平均肿瘤体积(mm 3)
联合给药组 324.1±29.8N＝8(P＜0.01)
氟尿嘧啶组 804.1±75.8N＝8
伊维菌素组 607.2±51.5N＝8
结果表明：
各给药组的皮肤癌肿瘤体积相较于对照组均有下降，伊维菌素组治疗效果略优于氟尿嘧啶组。但伊维菌素与氟尿嘧啶联合给药对小鼠移植皮肤肿瘤的抑制作用显著高于氟尿嘧啶组与伊维菌素组；在总给药剂量均为100mg/kg(伊维菌素组剂量为100mg/kg，氟尿嘧啶组剂量为100mg/kg；联合给药组伊维菌素剂量为50mg/kg，氟尿嘧啶剂量为50mg/kg，总剂量为100mg/kg)情况下，联合给药肿瘤体积分别为氟尿嘧啶组与伊维菌素组的40％(324.1mm3/804.1mm3*100≈40％)与53％(324.1mm3/607.2mm3*100≈53％)，具有明显的协同作用。

Claims (23)

  1. 一种药物组合物，其特征在于，包含如下所述活性成分：
    A)伊维菌及其类似物，所述伊维菌素及其类似物选自阿维菌素、伊维菌素、莫西菌素、多拉菌素、伊普菌素、摩西菌素、杀螨菌素；优选伊维菌素与莫西菌素；更优选伊维菌素。
    B)氟尿嘧啶。
  2. 根据权利要求1所述药物组合物，其特征在于，伊维菌素及其类似物和氟尿嘧啶的量为预防和/或治疗有效量；优选地，
    伊维菌素及其类似物的单位剂量选自0.01-1000mg，优选0.1-500mg；
    氟尿嘧啶的单位剂量范围选自0.01-1000mg，优选0.1-500mg。
  3. 根据权利要求1-2任一项所述药物组合物，其特征在于，伊维菌素及其类似物与氟尿嘧啶的重量比选自0.001:1-1000:1；优选0.01:1-100:1，最优选0.05:1-50:1。
  4. 根据权利要求1-3任一项所述药物组合物，任选包含一种或多种药学上可接受的载体和/或赋型剂，制成药学上可接受的任一剂型。
  5. 根据权利要求4所述药物组合物，其特征在于，所述剂型选自片剂、胶囊剂、丸剂、颗粒剂、溶液剂、混悬剂、糖浆剂、注射剂(包括注射液、注射用无菌粉末与注射用浓溶液)、栓剂、吸入剂或喷雾剂、乳膏剂、软膏剂、洗剂、凝胶剂、搽剂、酊剂、贴膜剂或巴布剂；优选乳膏剂、软膏剂、洗剂、凝胶剂、搽剂。
  6. 根据权利要求1-5任一项所述药物组合物，其特征在于，所述药物组合物任选进一步包含另外一种或多种其它治疗剂，所述其它治疗剂优选皮肤相关疾病治疗剂，例如皮肤癌治疗剂。
  7. 权利要求1-6任一项所述药物组合物在制备治疗/或预防皮肤相关疾病药物中的用途。
  8. 伊维菌素及其类似物与氟尿嘧啶联合在制备治疗和/或预防皮肤相关疾病药物中的用途，所述伊维菌素及其类似物选自阿维菌素、伊维菌素、莫西菌素、多拉菌素、伊普菌素、摩西菌素、杀螨菌素，优选伊维菌素与莫西菌素，更优选伊维菌素。
  9. 根据权利要求8所述用途，其特征在于，所述皮肤相关疾病选自皮肤肿 瘤、白色斑块、疗尖锐湿疣、寻常疣、扁平疣、银屑病、着色性干皮病、光线性角化、博温病、博温样丘疹病、白癜风，优选皮肤肿瘤。
  10. 根据权利要求9所述用途，其特征在于，所述皮肤肿瘤包括恶性黑色素瘤、鳞状细胞癌、基底细胞癌、恶性淋巴瘤、特发性出血性肉瘤(Kaposi肉瘤)、汗腺癌、隆突性皮肤纤维肉瘤、血管肉瘤，以及乳腺癌的胸壁转移。
  11. 根据权利要求9所述用途，其中所述皮肤肿瘤选自皮肤癌，优选乳腺癌的胸壁转移、黑色素瘤、基底细胞癌和鳞状细胞癌，最优选基底细胞癌和鳞状细胞癌。
  12. 权利要求8-11任一项所述用途，其特征在于，伊维菌素及其类似物和氟尿嘧啶分别独立选自经口给药、胃肠外给药或经皮给药；优选口服给药和经皮给药，更优选经皮给药。
  13. 根据权利要求12所述用途，其特征在于，所述胃肠外给药选自静脉注射、皮下注射、肌肉注射。
  14. 根据权利要求8-13任一项所述用途，其特征在于，所述伊维菌素及其类似物和氟尿嘧啶类可以同时给药，或顺次给药(现给予伊维菌素及其类似物，再给予氟尿嘧啶，或先氟尿嘧啶或伊维菌素及其类似物)。
  15. 根据权利要求14所述用途，其特征在于，所述顺次给药时，两次给药间可以根据需要存在一定时间间隔或不存在时间间隔。
  16. 一种治疗和/或预防皮肤相关疾病的方法，其特征在于，同时或顺次给予需要的受试者或患者治疗有效量的伊维菌素及其类似物、氟尿嘧啶，或包含两者的药物组合物。
  17. 根据权利要求16所述的制备方法，所述药物组合物可以是权利要求1-6任一项所述的药物组合物。
  18. 根据权利要求16-17任一项所述的方法，其特征在于，伊维菌素及其类似物和氟尿嘧啶的给药频次可以分别是一日一次，或一日多次，或多日一次。
  19. 伊维菌素在制备治疗和/或预防皮肤相关疾病药物中的用途。
  20. 根据权利要求19所述用途，其特征在于，伊维菌素及其类似物的量为预防和/或治疗有效量，例如单位剂量选自0.01-1000mg，优选0.1-500mg。
  21. 根据权利要求19-20任一项所述用途，其特征在于，伊维菌素及其类似 物作为单一活性成分。
  22. 根据权利要求19-21任一项所述用途，制成药学上可接受的任一剂型。
  23. 权利要求22所述用途，其特征在于，所述剂型选自片剂、胶囊剂、丸剂、颗粒剂、溶液剂、混悬剂、糖浆剂、注射剂(包括注射液、注射用无菌粉末与注射用浓溶液)、栓剂、吸入剂或喷雾剂、乳膏剂、软膏剂、洗剂、凝胶剂、搽剂、酊剂、贴膜剂或巴布剂；优选乳膏剂、软膏剂、洗剂、凝胶剂、搽剂；优选外用制剂。
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101390828A (zh) * 2007-02-12 2009-03-25 济南康泉医药科技有限公司 同载抗代谢类药物和增效剂的抗癌组合物
CN101407775A (zh) * 2008-11-27 2009-04-15 中国科学院微生物研究所 一种制备阿维菌素的方法及其专用菌株
CN102517321A (zh) * 2011-12-28 2012-06-27 上海交通大学 阿维菌素的优化生产方法
CN102870790A (zh) * 2012-09-13 2013-01-16 青岛文创科技有限公司 伊维菌素和壳寡糖的增效农药组合物
BR102018009061A2 (pt) * 2018-05-04 2018-10-23 Biotec Biológica Indústria Farmacêutica Ltda. - E.P.P. composição farmacêutica compreendendo ivermectina em forma de creme dermatológico para o tratamento da rosacea e seu respectivo processo de obtenção

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101390828A (zh) * 2007-02-12 2009-03-25 济南康泉医药科技有限公司 同载抗代谢类药物和增效剂的抗癌组合物
CN101407775A (zh) * 2008-11-27 2009-04-15 中国科学院微生物研究所 一种制备阿维菌素的方法及其专用菌株
CN102517321A (zh) * 2011-12-28 2012-06-27 上海交通大学 阿维菌素的优化生产方法
CN102870790A (zh) * 2012-09-13 2013-01-16 青岛文创科技有限公司 伊维菌素和壳寡糖的增效农药组合物
BR102018009061A2 (pt) * 2018-05-04 2018-10-23 Biotec Biológica Indústria Farmacêutica Ltda. - E.P.P. composição farmacêutica compreendendo ivermectina em forma de creme dermatológico para o tratamento da rosacea e seu respectivo processo de obtenção

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
CHEN LIANG, BI SHUNING, WEI QIUREN, ZHAO ZHIJUN, WANG CHAOJIE, XIE SONGQIANG: "Ivermectin suppresses tumour growth and metastasis through degradation of PAK1 in oesophageal squamous cell carcinoma", JOURNAL OF CELLULAR AND MOLECULAR MEDICINE, UNIVERSITY PRESS CAROL DAVILA, BUCHAREST, RO, vol. 24, no. 9, 1 May 2020 (2020-05-01), RO , pages 5387 - 5401, XP055941178, ISSN: 1582-1838, DOI: 10.1111/jcmm.15195 *

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