Use of an extract from rabbit skin inflamed by vaccinia virus in the treatment of hematopoietic damage

Abstract

The present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in treatment. More specifically, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in treatment of hematopoietic system damage or pancytopenia induced by cancer therapy. Furthermore, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in treatment of leukopenia induced by cancer therapy. Furthermore, the extract derived from rabbit skin inflamed by vaccinia virus may be suitable for regeneration.

Description

[Technical Field]

[0001] The present invention relates to the field of medicine. Specifically, the present invention relates to a novel therapeutic use of an extract derived from rabbit skin inflamed by vaccinia virus. More specifically, the present invention relates to the use of said extract in the treatment of hematopoietic damage or pancytopenia induced by cancer therapy. [Background technology]

[0002] Although regular health checkups have enabled early diagnosis and detection of cancer, and the rate of successful surgical resection of primary cancer has steadily improved, various cancer treatments (including radiation therapy or chemotherapy) are still used to treat advanced or metastatic cancer.

[0003] Cancer treatment not only damages cancer cells, but also damages actively dividing normal cells, such as bone marrow hematopoietic cells. Side effects of anticancer drugs include hematopoietic dysfunction and bone marrow suppression, the main symptom of which is a decrease in white blood cells, with both red blood cells and platelets decreasing to varying degrees. When undergoing cancer treatment, patients' bodies may not produce enough white blood cells to resist invading bacteria and viruses, making them highly susceptible to life-threatening infections.

[0004] Under these circumstances, there is an urgent need for the development of drugs to alleviate or treat hematopoietic dysfunction and bone marrow suppression induced by cancer treatment.

[0005] It is known that nilestriol can be used to alleviate hematopoietic damage caused by anti-cancer radiation therapy (see Lirong Yi et.al, Chinese Journal of Radiological Medicine and Protection, June 2016, Vol. 36(6): 412-417). However, there is still a need to develop other drugs for cancer therapy, especially for the treatment of hematopoietic damage caused by anti-cancer drugs.

[0006] The "extract from rabbit skin inflamed by vaccinia virus" referred to herein refers to an active substance extracted from rabbit skin inflamed by inoculation with vaccinia virus, as described in Chinese Patent ZL98103220.6 (Chinese Patent No. CN1055249C), the entire contents of which are incorporated herein by reference. Such an extract from rabbit skin inflamed by vaccinia virus is commercially available under the trade name Lepalvir, manufactured by Weishi Pharmaceutical (Rugao) Co., Ltd. The extract from rabbit skin inflamed by vaccinia virus and its pharmacological effects are also described in WO2010 / 054531, the entire contents of which are incorporated herein by reference. However, until the present invention, it was unknown whether the extract was effective in treating hematopoietic damage or pancytopenia induced by cancer therapy. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Chinese Patent No. 1055249 [Patent Document 2] International Publication No. 2010 / 054531 [Non-patent literature]

[0008] [Non-Patent Document 1] Lirong Yi et.al, Chinese Journal of Radiological Medicine and Protection, 2016.6, Vol.36(6): 412-417 Summary of the Invention [Problem to be solved by the invention]

[0009] The object of the present invention includes providing a medicament for preventing, alleviating, or treating hematopoietic damage or pancytopenia induced by cancer therapy. More specifically, the object of the present invention includes providing a medicament for preventing, alleviating, or treating leukopenia induced by cancer therapy. [Means for solving the problem]

[0010] The technical problem of the present invention is solved by providing an extract, preferably a regenerating extract, derived from rabbit skin inflamed by vaccinia virus. [Effects of the Invention]

[0011] Generally speaking, the inventors have discovered that an extract derived from rabbit skin inflamed by vaccinia virus can effectively prevent, treat, or alleviate side effects induced by cancer therapy. Specifically, the extract can effectively prevent, treat, or alleviate hematopoietic damage or bone marrow suppression induced by cancer therapy. More specifically, the extract can effectively prevent, treat, or alleviate leukopenia induced by cancer therapy.

[0012] It is important to emphasize that leukopenia induced by cancer therapy may differ from that caused by other common medications. For example, antibiotics (e.g., phorbol esters) can induce platelet and leukocyte aggregation, and some anti-tuberculosis drugs (e.g., isoniazid) can induce antibodies in the body that bind to leukocytes. All of these can cause the destruction or reduction of leukocytes in the bloodstream. However, all of these processes affect existing and newly produced leukocytes, are relatively short-lived, and do not involve hematopoietic damage or bone marrow suppression. In contrast, cancer therapy not only kills cancer cells but also kills other actively proliferating cells, such as bone marrow hematopoietic cells. Therefore, leukopenia induced by anticancer drugs differs significantly from that caused by these drugs.

[0013] Furthermore, the inventors have discovered that an extract derived from rabbit skin inflamed by vaccinia virus can effectively restore or increase the body weight of patients undergoing cancer treatment, and the restoration or increase in body weight can be an indicator of the effectiveness of the extract in preventing or treating the disease or condition.

[0014] In one aspect, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in the manufacture of a medicament for preventing or treating hematopoietic system damage induced by cancer therapy in a patient. In one aspect, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in the prevention or treatment of hematopoietic system damage induced by cancer therapy. In one aspect, the present invention relates to a method for preventing or treating hematopoietic system damage induced by cancer therapy, the method comprising administering a therapeutically effective amount of an extract derived from rabbit skin inflamed by vaccinia virus to a patient in need thereof. In said aspect, the hematopoietic system damage comprises bone marrow suppression. In said aspect, the hematopoietic system damage or bone marrow suppression manifests as leukopenia.

[0015] In one embodiment, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in the manufacture of a medicament for preventing or treating myelosuppression induced by cancer therapy in a patient. In one embodiment, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in the prevention or treatment of myelosuppression induced by cancer therapy. In one embodiment, the present invention relates to a method for preventing or treating myelosuppression induced by cancer therapy, the method comprising administering a therapeutically effective amount of an extract derived from rabbit skin inflamed by vaccinia virus to a patient in need thereof. In said embodiment, the hematopoietic system damage or myelosuppression is manifested by leukopenia. In said embodiment, the myelosuppression is manifested by leukopenia.

[0016] In one embodiment, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in the manufacture of a medicament for restoring or improving hematopoietic function in a patient undergoing cancer treatment. In one embodiment, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in restoring or improving hematopoietic function in a patient undergoing cancer treatment. In one embodiment, the present invention relates to a method for restoring or improving hematopoietic function in a patient undergoing cancer treatment, the method comprising administering a therapeutically effective amount of an extract derived from rabbit skin inflamed by vaccinia virus to the patient undergoing cancer treatment. In the embodiment, the restoration or improvement of hematopoietic function comprises elimination or reduction of bone marrow suppression. In the embodiment, the restoration or improvement of hematopoietic function comprises an increase in white blood cell count.

[0017] In one aspect, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in the manufacture of a medicament for relieving or reducing bone marrow suppression in a patient undergoing cancer treatment. In one aspect, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus to relieving or reducing bone marrow suppression in a patient undergoing cancer treatment. In one aspect, the present invention relates to a method for relieving or reducing bone marrow suppression in a patient undergoing cancer treatment, the method comprising administering a therapeutically effective amount of an extract derived from rabbit skin inflamed by vaccinia virus to a patient undergoing cancer treatment. In the above aspect, the relieving or reducing bone marrow suppression comprises an increase in white blood cell count.

[0018] In one aspect, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in the manufacture of a medicament for preventing or treating pancytopenia induced by cancer therapy in a patient. In one aspect, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus to prevent or treat pancytopenia induced by cancer therapy in a patient. In one aspect, the present invention relates to a method for preventing or treating pancytopenia induced by cancer therapy in a patient, the method comprising administering to the patient a therapeutically effective amount of an extract derived from rabbit skin inflamed by vaccinia virus. In this aspect, the pancytopenia includes leukopenia.

[0019] In one aspect, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in the manufacture of a medicament for restoring or increasing total blood cells in a patient undergoing cancer treatment. In one aspect, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in the restoration or increase of total blood cells in a patient undergoing cancer treatment. In one aspect, the present invention relates to a method for restoring or increasing total blood cells in a patient undergoing cancer treatment, the method comprising administering to the patient a therapeutically effective amount of an extract derived from rabbit skin inflamed by vaccinia virus. In one aspect, the total blood cells are white blood cells.

[0020] In one embodiment, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in the manufacture of a medicament for preventing, alleviating, or treating weight loss induced by cancer therapy in a patient. In another embodiment, the present invention relates to the use of an extract derived from rabbit skin inflamed by vaccinia virus in the prevention, alleviating, or treating weight loss induced by cancer therapy in a patient. In another embodiment, the present invention relates to a method for alleviating or treating weight loss induced by cancer therapy in a patient, the method comprising administering to the patient a therapeutically effective amount of an extract derived from rabbit skin inflamed by vaccinia virus. In this embodiment, the patient is a patient undergoing cancer treatment. In this embodiment, the extract derived from rabbit skin inflamed by vaccinia virus is used to increase the weight of a patient undergoing cancer treatment. It should be emphasized that weight recovery or gain in a patient undergoing antibody therapy can be an indicator of recovery of hematopoietic function or alleviation of bone marrow suppression in the patient, and therefore may further indicate the therapeutic effect of the extract of the present invention.

[0021] In one aspect, the present invention relates to a combination pharmaceutical composition comprising an extract derived from rabbit skin inflamed by vaccinia virus and any pharmaceutically acceptable carrier, adjuvant, or excipient. In one aspect of the present invention, the pharmaceutically acceptable carrier, adjuvant, or excipient is used to formulate the pharmaceutical into an oral or injectable preparation. In one aspect, the extract derived from rabbit skin inflamed by vaccinia virus is formulated into an oral or injectable preparation, preferably an intramuscular or intravenous injection. In one aspect, the extract derived from rabbit skin inflamed by vaccinia virus is suitable for reinstatement.

[0022] In one embodiment of the present invention, the cancer treatment comprises radiation therapy or anticancer drug treatment. In one embodiment of the present invention, the cancer treatment comprises administering a cytotoxic drug to the patient. In one embodiment of the present invention, the anticancer drug comprises an agent that directly acts on DNA. In one embodiment of the present invention, the anticancer drug comprises an alkylating agent, more preferably a nitrogen mustard, such as cyclophosphamide. The dosage and administration method of cancer treatment can be selected by those skilled in the art based on the actual situation, and therefore are not conditions that limit the practice of the present invention.

[0023] In one embodiment of the present invention, the extract (preferably a reconstituted extract) derived from rabbit skin inflamed by vaccinia virus is administered to a patient, preferably a human, in an amount of about 0.01 to about 5 U / kg, preferably about 0.1 to about 2.5 U / kg, and more preferably about 0.15 to about 1.15 U / kg. For example, the extract derived from rabbit skin inflamed by vaccinia virus may be administered in an amount of about 0.01 U / kg, about 0.02 U / kg, about 0.05 U / kg, about 0.1 U / kg, about 0.15 U / kg, about 0.2 U / kg, about 0.25 U / kg, about 0.3 U / kg, about 0.35 U / kg, about 0.5 U / kg, about 0.6 U / kg, about 0.8 U / kg, about 1 U / kg, about 1.05 U / kg, or about 1.1 U / kg. The compound is administered to a patient, preferably a human, in an amount selected from the range of about 1.13 U / kg, about 1.14 U / kg, about 1.15 U / kg, about 1.17 U / kg, about 1.18 U / kg, about 1.2 U / kg, about 1.3 U / kg, about 1.4 U / kg, about 1.5 U / kg, about 1.8 U / kg, about 2 U / kg, about 2.5 U / kg, about 3.5 U / kg, about 4.5 U / kg, and ranges including these numbers as upper and lower limits. Those skilled in the art know that the dosage is human dose (U / kg or mg / kg) = mouse dose (U / kg or mg / kg) / 12.3, or human dose (U / kg or mg / kg) = mouse dose (U / kg or mg / kg) × 0.08. The doses described above may be effective amounts for treating the disease in a patient. In one embodiment, the extract from rabbit skin inflamed by vaccinia virus is administered by injection, for example, intramuscular or intravenous injection, at the doses described above.

[0024] In one embodiment of the present invention, the produced pharmaceutical preparation contains about 0.6 to about 600 U, preferably about 6 to about 300 U, and more preferably about 9 to about 70 U of an extract derived from rabbit skin inflamed by vaccinia virus. The pharmaceutical preparation is administered to a human, for example, an adult human. The average weight of an adult human is, for example, 60 kg. Therefore, the amount of extract from rabbit skin inflamed by vaccinia virus contained in the medicament prepared in the present invention is, for example, about 0.6 U, about 1.2 U, about 3 U, about 3.5 U, about 3.6 U, about 6 U, about 9 U, about 12 U, about 15 U, about 18 U, about 21 U, about 30 U, about 36 U, about 40 U, ​​about 60 U, about 63 U, about 66 U, about 67.8 U, about 68.4 U, about 69 U, about 69.5 U, about 70 U, about 70.2 U, about 70.8 U, about 72 U, about 78 U, about 84 U, about 90 U, about 108 U, about 120 U, about 150 U, about 210 U, about 270 U, and ranges with these upper or lower limits. In one embodiment, the medicament is prepared as an injection, for example, an intramuscular injection or an intravenous injection. In one embodiment, the medicament or injection is a fixed dose that cannot be divided. In one embodiment, the medicament or injection cannot be divided into smaller doses within 1, 2, 3, 4, 5, 6, or 7 days. In one embodiment, the medicament or injection is administered only once within 1, 2, 3, 4, 5, 6, or 7 days.

[0025] In one embodiment of the present invention, the extract derived from rabbit skin inflamed by vaccinia virus may be administered simultaneously with, separately from, or sequentially with cancer treatment. In one embodiment, the extract may be administered before or after cancer treatment. In one embodiment, the extract derived from rabbit skin inflamed by vaccinia virus is administered to the patient about 1 to 7 days, preferably 1 to 6 days, e.g., within 1, 2, 3, 4, or 5 days, after administration of cancer treatment to the patient. In one embodiment, the extract derived from rabbit skin inflamed by vaccinia virus is administered to the patient about 1 to 72 hours, preferably about 2 to 48 hours, more preferably about 3 to 24 hours, more preferably about 5 to 12 hours, more preferably about 6 to 8 hours, or more preferably about 6 hours after administration of cancer treatment to the patient. In one embodiment of the present invention, the extract derived from rabbit skin inflamed by vaccinia virus is administered to the patient about every 6 to 72 hours, preferably about 12 to 60 hours, more preferably about 24 to 48 hours, more preferably about 36 to 48 hours, or more preferably every 48 hours. In one embodiment of the present invention, a patient undergoing cancer treatment is administered an extract derived from rabbit skin inflamed by vaccinia virus continuously for at least about 24 months, at least about 12 months, at least about 6 months, at least about 2 months, at least about 1 month, at least about 3 weeks, at least about 2 weeks, at least about 10 days, at least about 7 days, at least about 5 days, or at least about 2 days.

[0026] In one embodiment, 1 to 12 days, preferably 2 to 8 days, and more preferably 4 to 7 days (e.g., 4, 5, 6, or 7 days) after administration, an extract derived from rabbit skin inflamed by vaccinia virus induces an increase in leukocytes in a patient that is greater than that observed in a patient not receiving the extract. In this embodiment, the increase in leukocytes observed in a patient receiving the extract is at least about 1-fold, at least about 1.01-fold, at least about 1.05-fold, at least about 1.1-fold, at least about 1.2-fold, at least about 1.3-fold, at least about 1.4-fold, at least about 1.5-fold, at least about 1.8-fold, at least about 2-fold, at least about 2.5-fold, at least about 5-fold, at least about 8-fold, at least about 10-fold, or at least about 20-fold, e.g., about 1.01 to about 2-fold, compared to the increase in leukocytes observed in a patient not receiving the extract.

[0027] In one embodiment of the present invention, the patient or animal subject may be a mammal, preferably a human. In this embodiment, the patient is a human patient undergoing cancer treatment. In another embodiment, the patient is a human patient suffering from hematopoietic damage or bone marrow suppression induced by cancer treatment. Furthermore, the leukopenia or leukopenia in the human patient is induced or caused by hematopoietic damage or bone marrow suppression.

[0028] As used herein, the terms "extract from rabbit skin inflamed by vaccinia virus" and "extract from rabbit skin inflamed by smallpox vaccine" can be used interchangeably and refer to a type of extract containing active substances extracted from rabbit skin inflamed by inoculation with vaccinia virus, for example, by processes such as leaching, purification, and repurification. This extract is usually a yellow or pale yellow liquid, but it can also be dried to form a solid. An injectable form of this type of extract from rabbit skin inflamed by vaccinia virus is commercially available under the trade name Lepalvir and produced by Weishi Pharmaceutical (Rugao) Co., Ltd. In one aspect, methods for preparing extracts or regenerating extracts from rabbit skin inflamed by vaccinia virus are described in Chinese Patent Publications CN1205233A, CN1613305A, and CN1493302A, PCT International Publication WO2004 / 060381, and European Patent Publication EP1557171, etc., the entire contents of which are incorporated herein by reference.

[0029] In one embodiment, an extract derived from rabbit skin inflamed by vaccinia virus or reconstituted therefrom can be prepared by a method comprising the following steps:

[0030] (1) collecting rabbit skin inflamed by inoculation with vaccinia virus, crushing the rabbit skin, and extracting the skin with an extraction solvent to obtain a solution A; (2) preparing solution A to be acidic and heating it to obtain solution B; (3) preparing solution B basic and heating it to obtain solution C; (4) adsorbing and filtering solution C under acidic conditions and eluting under basic conditions to obtain solution D; (5) neutralizing and heating solution D to obtain solution E; (6) concentrating solution E to obtain the extract; and (7) Optionally, mixing the extract with a pharmaceutically acceptable carrier, adjuvant, or excipient.

[0031] In one embodiment, in step (1), a vaccinia virus is inoculated into a domestic rabbit, the infected skin is collected, the skin is crushed, an aqueous phenol solution is added, and the skin is immersed at a temperature below about 12°C (e.g., about 0 to 10°C, preferably about 2 to 8°C, more preferably about 3 to 6°C, more preferably about 4°C) for at least about 12 hours (e.g., about 24 to 90 hours, preferably about 48 to 72 hours, more preferably about 70 or about 72 hours), centrifuged to obtain a supernatant, and filtered to obtain Solution A. The concentration of phenol in the aqueous phenol solution is about 1% to 10%, preferably about 2% to 5%, more preferably about 2% or about 3%.

[0032] In one embodiment, in step (2), solution A is acidified with an acid (e.g., hydrochloric acid) to, for example, about pH 4 to 6, more preferably about pH 4.5 to 5.5, more preferably about pH 5, and heated (e.g., at about 90 to 100°C, preferably about 95°C, continuously for at least about 10 minutes, for example, about 20 to 50 minutes, preferably about 30 to 40 minutes), optionally lowered to a temperature (e.g., below about 50°C, preferably below about 30°C), centrifuged to obtain a supernatant, and filtered to obtain solution B. Step (2) may be performed in a nitrogen environment.

[0033] In one embodiment, in step (3), solution B is adjusted to basicity (e.g., about pH 8 to 10, more preferably about pH 8.5 to 9.5, more preferably about pH 9 or about pH 9.2) using a base (e.g., sodium hydroxide), heated (e.g., at about 90 to 100°C, preferably about 95°C, continuously for at least 10 minutes, e.g., about 30 to 50 minutes, preferably about 30 to 40 minutes), optionally cooled (e.g., to a temperature below about 50°C, preferably below about 30°C), and filtered to obtain solution C. Step (3) may be performed in a nitrogen environment.

[0034] In one embodiment, in step (4), solution C is adjusted to acidic (e.g., about pH 3-6, more preferably about pH 4-5, more preferably about pH 4.5) using an acid (e.g., hydrochloric acid), and an adsorbent (e.g., activated carbon) is added thereto and immersed (e.g., for at least about 1 hour, preferably about 2-10 hours, more preferably about 4 hours, with continuous stirring). The solution is then removed to collect the adsorbent containing the active ingredient. The adsorbent is then added to an eluate (e.g., water), and the pH is adjusted to basic (e.g., about pH 9-12, preferably about pH 10 or 11) using a base (e.g., sodium hydroxide). The active ingredient is separated from the adsorbent (e.g., stirred for at least about 1 hour, preferably 2-10 hours, more preferably 4 hours, followed by filtration and washing the adsorbent with water), to obtain solution D. Step (4) may be performed under a nitrogen atmosphere.

[0035] In one embodiment, in step (5), solution D is neutralized to a weak acidity (e.g., about pH 5.5 to 6.6, preferably about pH 6) using an acid (e.g., hydrochloric acid) to obtain solution E. Preferably, step (5) may be performed under sterile conditions.

[0036] In one embodiment, in step (6), solution E is concentrated (e.g., by vacuum concentration, preferably by evaporation at about 50°C to 70°C, preferably about 54°C to 56°C) and then filtered to obtain an extract containing the active ingredient. Step (6) may be performed under a nitrogen atmosphere.

[0037] As used herein, the terms "cancer therapy," "anti-tumor therapy," "anti-cancer therapy," and "tumor therapy" can be used interchangeably and refer to a treatment method administered to a patient for cancer or tumor. Cancer therapy includes drug therapy and radiation therapy, of which drug therapy includes chemotherapy. These therapies can cause side effects in patients, including hematopoietic dysfunction (e.g., bone marrow suppression), gastrointestinal symptoms (e.g., nausea and vomiting), toxicity (e.g., cardiac toxicity and respiratory toxicity), and hair loss.

[0038] As used herein, the terms "anticancer drug therapy," "anticancer agent," and "antitumor agent" can be used interchangeably and refer to drug treatment administered to a patient's cancer or tumor. "Anticancer agents" can include cytotoxic agents and non-cytotoxic agents, and the cytotoxic agents can include agents that act directly on nucleic acids (e.g., DNA or RNA), antimetabolites (e.g., agents that inhibit DNA synthesis), and agents that act on structural proteins. In one embodiment, the anticancer agent can include an alkylating agent, and the alkylating agent can include nitrogen mustards. Cyclophosphamide belongs to the nitrogen mustard class of anticancer agents.

[0039] The "hematopoietic system" generally refers to the entire system that produces blood in the body, which is generally composed of two parts: hematopoietic organs and hematopoietic cells, and mainly includes the yolk sac, liver, spleen, kidneys, thymus, lymph nodes, and bone marrow. In the present invention, the hematopoietic system that is damaged by cancer treatment may be related to the bone marrow, liver, spleen, and lymph nodes, preferably the bone marrow.

[0040] As used herein, the terms "bone marrow suppression" and "bone marrow function suppression" can be used interchangeably and can refer to a decrease in the activity or number of blood cell precursors in the bone marrow, such as those caused by cancer treatment. Blood cells in the bloodstream have a short lifespan and require constant replenishment. To achieve this timely replenishment, stem cells, the precursors of blood cells, must divide rapidly. Drug treatments (e.g., cyclophosphamide), radiation therapy, and many other anti-tumor treatments all target rapidly dividing cells and therefore frequently suppress normal bone marrow cells. Among the myelosuppression caused by many cancer treatments, a relatively rapid and predominant decrease in white blood cells is usually the main cause. Therefore, the occurrence of myelosuppression can be determined by testing white blood cell counts after chemotherapy, and the effectiveness of a drug in restoring white blood cells is also an important indicator of the therapeutic efficacy of that drug in treating myelosuppression.

[0041] "Leukopenia": Usually, the number of white blood cells in the peripheral blood is continuously 4 x 10 9 Symptoms can appear when the white blood cell count is lower than 1 / L. While cancer treatment kills tumor cells, it also severely damages normal cells, especially those with high proliferation rates, such as bone marrow hematopoietic cells, resulting in a decrease in blood cells. Clinically, a decrease in white blood cells can lead to serious infections and other problems that often hinder the smooth progression of cancer therapy, thereby reducing the effectiveness of treatment and the patient's quality of life. Therefore, increasing peripheral white blood cells in cancer treatment has become a key factor in ensuring the completion and effectiveness of cancer treatment.

[0042] "Pancytopenia" refers to a medical condition of a decrease in the number of red blood cells, white blood cells, and platelets in the blood, including cytopenia caused by hematopoietic dysfunction or bone marrow suppression. Because the decrease in white blood cells is the main and most significant cause of cytopenia during cancer treatment, the effect of a drug on white blood cell counts can reasonably be interpreted as the effect on pancytopenia.

[0043] A side effect of cancer treatment can cause weight loss in patients, and therefore, the effect of a drug on weight gain in patients can also be an indicator of the effective alleviation or treatment of such side effects (e.g., hematopoietic system damage, bone marrow suppression, pancytopenia, and leukopenia). [Brief explanation of the drawings]

[0044] [Figure 1] Change curve of the effect of reconstitution on peripheral blood leukocytes in mice exposed to cyclophosphamide (250 mg / kg). WBC: White blood cells. [Figure 2] Effect of reconstitution on the amplitude of the increase in peripheral blood leukocyte concentration in mice exposed to cyclophosphamide (day 6). WBC: white blood cells. DETAILED DESCRIPTION OF THE INVENTION

[0045] Unless otherwise defined, all scientific terms used herein shall have the same meaning as commonly understood by one of ordinary skill in the art. Exemplary methods and materials are set forth below; equivalents may also be used. All publications and other references mentioned herein are incorporated by reference in their entirety.

[0046] The present invention will be described in more detail with reference to the following examples, which are not intended to limit the scope of the present invention in any way.

[0047] Example

[0048] Example 1 - Protective effect of rheumatoid arthritis on hematopoietic damage caused by cytotoxic drugs

[0049] 1. Purpose of the experiment We will create a hematopoietic system damage model using mice exposed to cyclophosphamide, observe the effects of repopulation on peripheral blood leukocytes in the model mice, and consider whether repopulation has the effect of promoting hematopoietic function in mice.

[0050] 2. Experimental Materials 2.1 Study drug This was a re-establishment project provided by Weishi Pharmaceutical (Rugao) Co., Ltd.

[0051] 2.2 Positive control Nilestriol was provided by Department 9 of the Institute of Radiation Medicine, Academy of Military Medical Sciences.

[0052] 2.3 Experimental animals The adult KM mice were bred by the Experimental Animal Center of the Academy of Military Medical Sciences. The mice weighed 18.0–20.0 g. The experimental animal permit number was SCXK-(Military) 2002-001. The experimental animals were maintained in the Animal Laboratory of the Academy of Military Medical Sciences, with a laboratory permit number of SYXK(Military)-2002-016. Ten mice were housed per cage and fed a specially formulated mouse diet with free access to water. The temperature in the animal laboratory was maintained at approximately 25°C, the relative humidity was maintained at 40–70%, and the lighting hours were 12 h daily.

[0053] 3. Experimental Method 3.1 Grouping Method Six groups were included in the experiment: a model control group, a positive control group (10 mg / kg nylestriol), a high-dose re-orientation group (14.4 U / kg), a medium-high-dose re-orientation group (7.2 U / kg), a medium-dose re-orientation group (3.6 U / kg), and a low-dose re-orientation group (1.8 U / kg). Mice were randomly grouped according to the mean normal blood white blood cell count before the two doses, with 10 mice per group.

[0054] 3.2 Model creation method Mice were administered a single dose of cyclophosphamide (250 mg / kg), the administration volume was 200 μl, and the administration route was all intraperitoneal injection.

[0055] 3.3 Administration method The positive control drug, nylestriol (10 mg / kg), was administered once 6 hours after cyclophosphamide administration in a volume of 200 μL. Administration to each reconstitution group began 6 hours after cyclophosphamide administration. Administration was continued every 48 hours for 2 weeks by intramuscular administration.

[0056] 3.4 Testing metrics for drug efficacy in mice Before creating the model, the mice were examined for normal blood status indicators, and after cyclophosphamide administration, white blood cell indicators were examined and measured every 2-3 days until the blood status was fully recovered. After the administration, the mice were sacrificed, and bone marrow sections were prepared and preserved in formalin to observe changes in bone marrow tissue.

[0057] 3.5 Data Processing The quantitative data were expressed as mean values ​​± standard deviation (x) and statistical analysis between groups was performed using the t-test program in Excel software. Change curves of various leukocyte indices were plotted using GraphPad Prism5.

[0058] 4. Experimental Results 4.1 Effects on mouse leukocytes One day after exposure to cyclophosphamide, white blood cell counts fell to their lowest value, after which recovery began. Compared with the model control group, the recovery rate was relatively fast in the reconstituted treatment group. Each group returned to near pre-treatment normal values ​​by day 12. See Table 1 and Figure 1 for details.

[0059] Table 1 shows the effect of reactivation on peripheral blood leukocytes in mice exposed to cyclophosphamide (250 mg / kg).

[0060] [Table 1]

[0061] vs: model control, "*" P<0.05; "**" P<0.01; "***" P<0.001

[0062] 5. Evaluation During the observed time period, the changes in the leukocyte counts in the animals' peripheral blood were relatively large. One day after cyclophosphamide administration, the leukocyte concentration in the model control group fell to its lowest value, then slowly recovered, approaching the pre-administration level 12 days after model creation.

[0063] The administration was performed 6 hours after the administration of cyclophosphamide. As shown in Table 1 and Figure 1, on the 6th day after model creation, the increase in leukocyte concentration in the test drug group was greater than that in the control group, suggesting that the test drug has a promoting effect on the recovery of the hematopoietic system. Figure 2 more directly demonstrates the promoting effect of repopulation on hematopoietic recovery.

[0064] 6. Conclusion In the hematopoietic recovery stage, the test drug was shown to effectively promote the recovery of the white blood cell index in mice. This further reasonably indicated that Li Zaishi could effectively treat hematopoietic system injury and myelosuppression induced by cancer treatment.

[0065] Example 2 - Effect of Li Zaishi on weight loss caused by cytotoxic drugs for relapse prevention

[0066] 1. Experimental purpose Using mice exposed to cyclophosphamide, a bone marrow injury model was established to observe the effect of the test drug on the body weight of the model mice.

[0067] 2. Experimental materials 2.1 Test drug It was Li Zaishi provided by Weishi Pharmaceutical (Rugao) Co., Ltd.

[0068] <00​​​​​​​​​​​​​​​​​​The remaining KM mice were used for model creation by intraperitoneal injection of cyclophosphamide (300 mg / kg). On the third day after the model was created, the blood status of the model KM mice was examined and measured, and the KM mice were randomly grouped according to their blood leukocyte counts, with 10 animals per group.

[0071] 3.2 Grouping Method The grouping and treatment methods are shown below. 1) Blank control group, 7 animals. 2) Model control group, 10 animals. 3) High-dose group suitable for reestablishment, intravenous administration, dose 14.4 U / kg, 10 animals. 4) Low-dose group for reestablishment, intravenous administration, dose 1.8U / kg, 10 animals.

[0072] 3.3 Administration method Administration began on the second day (fourth day after model preparation) when each test drug group was grouped, and was administered once every 48 hours for a total of four doses.

[0073] 3.4 Weight Inspection and Measurement The body weight of the KM mice was measured on day 3 before administration (-3d) and on days 1, 3, 5, 8, and 10 after administration (1d, 3d, 5d, 8d, and 10d).

[0074] 3.5 Statistical methods The quantitative data were expressed as X±S and statistical analysis was performed using Student's t-test.

[0075] 4. Results of efficacy experiments The body weight of all experimental animals in each model group was reduced, significantly lower than that of the blank control group (P<0.05). From the 5th day of treatment, body weight gradually recovered, and from the 8th day of treatment, body weight clearly recovered, with significant differences compared with the model control group (P<0.05). See Tables 2 and 3.

[0076] Table 2 shows the weight changes of the KM model mice and mice in each treatment group.

[0077] [Table 2]

[0078] Note: Compared with the blank control group, "*" P<0.05; "**" P<0.01; "***" P<0.001; compared with the model control group, "?" P<0.05; "??" P<0.01; "???" P<0.001.

[0079] Table 3. Relative weight gain of KM model mice and mice in each treatment group

[0080] [Table 3]

[0081] Note: Compared with the blank control group, "*" P<0.05; "**" P<0.01; "***" P<0.001; compared with the model control group, "?" P<0.05; "??" P<0.01; "???" P<0.001.

[0082] 5. Experimental Evaluation In a mouse model of bone marrow hematopoietic damage caused by cyclophosphamide, all mice showed weight loss. The test drug had a promoting effect on weight recovery in mice, and there was a statistically significant difference compared with the model control group.

[0083] 6. Conclusion The experimental drug re-application was able to promote weight recovery in a mouse model of bone marrow injury caused by cyclophosphamide, indirectly suggesting that this could effectively alleviate hematopoietic damage and bone marrow suppression caused by anti-cancer treatment.

Claims

1. 1. Use of an extract from rabbit skin inflamed by vaccinia virus in the manufacture of a medicament for the prevention or treatment of leukopenia induced by cancer therapy in a patient, comprising: The cancer treatment is a treatment involving administration of cyclophosphamide.

2. 1. Use of an extract derived from rabbit skin inflamed by vaccinia virus in the manufacture of a medicament for restoring or increasing white blood cell count in a patient undergoing cancer treatment, comprising: The cancer treatment is a treatment involving administration of cyclophosphamide.

3. The use according to claim 1 or 2, wherein the extract derived from rabbit skin inflamed by vaccinia virus is formulated into an oral or injectable preparation.

4. The use according to claim 3, wherein the injection is an intramuscular injection or an intravenous injection.

5. The use according to any one of claims 1 to 4, wherein the medicament comprises 0.6 U to 300 U of an extract derived from rabbit skin inflamed by the vaccinia virus.

6. The use according to claim 5, wherein the medicament comprises an extract derived from rabbit skin inflamed by 9 U to 70 U of the vaccinia virus.

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