WO2021164706A1 - Composition pharmaceutique contenant un colorant bleu de méthylène et son utilisation - Google Patents

Composition pharmaceutique contenant un colorant bleu de méthylène et son utilisation Download PDF

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Publication number
WO2021164706A1
WO2021164706A1 PCT/CN2021/076749 CN2021076749W WO2021164706A1 WO 2021164706 A1 WO2021164706 A1 WO 2021164706A1 CN 2021076749 W CN2021076749 W CN 2021076749W WO 2021164706 A1 WO2021164706 A1 WO 2021164706A1
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WIPO (PCT)
Prior art keywords
methylene blue
tumor
local
blue dye
composition
Prior art date
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PCT/CN2021/076749
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English (en)
Chinese (zh)
Inventor
邹方霖
邹礼常
王建霞
王艺羲
Original Assignee
成都夸常奥普医疗科技有限公司
夸常股份有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Publication date
Priority claimed from PCT/CN2020/075765 external-priority patent/WO2021163897A1/fr
Priority claimed from CN202011059699.0A external-priority patent/CN114344333A/zh
Priority claimed from CN202011064448.1A external-priority patent/CN114306392A/zh
Priority claimed from CN202011059746.1A external-priority patent/CN114306612A/zh
Application filed by 成都夸常奥普医疗科技有限公司, 夸常股份有限公司 filed Critical 成都夸常奥普医疗科技有限公司
Publication of WO2021164706A1 publication Critical patent/WO2021164706A1/fr
Priority to EP21874572.7A priority Critical patent/EP4223313A1/fr
Priority to US18/247,359 priority patent/US20230414617A1/en
Priority to JP2023519733A priority patent/JP2023543858A/ja
Priority to PCT/CN2021/122041 priority patent/WO2022068918A1/fr
Priority to PCT/CN2022/076823 priority patent/WO2022174812A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/255—Esters, e.g. nitroglycerine, selenocyanates of sulfoxy acids or sulfur analogues thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
    • A61K31/5415—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame ortho- or peri-condensed with carbocyclic ring systems, e.g. phenothiazine, chlorpromazine, piroxicam
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • 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 disclosure of this application relates to a composition containing the methylene blue dye and its local synergist or synergistic drug and its use in the preparation of a drug combination or preparation for cancer.
  • the present invention relates to methylene blue dyes and their local synergistic drugs or synergistic compositions that preferably minimize rather than maximize local effects as active ingredients that can provide local synergistic effects when prepared to provide said local synergistic effects.
  • a topical pharmaceutical composition for the treatment of cancer such as malignant solid tumors, comprising the methylene blue dye and a local synergistic drug or synergist thereof that can provide a local synergistic effect for the treatment of malignant solid tumors Drugs, and methods for treating cancer such as malignant solid tumors that include administering the pharmaceutical composition or drug combination or formulation.
  • anti-solid tumor drugs One of the main problems facing the development of anti-solid tumor drugs is specificity. Since conventional anti-tumor drugs cannot sufficiently distinguish between target cells and normal cells, and the difference between the effective dose and the safety limit is not large enough, they will produce systemic effects (tumor cell inhibitory effects inside and outside the tumor) at the same time. Greater risk of systemic toxicity. In addition, the drug molecule needs to penetrate effectively in the tumor tissue to have an effect on the tumor cells in between. For some tumors with poor blood supply (such as pancreatic cancer), the chance of benefiting the patient is even smaller.
  • Intratumoral administration has the advantage of physically targeting drugs.
  • intratumoral administration of conventional anti-tumor drugs increased the concentration of the target area, it did not show a significant improvement in efficacy.
  • conventional anti-tumor drugs are almost still administered systemically in clinical practice.
  • Conventional chemical ablation agents high-purity ethanol, high-concentration acids and bases
  • the target tissue cannot be sufficiently distinguished from other tissues, the actual intervention volume (for example, the amount of acid and alkali does not exceed 0.2 ml/kg) and the intervention site are very limited. Therefore, chemical ablation agents have gradually faded out of malignant solid tumors in the past ten years. In fact, there are almost no local special drugs with high local safety and high local curative effect in clinical practice.
  • the purpose of the present invention is to provide a physical target tumor body, but has a higher curative effect, higher compliance, or higher compliance than existing drugs (especially pharmaceutical compositions containing methylene blue dyes in the prior art) / And a more specific topical drug, and a method for treating malignant solid tumors including administering the pharmaceutical composition.
  • a pharmaceutical composition comprising a methylene blue dye as a topical active ingredient and a local synergist or a synergistic drug of the methylene blue dye, wherein the synergist or synergistic drug comprises or One or more selected from nutrients, immunomodulators, and conventional anti-tumor drugs.
  • a combination comprising methylene blue dye, a topical synergist or a synergistic drug, and optionally a nutrient or/and an immunomodulator, wherein the synergistic or synergistic drug includes or One or more selected from conventional anti-tumor drugs, wherein in the composition, the concentration (w/v) of the methylene blue dye when administered is ⁇ 2%, preferably 0.35-2% , 0.5-2%, 0.5-1.5% or 0.5-1%.
  • the concentration (w/v) of the methylene blue dye when applied is ⁇ 2%, for example, 0.35%, 0.5%, 0.7%, 0.9%, 1%, 1.25%, 1.5% , 1.8%, 2%, or any range therebetween, preferably 0.35-2%, 0.5-2%, 0.5-1.5%, 0.7-1.5%, 0.7-1.25% or 0.5-1%.
  • composition comprising a methylene blue dye and a topical synergist or a synergistic drug, wherein the synergist or synergistic drug includes or is selected from one or more nutrients, and Optional conventional anti-tumor drugs or/and immunomodulators.
  • composition comprising a methylene blue dye and a local synergist or a synergistic drug, wherein the synergist or synergistic drug includes or is selected from one or more immunomodulators.
  • the synergist or synergistic drug includes or is selected from one or more immunomodulators.
  • a composition comprising methylene blue dye and a topical synergist or a synergistic drug, wherein the synergist or synergistic drug includes or is selected from nutrients, wherein in the composition, the concentration of the methylene blue dye when applied is a concentration (w/v) ⁇ 2%, preferably 0.35-2%, 0.5-2%, 0.5-1.5% or 0.5-1%.
  • the methylene blue dye may include methylene blue and its living dye analogs, preferably, include a compound selected from the group consisting of methylene blue, patent blue, isosulfan blue, Neomethylene blue is more preferably selected from methylene blue and its derivatives.
  • the methylene blue dye may be selected from methylene blue and its living dye analogs, preferably, it may be selected from methylene blue, patent blue, isosulfur blue, neomethylene blue, more preferably Ground is selected from methylene blue and its derivatives.
  • the nutrient includes one or more of the following: amino acid nutrient, carbohydrate nutrient, lipid nutrient.
  • the nutrient is selected from one or more of amino acid nutrients, carbohydrate nutrients, and lipid nutrients.
  • the concentration of the above-mentioned nutrients at the time of administration is 2.5-50%, preferably 4-40%.
  • the nutrient includes DHA.
  • the amino acid nutrient includes or is selected from one or more of the following amino acid compounds with nutritional and health effects: amino acids, amino acid salts, oligopeptides and polypeptides; preferably amino acids or their salts selected from the following Or oligopeptides and polypeptides containing or consisting of the following amino acids: alanine, valine, leucine, isoleucine, phenylalanine, proline, tryptophan, tyrosine, serine, half Cystine, methionine, asparagine, glutamine, threonine, lysine, arginine, histidine, aspartic acid, glutamic acid, ⁇ -alanine, taurine, ⁇ Aminobutyric acid (GABA), tea polyphenols (theanine), pumpkin seed amino acids (3-amino-3-carboxypyranoic acid), glutamine, citrulline, ornithine; more preferably selected from the following The amino acid or its salt or
  • the concentration of the amino acid compound when administered is 2.5-50%, preferably 4-40%.
  • the amino acid nutrient is selected from amino acids or amino acid salts with nutritional and health effects, and in the composition, the concentration (w/v) of the amino acid or amino acid salt at the time of administration is ⁇ 2%, ⁇ 2.5, ⁇ 5%, ⁇ 7.5%, 10-25% or 18-25%, preferably 15%-25% or 20%-25%.
  • the amino acid nutrients are selected from oligopeptides and polypeptides with nutritional and health effects, and in the composition, the concentration (w/v) of the oligopeptides and polypeptides at the time of administration is greater than ⁇ 5% , Preferably 7.5-25%, more preferably 10%-25%.
  • the amino acid nutrient is a combination of the amino acid and/or amino acid salt and the oligopeptide and/or polypeptide, and in the pharmaceutical composition, the concentration of the combination at the time of administration (w/v) It is greater than ⁇ 5%, preferably 7.5%-25%, more preferably 10-25%.
  • the oligopeptide includes or is selected from one or more of the following: glycyl-L-tyrosine, glycylalanine, glycylglycine, lysine-glycine dipeptide, Gamma dipeptide, carnosine ( ⁇ -alanine histidine copolymer), glutathione, collagen oligopeptides, casein hydrolyzed peptides, soybean oligopeptides, oligoarginine, oligoglycine, oligolysine
  • the polypeptide is one or more selected from polyaspartic acid, polyglutamic acid, and polylysine.
  • the carbohydrate nutrient includes or is a carbohydrate compound containing one or more of the following sugar units: glucose, ribose, xylose, fructose, galactose, fucose, preferably one selected from the following Species or more: glucose, fructose, chitooligosaccharides, glucosamine, lactulose, sorbitol, ribose, sorbose, mannose, galactose, sucrose, lactose, trehalose, xylo-oligosaccharides, fructo-oligosaccharides, mannose oligosaccharides Sugar, gluconic acid, sodium gluconate, xylitol, mannitol, maltitol, lactitol, more preferably one or more selected from the following: glucose, sodium gluconate, chitooligosaccharide, glucosamine, lactulose , Ri
  • the concentration (w/v) of the carbohydrate nutrient at the time of administration is greater than 5%, preferably ⁇ 10%, 10-40%, 15-50% or 25-50% .
  • the lipid nutrient includes or is selected from one or more of the following: fatty acid, fat milk and lipid, preferably includes or is selected from one or more of the following: vegetable oil, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), long-chain fat emulsion, medium-chain fat emulsion, phospholipid, and in the composition, the concentration of the lipid nutrient when administered is ⁇ 4%, preferably It is 4%-25%.
  • EPA eicosapentaenoic acid
  • DHA docosahexaenoic acid
  • the concentration of the lipid nutrient when administered is ⁇ 4%, preferably It is 4%-25%.
  • the composition of the present invention also includes conventional anti-tumor drugs.
  • the anti-tumor drugs include or are selected from one or more of the following groups: drugs that disrupt the structure and function of DNA, and intervene in DNA to interfere with transcription of RNA. Drugs, drugs that interfere with DNA synthesis, drugs that affect protein synthesis.
  • conventional anti-tumor drugs include or are selected from one or more of the following: uracil derivatives, cyclophosphamides, gemcitabine, epirubicin, anti-tumor antibiotics, tenipo Glycosides, metal platinum complexes, taxanes, preferably one or more selected from the following drugs and their analogous derivatives: 5-fluorouracil, cyclophosphamide, gemcitabine, epirubicin, antitumor antibiotics, and Niposide, metal platinum complex, paclitaxel.
  • composition of the present invention further includes an immunomodulator, which includes or is selected from one or more of antibodies, nucleic acids, and probiotic components.
  • the concentration (w/v) of the methylene blue dye when applied is ⁇ 2%, preferably 0.35-2%, 0.5-2%, 0.5-1.5% or 0.5-1%, so
  • the concentration (w/v) of the nutrient at the time of administration is ⁇ 2%, preferably 3-40%, and the concentration of the conventional anti-tumor drug at the time of administration is greater than 20% or 30% of its saturated concentration, preferably its saturated concentration
  • the concentration is 30%-100%, wherein the saturated concentration refers to the saturated concentration of the conventional anti-tumor drug in the liquid carrier.
  • the present invention includes immunomodulators, such as probiotic components, immunoglobulins, immunomodulatory peptides.
  • immunomodulators such as probiotic components, immunoglobulins, immunomodulatory peptides.
  • the strains of probiotics used are all strains used in Saccharomyces cerevisiae and probiotic preparations approved by the food and drug authorities or contained in the official pharmacopoeia.
  • strains contained in probiotic preparations approved by the Chinese pharmaceutical authority Clostridium butyricum in Bacillus coagulans live tablets, Bacillus licheniformis strains in live Bacillus licheniformis granules (capsules), Oral Bacillus cereus Bacillus cereus strains in live bacteria preparations, Clostridium butyricum strains in live Clostridium butyricum capsules, Saccharomyces boulardii strains in Saccharomyces boulardii powder (capsules), Lactobacillus acidophilus capsules Lactobacillus butyricum strain, Clostridium butyricum in oral live Clostridium butyricum powder, Enterococcus faecium strain, subtilis strain, and Enterococcus butyricum triple viable tablet in Enterococcus subtilis double live bacteria multidimensional granules Lactobacillus strains, Clostridium butyricum, saccharification strains, Lacto
  • composition of the present invention optionally further comprises one or more of the following: analgesic, sustained-release carrier, and C1-10 aliphatic optionally substituted with 1-3 hydroxyl groups as an acidulant carboxylic acid.
  • the analgesic agent is one or more including or selected from the following: benzyl alcohol, procaine hydrochloride, chlorobutanol, and lidoca hydrochloride, and in the composition, the The concentration of the analgesic at the time of administration is 0.1-4% by weight.
  • the acidulant is one or more selected from the group consisting of acetic acid, propionic acid, butyric acid, malonic acid, succinic acid, glycolic acid, lactic acid, citric acid, malic acid, tartaric acid, More preferred is acetic acid.
  • the composition of the present invention further includes a pharmaceutically acceptable carrier, such as an aqueous carrier or an alcohol carrier, preferably water and/or ethanol.
  • a pharmaceutically acceptable carrier such as an aqueous carrier or an alcohol carrier, preferably water and/or ethanol.
  • the composition of the present invention may be prepared as an injection dosage form, which includes a liquid injection and a powder injection for injection.
  • the injection powder for injection includes a sterile dry powder and a solvent, and one or all of the amino acid nutrients and the ineffective absorption compound are contained in the sterile dry powder, and the liquid carrier is contained in the sterile dry powder.
  • the solvent, and the concentration of the amino acid nutrients and the ineffective absorption compound are their concentrations in the sterile dry powder and the solvent mixture, respectively.
  • the ratio of the amount of the methylene blue dye to the nutrient, immunomodulator, and anti-tumor drug is selected from one or more of the following groups: partially synergistic ratio (W nutrients / W methylene blue dye) is 1/3 ⁇ W nutrient / W methylene blue dye ⁇ 40 / 0.15, preferably 1/3 ⁇ W nutrient / W type methylene blue dye ⁇ 40 / 0.30, or 1 / 1.8 ⁇ W nutrient / W methylene blue dye ⁇ 40 / 0.30; synergistic ratio of the partial immunomodulator / methylene blue dye composition (W immunomodulators / W methylene blue dye) was 0.2 / 3 ⁇ W immunomodulators / W methylene blue dye ⁇ 20 / 0.15, preferably from 0.5 / 3 ⁇ W immunomodulators / W type methylene blue dye ⁇ 20 / 0.50, or 0.5 / 1 ⁇ W immunomodulators / W methylene blue dye ⁇ 20
  • a medical device comprising a composition according to the above is provided.
  • the application of the above-mentioned composition and an anti-tumor drug or immunomodulator in the preparation of a drug combination or preparation for the treatment of cancer is provided.
  • the cancer includes solid tumors, such as sarcoma, head and neck cancer, breast cancer, pancreatic cancer, thyroid cancer, nasopharyngeal cancer, prostate cancer, liver cancer, lung cancer, bowel cancer, oral cancer, esophageal cancer, gastric cancer, Laryngeal cancer, testicular cancer, vaginal cancer, uterine cancer, ovarian cancer, etc.
  • solid tumors such as sarcoma, head and neck cancer, breast cancer, pancreatic cancer, thyroid cancer, nasopharyngeal cancer, prostate cancer, liver cancer, lung cancer, bowel cancer, oral cancer, esophageal cancer, gastric cancer, Laryngeal cancer, testicular cancer, vaginal cancer, uterine cancer, ovarian cancer, etc.
  • composition of the present invention may be used for local administration, for example, local intratumoral administration.
  • a method for treating cancer which includes administering the above-mentioned composition or a pharmaceutical combination or preparation thereof with an anti-tumor drug or an immunomodulator to a patient suffering from cancer.
  • a method for treating cancer which includes intratumorally administering the above-mentioned composition or a pharmaceutical combination or preparation thereof with an antitumor drug or an immunomodulator to a patient suffering from cancer.
  • the cancer includes solid tumors, such as sarcoma, head and neck cancer, breast cancer, pancreatic cancer, thyroid cancer, nasopharyngeal cancer, prostate cancer, liver cancer, lung cancer, bowel cancer, oral cancer, esophageal cancer, gastric cancer, Laryngeal cancer, testicular cancer, vaginal cancer, uterine cancer, ovarian cancer, etc.
  • the solid tumor is preferably selected from malignant solid tumors containing tumor bodies with a mesenchymal ratio ⁇ 20% or/and an average size ⁇ 2.0 cm.
  • a combination of a local synergist or a synergistic drug of a methylene blue dye and the methylene blue dye which is preferably to minimize rather than maximize local effects, is provided as a local synergistic effect.
  • the methylene blue dye is used as the topical active ingredient
  • the local synergist or synergistic drug is selected from the following One or more of the group: nutrients, immunomodulators, conventional anti-tumor drugs.
  • a topical pharmaceutical composition for the treatment of malignant solid tumors that can provide the local synergistic effect, which comprises a methylene blue dye that preferably minimizes rather than maximizes the local effect, and
  • the local synergist or synergistic drug of the methylene blue dye wherein the methylene blue dye is used as a local active ingredient, and the synergist or synergistic drug is selected from one or more of the following groups: nutrients, immunomodulators Agents, conventional anti-tumor drugs.
  • the ratio of the amount of the methylene blue dye to the nutrient, immunomodulator, and anti-tumor drug is selected from one or more of the following groups: partially synergistic ratio (W nutritional factors / W methylene blue dye) is 1/3 ⁇ W nutrient / W methylene blue dye ⁇ 40 / 0.15, preferably 1/3 ⁇ W nutrient / W methylene blue dye ⁇ 40/0.30, or 1/1.8 ⁇ W nutrient /W methylene blue dye ⁇ 40/0.30; the local synergistic amount ratio of the immunomodulator/methylene blue dye composition (W immunomodulator /W sub A blue dye) was 0.2 / 3 ⁇ W immunomodulators / W methylene blue dye ⁇ 20 / 0.15, preferably from 0.5 / 3 ⁇ W immunomodulators / W methylene blue dye ⁇ 20 / 0.50, or 0.5 /1 ⁇ W immunomodulator /W methylene blue dye ⁇ 20/0.50; the local synergistic ratio
  • the above-mentioned topical pharmaceutical composition further comprises a pharmaceutically acceptable liquid carrier when administered, and in the administration composition of the topical pharmaceutical composition, the concentration of the methylene blue dye (w/v ) ⁇ 2%, preferably 0.35-2%, 0.5-2%, 0.5-1.5% or 0.5-1%, the concentration of the nutrient is greater than 2%, preferably 3%-40%, the immunomodulator
  • concentration (w/v) is greater than 0.25%, preferably 0.5%-30%
  • the concentration (w/v) of the conventional anti-tumor drug is greater than 20% of its saturated concentration, preferably 30% of its saturated concentration -100%, wherein the saturated concentration refers to the saturated concentration of the conventional anti-tumor drug in the liquid carrier.
  • a topical pharmaceutical composition for the treatment of malignant solid tumors which comprises (preferably to minimize rather than maximize local effects) methylene blue Dyes, local synergists or synergistic drugs of the methylene blue dyes, and pharmaceutically acceptable liquid carriers, wherein the synergists or synergistic drugs include or are selected from one or more of the following: nutrients , Immunomodulators, conventional anti-tumor drugs, and in the administration composition of the topical pharmaceutical composition, the concentration (w/v) of the methylene blue dye is ⁇ 2%, preferably 0.35-2%, 0.5- 2%, 0.5-1.5% or 0.5-1%, the concentration of the nutrient is greater than 2%, preferably 3%-40%, and the concentration (w/v) of the immunomodulator is greater than 0.25%, preferably 0.5%-30%, and the concentration (w/v) of the conventional antitumor drug is greater than 20% of its saturated concentration, preferably 30%-100% of
  • the methylene blue dye is selected from methylene blue and its living dye analogs, preferably selected from the following compounds and their derivatives: methylene blue, patent blue, isosulfur blue, neomethylene blue , More preferably selected from methylene blue and its derivatives, wherein the concentration (w/v) of the methylene blue dye is ⁇ 1.1%, preferably 0.35-1.09%, 0.5-1.09% or 0.5- 1.05%.
  • the above-mentioned nutrients are one or more selected from the following group: amino acid nutrients, carbohydrate nutrients, lipid nutrients, and wherein the concentration of the nutrients is greater than 2.5% to less than 50%, preferably 4-40%.
  • the above-mentioned immunomodulator is one or more selected from the group consisting of antibodies, nucleic acids, and probiotic components, and wherein the concentration (w/v) of the immunomodulator is greater than 0.25%, Preferably it is 0.5%-30%.
  • the above-mentioned conventional anti-tumor drugs are selected from one or more of the following groups: drugs that disrupt the structure and function of DNA, drugs that intercalate in DNA and interfere with transcription of RNA, drugs that interfere with DNA synthesis, and drugs that affect protein synthesis .
  • the above-mentioned pharmaceutical composition is in the form of an injection, and the injection includes a liquid injection and a powder for injection.
  • a topical pharmaceutical composition for treating malignant solid tumors which comprises a dry powder obtained by freeze-drying or semi-freeze-drying part or all of the above-mentioned pharmaceutical composition.
  • the malignant solid tumors include breast cancer, pancreatic cancer, thyroid cancer, nasopharyngeal cancer, prostate cancer, liver cancer, lung cancer, bowel cancer, oral cancer, esophageal cancer, stomach cancer, laryngeal cancer, testicular cancer, vagina Cancer, uterine cancer, ovarian cancer.
  • the malignant solid tumor is preferably selected from malignant solid tumors containing tumor bodies with a mesenchymal ratio ⁇ 20% or/and an average size ⁇ 2.0 cm.
  • a topical pharmaceutical composition for the treatment of malignant solid tumors which comprises a methylene blue dye, a local synergistic drug or synergist of the methylene blue dye, and pharmacology
  • An acceptable liquid carrier wherein the synergistic drug or synergist is selected from nutrients or/and conventional anti-tumor drugs, and in the topical pharmaceutical composition, the concentration of the methylene blue dye (w/v) ⁇ 2 %, preferably 0.35-2%, 0.5-2%, 0.5-1.5% or 0.5-1%, the concentration of the nutrient (w/v) ⁇ 2%, preferably 3-40%, and the conventional antibiotic
  • the concentration of the tumor drug is greater than 20% or 30% of its saturated concentration, preferably 30%-100% of its saturated concentration, wherein the saturated concentration refers to the saturated concentration of the conventional anti-tumor drug in the liquid carrier.
  • a topical active ingredient as a topical active ingredient and a local synergistic drug or synergist of the methylene blue dye to prepare a topical pharmaceutical composition for the treatment of malignant solid tumors
  • the synergistic drugs or synergists are selected from nutrients or/and conventional anti-tumor drugs.
  • the topical pharmaceutical composition comprises the methylene blue dye, a local synergistic drug or synergist of the methylene blue dye, and a pharmaceutically acceptable liquid carrier, wherein
  • the concentration (w/v) of the methylene blue dye is ⁇ 2%, preferably 0.35-2%, 0.5-2%, 0.5-1.5% or 0.5-1%, and the nutrient
  • the concentration (w/v) of ⁇ 2%, preferably 3%-40%, and the concentration of the conventional antitumor drug is greater than 20% or 30% of its saturated concentration, preferably 30%-100% of its saturated concentration
  • the saturated concentration refers to the saturated concentration of the conventional anti-tumor drug in the liquid carrier.
  • it provides a method for treating malignant solid tumors, which comprises intratumorally administering the pharmaceutical composition disclosed in the present application to an individual in need thereof.
  • composition containing the methylene blue dye disclosed in the present application also referred to as the composition of the invention in the present invention
  • the composition containing the methylene blue dye in the prior art also in the present invention
  • shared pharmacology is more optimized: the former preferentially utilizes the effective local synergy of the composition, while the latter preferentially utilizes the local effect of methylene blue dyes and additives to the local The additive synergistic effect of the effects; 2)
  • the effective technical solution to achieve shared pharmacology is more optimized: the former only needs to be the methylene blue dye that minimizes rather than maximizes the local effect, while the latter needs to be optimized for the maximum
  • the methylene blue dyes that do not minimize local effects are embodied in two opposite directions of preferred dosing concentration.
  • the side effects of the local synergistic drugs required by the former are not stronger (in terms of being selected from conventional antitumor drugs), or even weaker (in terms of being selected from nutrients or/and immunomodulators) than any of the latter.
  • the former can be used preferentially in one or more of the following indications that the latter cannot: a larger stromal ratio tumor that can be administered locally, and a tumor containing the tumor Treatment of tumors; treatment of larger tumors that can be administered locally and tumors containing the tumors; treatment of nodules related to malignant tumors that can be administered locally.
  • the embodiment according to the present invention has the following advantages: Compared with the existing cytotoxic drugs and related treatment methods, it shows almost non-toxic systemic safety and significantly higher localization.
  • the method and composition of the present invention are also not troubled by the drug resistance problems encountered by existing cytotoxic drugs and existing molecular targeted drugs.
  • the method and composition are convenient in application and low in cost, and are particularly helpful for the general population who cannot afford high expenses to enjoy safe and effective treatment.
  • the inventor of the present invention unexpectedly discovered in a tumor-bearing animal experiment that although the addition of DHA generally cannot, it can form a highly synergistic effect with methylene blue under certain specific conditions, which also makes the use of methylene blue
  • the dose can be reduced exponentially to reduce the risk of side effects.
  • substances selected from other nutrients or/and conventional anti-tumor drugs can also form an unexpected synergistic effect with lower concentrations (for example, ⁇ 1%) of methylene blue under these specific conditions.
  • These specific conditions are not the conditions of methylene blue dyes, nutrients or/and conventional anti-tumor drugs in the existing anti-malignant solid tumor technology (for example, intratumoral administration concentration), but are as defined below.
  • the term “pharmaceutical composition” refers to a substance that clarifies the pharmacology and provides the pharmacological methods, pharmacological composition, pharmacological environment and other pharmacological characteristics necessary for realizing the pharmacology in the patient's body.
  • the term “pharmacological method” refers to the method of administration of the pharmaceutical composition necessary for the realization of a specific pharmacology.
  • the pharmacological method necessary for the improvement of the intestinal barrier of probiotics is oral administration
  • the pharmacological method necessary for ethanol chemical ablation is administration in the lesion area.
  • pharmaceutical composition refers to the composition of the pharmaceutical composition necessary for the realization of a specific pharmacology.
  • pharmacological environment refers to the minimization of exogenous interference necessary for the pharmaceutical composition to achieve a specific pharmacology in the target area.
  • additives such as osmotic pressure regulators
  • Negative effects A pharmaceutical composition without a specific pharmacological environment is bound to contain inactive ingredients that enhance administration compliance (for example, flavoring agents for oral drugs) and administration safety (for example, osmotic pressure regulators contained in injections), while local effects Pharmacology must require a limited pharmacological environment to reduce the possible pharmacological interference of inactive ingredients.
  • the pharmacology of pharmaceutical compositions without specific pharmacological methods means that these methods can be achieved (e.g. probiotic injections for immune enhancement, cytotoxic drugs for cell growth inhibition)
  • the drunken reaction of ethanol means that it can only be achieved in this method (for example, the improvement of the intestinal barrier of oral probiotics, the chemical ablation of topical ethanol).
  • the pharmacology of a pharmaceutical composition without a specific pharmacological composition limitation means that the pharmacology of a pharmaceutical composition is in a given active ingredient, active ingredient amount ratio, and It can be achieved after the dosage of the composition (for example, the immune enhancement of probiotic injections, the cell growth inhibition of cytotoxic drugs, and the drunken reaction of ethanol).
  • pharmacology of ethanol under the limits of its pharmacological method (administration within the lesion), pharmacological concentration (for example, high concentration ⁇ 70%), and pharmacological volume (for example, dosing volume/target volume ratio ⁇ 0.25)
  • pharmacological concentration for example, high concentration ⁇ 70%
  • pharmacological volume for example, dosing volume/target volume ratio ⁇ 0.25
  • topical drug is distinguished from conventional drugs (compositions).
  • compositions refers to conventional administration (or systemic administration, such as oral, intravenous, intraperitoneal injection, pleural injection). Etc.) so that its active ingredients are transported to the tumor through the blood and diffuse and penetrate through the blood vessels (compositions).
  • Therapeutic drugs whose activity produces medicinal effects.
  • local activity refers to a pharmacological activity that is preferentially displayed in local tissues rather than tumor cells, and includes, for example, the local action of a single drug and the local shared action of a shared drug.
  • the term "locally active ingredient” is distinguished from chemical ablation agents, which refers to chemical substances (such as 50% acetic acid, absolute ethanol, 5% methylene blue) under effective ablation conditions of the tumor (usually exceeding the ablation concentration threshold) , While the former refers to Ya-Africa through its conventional effects (not any administration other than intratumoral administration), nor its chemical ablation effect (used under conditions lower than its chemical ablation concentration), but mainly by providing local Synergistic active ingredients (e.g. 0.5 to 1.5% methylene blue).
  • local synergistic drug (or local synergist) is used to refer to the local active ingredient (for example, local administration and administration concentration ⁇ 2%, preferably 0.5%-1.5%) Methylene blue dyes) share substances that produce local synergistic effects, which are selected from one or more of the following groups: nutrients, immunomodulators, and conventional anti-tumor drugs.
  • local synergistic effect refers to a synergistic effect that mainly exhibits local activity.
  • the term “synergistic effect” means that the sharing of active ingredients shows a more favorable pharmaceutical effect for treatment than they are used alone, and it includes, for example, synergistic efficacy and synergistic safety.
  • the term “synergistic effect” refers to the combination of active ingredients exhibiting a higher desired medicinal effect than any component used alone, and/or the combination of any component exhibiting no single use.
  • the drug effect is required (for example, the tissue necrosis effect produced by the sharing of two cytotoxic drugs, or the drug effect of the actual/expected ratio q>1.15 agreed in the embodiment of the present invention).
  • the term “synergistic safety” means that when effective pharmacological effects are obtained, the active components collectively show a higher required safety than any component used alone.
  • a topical pharmaceutical composition for the treatment of malignant solid tumors which comprises a methylene blue dye, a local synergistic drug or synergist of the methylene blue dye, and a drug A scientifically acceptable liquid carrier, wherein the synergistic drug or synergist is selected from nutrients or/and conventional anti-tumor drugs, and in the topical pharmaceutical composition, the concentration of the methylene blue dye (w/v) ⁇ 2%, preferably 0.35-2%, 0.5-2%, 0.5-1.5% or 0.5-1%, the concentration of the nutrient (w/v) ⁇ 2%, preferably 3%-40%, and the concentration of the conventional anti-tumor drug is greater than 20% or 30% of its saturated concentration, preferably 30%-100% of its saturated concentration, wherein the saturated concentration refers to the saturation of the conventional anti-tumor drug in the liquid carrier concentration.
  • a topical active ingredient as a topical active ingredient and a local synergistic drug or synergist of the methylene blue dye to prepare a topical pharmaceutical composition for the treatment of malignant solid tumors
  • the synergistic drugs or synergists are selected from nutrients or/and conventional anti-tumor drugs.
  • the topical pharmaceutical composition comprises the methylene blue dye, a local synergistic drug or synergist of the methylene blue dye, and a pharmaceutically acceptable liquid carrier, wherein
  • the concentration (w/v) of the methylene blue dye is ⁇ 2%, preferably 0.35-2%, 0.5-2%, 0.5-1.5% or 0.5-1%, and the nutrient
  • the concentration (w/v) of ⁇ 2%, preferably 3%-40%, and the concentration of the conventional antitumor drug is greater than 20% or 30% of its saturated concentration, preferably 30%-100% of its saturated concentration
  • the saturated concentration refers to the saturated concentration of the conventional anti-tumor drug in the liquid carrier.
  • it provides a method for treating malignant solid tumors, which comprises intratumorally administering a therapeutically effective amount of the methylene blue dye disclosed in the present application to an individual in need thereof.
  • intramoral administration refers to the injection of drugs (such as injections) into the tumor through devices, such as transcatheter arterial infusion, transcatheter intratumor infusion, intratumoral injection, and the like.
  • therapeutically effective amount refers to the amount of a drug used to treat a disease (such as a tumor) and obtain an effective effect (such as reducing or/and alleviating the symptoms of the disease).
  • concentration refers to the weight/volume percentage concentration% (w/v) of the specified component in the topical pharmaceutical composition.
  • intramoral administration concentration refers to the concentration of the specified component when the drug is administered intratumorally, which may be the concentration of the specified component where the drug contacts the target area (for example, the injection needle hole or the outlet of the perfusion tube).
  • methylene blue dyes at the above concentrations have many applications, such as antidote, analgesic, vital dyes, and so on.
  • a new application of methylene blue dyes in the prior art involves topical administration.
  • the methylene blue dyes in this application must maximize rather than minimize local effects pharmacologically, and therefore must be a maximum preferred concentration orientation in terms of composition.
  • synergists can be optionally added, which involve systemic administration of effective anti-tumor substances, Systemic administration of low-efficiency but local administration of high-efficiency low-efficiency anti-tumor substances and metal compounds.
  • a high concentration (for example, 5%) methylene blue dye can be used as a chemical ablation agent in this technical solution. It is generally believed that the greater the concentration of the chemical ablation agent, the more effective it is.
  • the inventors of the present invention unexpectedly discovered that the methylene blue dyes provide only conditions that preferably minimize rather than maximize local effects under non-chemical ablative conditions (for example, 1% methylene blue). Blue administration concentration), it can be shared with nutrients, immunomodulators, and/or conventional anti-tumor drugs that are quite different from pharmacologically.
  • the expected local synergy so that a methylene blue dye that is significantly lower than its chemical ablation concentration threshold can be used to obtain a significantly improved therapeutic effect.
  • These unexpected synergistic effects may be the key point of pharmacology, which significantly improves the specificity of methylene blue dyes, nutrients, immunomodulators, and/or conventional anti-tumor drug compositions against intratumoral tissue destruction.
  • the drug should specify the intratumoral administration concentration of methylene blue chromosome in its instructions to ensure that it is used as a local active ingredient that can provide local synergistic effects, rather than being mainly used to maximize local effects
  • the chemical ablation agent is applied to avoid risks.
  • the methylene blue dye is preferably selected from the following compounds and their derivatives: methylene blue, patent blue, isosulfur blue, and neomethylene blue.
  • the methylene blue dye is more preferably selected from methylene blue and its derivatives.
  • a local synergistic drug or synergist of the methylene blue dye in the treatment of malignant solid tumors in the treatment of malignant solid tumors it may be an option.
  • the term "nutrient” refers to organic compounds with nutritional and health effects, which are usually used in the preparation of nutritional health products, traditional diets and functional diets (such as health diets), which mainly include amino acid nutrients , Carbohydrate nutrients and lipid nutrients.
  • the concentration of the nutrient in the composition is 2.5-50%, preferably 4-40%.
  • the local synergistic drug or synergist includes amino acid nutrients, and the concentration (w/v) of the amino acid nutrients in the pharmaceutical composition is ⁇ 2%, ⁇ 2.5, ⁇ 5%, preferably ⁇ 7.5%, 10-25% or 18-25%, more preferably 15%-25% or 20%-25%.
  • amino acid nutrients refers to amino acid compounds with nutritional and health effects, preferably selected from amino acids, amino acid polymers and amino acid derivatives with nutritional and health effects, more preferably selected from China , Amino acid nutritional drugs and amino acid excipients with nutritional and health effects contained in the official pharmacopoeias or guidelines of the United States or Europe.
  • amino acid-based nutrients amino acids and amino acid derivatives is preferably a polymer selected from the group of amino acids, or an amino acid of the group of oligopeptide and polypeptide, or the group The amino acid salt in the amino acid: protein amino acid and non-protein amino acid.
  • the protein amino acids include amino acids selected from the following group: non-polar amino acids (such as alanine, valine, leucine, isoleucine, phenylalanine , Proline), polar neutral amino acids (such as tryptophan, tyrosine, serine, cysteine, methionine, asparagine, glutamine, threonine), basic amino acids (such as lysine) Acid, arginine, histidine), acidic amino acids (e.g. aspartic acid, glutamic acid). All the above except glycine are L-type ⁇ -amino acids.
  • non-polar amino acids such as alanine, valine, leucine, isoleucine, phenylalanine , Proline
  • polar neutral amino acids such as tryptophan, tyrosine, serine, cysteine, methionine, asparagine, glutamine, threonine
  • basic amino acids such as lysine
  • Acid arg
  • the non-protein amino acids may include the following amino acids: ⁇ -alanine, taurine, ⁇ -aminobutyric acid (GABA), tea polyphenols (theanine), pumpkin seed amino acids (3-amino-3-carboxypyridine) Alkanoic acid), glutamine, citrulline, ornithine, etc.
  • oligopeptide refers to a polymer of amino acids comprises 2 to 10 identical or different amino acids linked by peptide bonds employed; the term “polypeptide” refers to a same or different 11-100 amino acid polymer linked by peptide bonds.
  • the amino acids constituting the oligopeptide or polypeptide may all be one or more of the above-mentioned amino acids, or may additionally include other amino acids.
  • the oligopeptide may be one or more selected from the group consisting of glycyl-L-tyrosine, glycylalanine, glycylglycine, lysine-glycine di Peptides, glutathione, carnosine ( ⁇ -alanine histidine copolymer), glutathione, collagen oligopeptides, casein hydrolyzed peptides, soybean oligopeptides, oligoarginine, oligoglycine, oligo-lysine Acid.
  • the polypeptide may be one or more selected from the group consisting of polyaspartic acid, polyglutamic acid, and polylysine.
  • amino acid salt refers to the salt formed by the above-mentioned amino acid and acid or base, such as lysine hydrochloride, histidine hydrochloride, glutamic acid hydrochloride , Cysteine hydrochloride, arginine hydrochloride, glycine sulfate, iron glycine sulfate, lysine hydrochloride, aspartic acid hydrochloride, etc.
  • the amino acid nutrient may be one or more of amino acids, amino acid salts, oligopeptides, and polypeptides, for example, 2, 3, 4, or 5 or more. .
  • amino acids, amino acid salts, oligopeptides and polypeptides as the amino acid nutrients are preferably amino acids or their salts selected from the following group, or oligopeptides and polypeptides comprising or consisting of the following amino acids: alanine , Valine, leucine, isoleucine, phenylalanine, proline, tryptophan, tyrosine, serine, cysteine, methionine, threonine, lysine, arginine Acid, histidine, aspartic acid, glutamic acid, ⁇ -alanine, taurine, ⁇ -aminobutyric acid (GABA), theanine, citrulline, ornithine; more preferably selected from Amino acids in the following groups or their salts or oligopeptides and polypeptides comprising or consisting of the following amino acids: arginine, lysine, glycine, cysteine, alanine, serine
  • the amino acid nutrient includes arginine.
  • the amino acid nutrient is selected from compound amino acids including arginine, for example: arginine/tolerant, arginine/serine, arginine/glycine, arginine/cysteine Hydrochloride and so on.
  • the amino acid nutrient includes lysine.
  • the amino acid nutrient includes glycine.
  • the amino acid nutrient includes glutamic acid.
  • the amino acid nutrient is selected from amino acids or amino acid salts with nutritional and health effects, and the concentration (w/v) of the amino acid or amino acid salt in the topical pharmaceutical composition is ⁇ 2%, ⁇ 2.5, ⁇ 5%, ⁇ 7.5%, 10-25% or 18-25%, preferably 15%-25% or 20%-25%.
  • the amino acid nutrients are selected from oligopeptides and polypeptides with nutritional and health effects, and the concentration (w/v) of the oligopeptides and polypeptides in the topical pharmaceutical composition is greater than ⁇ 5%, Preferably it is 7.5-25%, more preferably 10%-25%.
  • the amino acid nutrient is a combination of the amino acid and/or amino acid salt and the oligopeptide and/or polypeptide, and the concentration (w/v) of the combination in the topical pharmaceutical composition is More than ⁇ 5%, preferably 7.5%-25%, more preferably 10-25%.
  • the topical synergistic drug or synergist includes a carbohydrate nutrient, and the concentration (w/v) of the carbohydrate nutrient in the pharmaceutical composition is greater than 5%, preferably ⁇ 10 %, 10-50%, 15-50% or 25-50%.
  • carbohydrate nutrient refers to carbohydrate compounds with nutritional and health effects, preferably selected from monosaccharides, sugar polymers and sugar derivatives with nutritional and health effects, more preferably selected from Carbohydrate nutritional medicines and carbohydrate excipients with nutritional and health effects contained in the official pharmacopoeias or guidelines of China, the United States or Europe.
  • the monosaccharides, sugar polymers, and sugar derivatives as the carbohydrate nutrients are preferably monosaccharides selected from the following groups, sugar polymers containing monosaccharides in the following groups, or their derivatives Substances: glucose, ribose, deoxyribose, xylose, fructose, galactose, fucose.
  • the sugar polymer may be selected from disaccharides, oligosaccharides and polysaccharides containing monosaccharides as described above.
  • disaccharide used refers to a polymer containing two monosaccharides connected by glycosidic bonds
  • oligosaccharide used refers to a polymer containing 3-10 monosaccharides connected by glycosidic bonds.
  • Sugar polymer and the term “polysaccharide” as used refers to a polymer containing more than 10 monosaccharides connected by glycosidic bonds.
  • the monosaccharides constituting the disaccharides, oligosaccharides or polysaccharides may all be one or more of the above-mentioned monosaccharides, or may additionally contain other monosaccharides.
  • the disaccharide may be one or more selected from the group consisting of lactulose, maltose, sucrose, lactose, and trehalose.
  • the oligosaccharide may be one or more selected from the group consisting of chitooligosaccharides, xylo-oligosaccharides, fructooligosaccharides, mannose oligosaccharides, malto-oligosaccharides, and isomalto-oligosaccharides.
  • the polysaccharide may be one or more selected from the group consisting of starch, cellulose, dextran, and glycosaminoglycan.
  • the sugar derivative may be, for example, the following sugar derivatives selected from the above-mentioned monosaccharides or sugar polymers: sugar acid, sugar acid salt, sugar alcohol.
  • sugar acid used refers to acid derivatives of monosaccharides or sugar polymers
  • sugar acid salt refers to salt derivatives of monosaccharides or sugar polymers
  • sugar alcohol refers to monosaccharides or alcohol derivatives of sugar polymers.
  • the sugar acid may be one or more selected from the group consisting of gluconic acid, mannonic acid, and arabinonic acid.
  • the sugar acid salt may be one or more selected from the group consisting of sodium gluconate, sodium mannate, and sodium arabinate.
  • the sugar alcohol may be one or more selected from the group consisting of mannitol, maltitol, lactitol, and xylitol.
  • the carbohydrate nutrient may be one or more of monosaccharides, oligosaccharides, polysaccharides, sugar acids, saccharates, and sugar alcohols, such as 2, 3, 4 Species or 5 species or more.
  • the carbohydrate nutrient is selected from glucose, glucose-containing sugar polymers, or glucose derivatives.
  • the carbohydrate nutrient is selected from ribose, ribose-containing sugar polymers, or ribose derivatives.
  • the carbohydrate nutrient is selected from xylose, xylose-containing sugar polymers, or xylose derivatives.
  • the carbohydrate nutrient is preferably one or more selected from the following: glucose, fructose, chitooligosaccharide, glucosamine, lactulose, sorbitol, ribose, sorbose, mannose, galactose , Sucrose, lactose, trehalose, xylo-oligosaccharides, fructooligosaccharides, mannose oligosaccharides, xylitol, more preferably one or more selected from the following: glucose, sodium gluconate, chitooligosaccharides, glucosamine, Lactulose, ribose, mannose oligosaccharides, xylitol.
  • the concentration (w/v) of the carbohydrate nutrient in the pharmaceutical composition is greater than 5%, preferably ⁇ 10%, 10-40%, 15-50% or 25-50%.
  • the topical synergistic drug or synergist includes lipid nutrients, and the concentration (w/v) of the lipid nutrients in the pharmaceutical composition is ⁇ 4%, preferably 4 -25%.
  • the lipid nutrient includes any pharmaceutically acceptable lipid nutrient, preferably selected from the group of lipids with nutritional and health effects contained in the official pharmacopoeias or guidelines of China, the United States or Europe.
  • the compound is more preferably one or more selected from the group consisting of fats, fatty acids, fat emulsions and lipids.
  • the lipid nutrient is one or more selected from the group consisting of vegetable oil, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), long-chain fat emulsion, Medium chain fat milk, phospholipids.
  • the concentration (w/v) of the lipid nutrient in the pharmaceutical composition is ⁇ 4%, preferably 4-25%.
  • the term "immunomodulator” used means that it has not been used clinically as a vaccine adjuvant and can be used as an immunomodulatory food or immunomodulatory drug, but its molecular structure can be minimized with the preference in the present invention. Instead of maximizing local effects, methylene blue dyes share and provide local synergistic substances. Therefore, the immunomodulator of the present invention is essentially different from the immunoadjuvant in the prior art: 1).
  • the purpose of the composition is different: the purpose of the latter is to promote the relevant and beneficial treatment of tumors in the affected area.
  • the purpose of the former is to provide local synergy; 2).
  • the pharmacological schemes to achieve their goals are different: the latter only involves immune enhancement, while the former must involve chemical enhancement (local synergy); 3).
  • the components (structures) of the (efficiency) scheme are different: the latter is selected from one or more of the following groups commonly used as vaccine adjuvants: inorganic adjuvants, (highly immunogenic) microorganisms and their products, Synthetic adjuvants, oils, cytokines, heterologous antigens, and the former is selected from probiotic components or/and immunomodulatory peptides that minimize the immunogenicity of bacteria that are usually not used as vaccine adjuvants but used as immunomodulatory foods, and usually Immunoglobulin used as an immunomodulator of a vaccine adjuvant that is completely different from the antigen-enhancing effect; 4).
  • the latter s technical effect is mainly beneficial to the immune enhancement of anti-tumor antigens, but Usually there is no tissue destruction effect, and the necessary technical effect of the former is a local synergy that can cause effective tissue destruction, which is mainly a chemical effect that does not depend on immune action. In the following examples, the realization of this local synergy does not depend on the immune function of the animal, and it can even be achieved in nude mice with severe immunodeficiency.
  • probiotics refers to non-pathogenic living microorganisms that can have a beneficial effect on the health of the host.
  • Probiotics include cells, and yeasts are single-celled microorganisms.
  • probiotic components refers to preparations derived from natural probiotics or engineered bacteria (such as wall polysaccharides) or engineered analogs of the preparations (such as synthetic polysaccharides similar to cell wall polysaccharides or other Source polysaccharide).
  • the probiotic components are preferably selected from those that minimize bacterial immunogenicity, and more preferably selected from one or more of the following groups and their engineering analogs: water-soluble components of probiotics , Probiotics components water-insoluble components, inactivated probiotics.
  • bacterial immunogenicity refers to the ability of bacteria as a complete foreign object to produce an immune response in the recipient's body, and different bacteria have different bacterial immunogenicity. Live probiotics have the strongest bacterial immunogenicity, but direct entry into the body also has a strong safety risk.
  • inactivated probiotic bacteria refers to a preparation obtained after a bacterial inactivation engineering process, wherein the bacterial inactivation is, for example, one or more of the following: high temperature inactivation, high temperature and high pressure inactivation, ultraviolet inactivation, chemical Reagent inactivation, radiation inactivation.
  • probiotic water-insoluble component refers to any component obtained from probiotics with a water solubility of ⁇ 0.1% (such as broken probiotic precipitates, water-insoluble probiotic cell wall components, water-insoluble polysaccharides, water-insoluble ⁇ - Dextran, etc.).
  • water-soluble components of probiotics refers to any components obtained from probiotics with a water solubility of ⁇ 0.1% (for example, water-soluble extracts of probiotics, water-soluble polysaccharides, water-soluble ⁇ -glucans, etc. ).
  • the administration concentration of the inactivated probiotic in the pharmaceutical composition is >0.3%, ⁇ 0.75%, 0.75-15 , Preferably 1.5-15% or 5-15%; when the probiotic component includes the probiotic water-soluble component, the administration concentration of the probiotic water-soluble component in the pharmaceutical composition is >0.25, or 0.35-25%, preferably 0.35-15% or 5-15%; when the probiotic component includes the probiotic water-insoluble component, the probiotic water-insoluble component particles are The administration concentration in the pharmaceutical composition is >0.5, or 0.5-15%, preferably 1.5-15% or 5-15%.
  • the probiotic bacteria are selected from one or more of the following groups of natural bacteria or engineered bacteria: probiotic bacillus, probiotic lactic acid bacteria, probiotic bifidobacteria, and probiotic fungi.
  • the probiotic bacteria include one or more selected from probiotic Bacillus. In one embodiment, the probiotic bacteria include one or more selected from probiotic Lactobacillus. In one embodiment, the probiotic bacteria include one or more selected from probiotic bifidobacteria. In one embodiment, the probiotics include one or more selected from probiotic fungi.
  • the Bacillus is selected from one or more of the following: Bacillus cereus, Bacillus licheniformis, Bacillus subtilis, Bacillus megaterium, Bacillus firmus, Bacillus coagulans Bacillus, Bacillus lentus, Bacillus pumilus, Bacillus natto. In one embodiment, the Bacillus is preferably selected from one or more of the following: Bacillus licheniformis, Bacillus subtilis, Bacillus pumilus.
  • the lactic acid bacteria include one or more selected from the group consisting of Lactobacillus or/and Bifidobacterium.
  • the Lactobacillus includes one or more selected from the group consisting of Lactobacillus acidophilus, Lactobacillus salivarius, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus pumilus, and Lactobacillus fermentum.
  • the Lactobacillus is preferably selected from one or more of the following: Lactobacillus casei, Lactobacillus plantarum, Lactobacillus pumilus, and Lactobacillus fermentum.
  • the bifidobacterium includes one or more selected from the group consisting of: Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium breve, Bifidobacterium infantis, Lactobacillus brucelli, Swiss Lactobacillus, Lactobacillus thermophilus, Enterococcus faecium, Streptococcus faecalis.
  • the bifidobacterium is preferably selected from one or more of the following: Bifidobacterium longum, Bifidobacterium adolescentis, Enterococcus faecium, and Streptococcus faecalis.
  • the fungus includes one or more selected from yeast or/and Brucella saccharomyces, wherein the yeast includes one or more selected from the group consisting of Saccharomyces cerevisiae, Delta Saccharomyces cerevisiae, Candida, Wickham's yeast, Pichia pastoris, Saccharomyces leucocephalus, Saccharomyces sp , Candida Ruan.
  • the probiotic is preferably one or more selected from yeast or/and Brucella.
  • the probiotics are preferably selected from probiotics that do not contain mycolic acid cell walls.
  • the probiotic is preferably selected from Saccharomyces brucei.
  • the probiotic is preferably selected from Saccharomyces cerevisiae.
  • immunoglobulin is a structural concept that refers to all drugs that are the same or similar to antibody molecules, including natural immunoglobulins and their engineering analogs.
  • the natural immunoglobulins include immunoglobulin G, immunoglobulin A, immunoglobulin M, immunoglobulin D, and immunoglobulin E, which are usually obtained from the separation of animal plasma, such as intravenous immunoglobulin G and intravenous injection. Injection of immunoglobulin A, intravenous injection of immunoglobulin M, and so on.
  • the engineering analogue of natural immunoglobulin refers to an engineering product (including genetic engineering product) that is different from natural immunoglobulin but has all or part of the key structure of natural immunoglobulin, such as immunoglobulin complex, Immunoglobulin Fc fragments, immunoglobulin Fab fragments, immunoglobulin Fab' fragments, immunoglobulin F(ab') fragments, single immunoglobulins, etc.
  • the administration concentration (w/v) of the immunoglobulin in the pharmaceutical composition is >1.5%, ⁇ 2.0, 2.0-35%, preferably 3.0-30%.
  • the term "immunomodulatory peptide” refers to a non-protein polypeptide derived from organisms or food proteins that has an immunomodulatory effect, for example, the CAS number used in the examples of the present invention is 63231-63-0 Ribonucleic acid.
  • the administration concentration (w/v) of the immunomodulatory peptide in the pharmaceutical composition is >1.5%, ⁇ 2.0, 2.0-35%, preferably 3.0-30%.
  • conventional antineoplastic drug refers to a drug that can effectively inhibit solid tumors by absorption at a safe dose, which is selected from any pharmaceutically acceptable conventional antineoplastic drugs, preferably selected from the present invention.
  • Conventional anti-tumor drugs known in the art are more preferably selected from China, the United States or Europe, which has been approved or will be approved by the official competent administrative department of China, the United States or Europe (such as the FDA or China Food and Drug Administration), or has been included in the official pharmacopoeia of China, the United States or Europe. The anti-tumor drugs to be loaded.
  • absorption refers to the pharmacological effect of the drug being absorbed by the blood to form the drug-carrying blood into the target area.
  • Some conventional anti-tumor drugs have been expected to improve their efficacy through intratumoral administration, but the increase in drug efficacy with the increase in concentration (chemical kinetics) is far lower than its theoretical correlation.
  • different drugs were shared locally in a large number of studies, there was not much local synergy, let alone unexpected synergy, indicating that the synergy of these drugs under special conditions in the tumor (such as the microenvironment of cancer cells) has a high degree of inefficiency. Certainty. .
  • the conventional anti-tumor drugs may be one or more selected from the group consisting of: drugs that disrupt the structure and function of DNA, drugs that intercalate in DNA and interfere with transcription of RNA, drugs that interfere with DNA synthesis, and proteins that affect the structure and function of DNA. Synthetic drugs.
  • the drugs that disrupt DNA structure and function include, for example, alkylating agents (e.g., cyclophosphamide, carmustine, etc.), metal platinum complexes (e.g., cisplatin, carboplatin, etc.), DNA topoisomerase inhibitors ( For example, doxorubicin, topotecan, irinotecan, etc.).
  • the drugs that intercalate DNA to interfere with transcription of RNA include, for example, anti-tumor antibiotics, such as actinomycins, daunorubicin, doxorubicin, and the like.
  • the drugs that interfere with DNA synthesis include, for example, pyrimidine antagonists (such as uracil derivatives 5-fluorouracil, furfurouracil, difurfurouracil, cytosine derivatives cytarabine, cyclocytidine, 5-azacytidine, etc.) , Purine antagonists (e.g. oncolytic, thioguanine, etc.), folic acid antagonists (e.g., methotrexate, etc.).
  • the drugs that affect protein synthesis include, for example, colchicines, vinblastines, taxanes (such as paclitaxel, docetaxel, etc.) and the like.
  • the pharmaceutically acceptable liquid carrier includes water and/or ethanol.
  • the pharmacologically acceptable liquid carrier is mainly selected according to the properties of conventional anti-tumor drugs, so that the drug can reach a corresponding concentration.
  • the conventional anti-tumor drug is selected from the group consisting of water-soluble conventional anti-tumor drugs and alcohol-soluble conventional anti-tumor drugs.
  • the term "alcohol-soluble conventional anti-tumor drugs” refers to conventional anti-tumor drugs whose solubility in ethanol or ethanol aqueous solution at room temperature is greater than or equal to the concentration required for effective local action, which includes, for example, yew Alkanes, vinblastines, etc.
  • water-soluble conventional antitumor drug refers to a conventional antitumor drug whose solubility in aqueous solution at room temperature is greater than or equal to the concentration required for its effective local action, which includes, for example, one or more water-soluble drugs selected from the following groups Sexual compounds: uracil derivatives, cyclophosphamide, gemcitabine (such as gemcitabine hydrochloride), epirubicin (such as epirubicin hydrochloride), antitumor antibiotics (such as doxorubicin, actinomycetes) Vinblastine, etc.), vinblastines (for example, vinblastine sulfate), teniposide, metal platinum complexes, etc.
  • sexual compounds uracil derivatives, cyclophosphamide, gemcitabine (such as gemcitabine hydrochloride), epirubicin (such as epirubicin hydrochloride), antitumor antibiotics (such as doxorubicin, actinomycetes) Vinblastine, etc
  • the conventional anti-tumor drugs may be one or more selected from the following groups: uracil derivatives, cyclophosphamides, gemcitabine, epirubicin, Anti-tumor antibiotics, teniposide, metal platinum complexes, taxanes; preferably one or more selected from the following drugs and their analogous derivatives: 5-fluorouracil, cyclophosphamide, gemcitabine, epirubin Bicin, antitumor antibiotic, teniposide, metal platinum complex, paclitaxel.
  • the concentration of the conventional anti-tumor drug is greater than 30% of its saturated concentration, preferably 50-100%, 60-100%, 70-100%, 80% of its saturated concentration. -100%, or 90-100%, wherein the saturated concentration refers to the saturated concentration of the conventional anti-tumor drug in the liquid carrier.
  • the concentration (w/v) of a conventional anti-tumor drug (such as cyclophosphamide, carmustine, etc.) selected from the alkylating agent in the topical pharmaceutical composition is 0.5-6 %, preferably 0.75-1.5%.
  • the concentration (w/v) of a conventional anti-tumor drug (such as cisplatin, carboplatin, etc.) selected from the metal platinum complex in the topical pharmaceutical composition is 0.03-0.08% , Preferably 0.03-0.06%.
  • the concentration of a conventional anti-tumor drug (such as doxorubicin, topotecan, irinotecan, etc.) selected from the DNA topoisomerase inhibitor in the topical pharmaceutical composition is 0.05-0.20%, preferably 0.75-0.15%.
  • the concentration (w/v) of conventional anti-tumor drugs selected from the anti-tumor antibiotics (such as actinomycins, daunorubicin, etc.) in the topical pharmaceutical composition is 1 -4%, preferably 1-2%.
  • a conventional antitumor drug selected from the pyrimidine antagonist for example, uracil derivative 5-fluorouracil, furfurouracil, difurfurouracil, cytosine derivative cytarabine, cyclocytidine, 5
  • concentration (w/v) of azacytidine, etc.) in the topical pharmaceutical composition is 0.5-2%, preferably 0.75-1.5%.
  • the concentration (w/v) of a conventional anti-tumor drug selected from the taxanes (such as paclitaxel, docetaxel, etc.) in the topical pharmaceutical composition is 0.5-2% , Preferably 0.75-1.5%.
  • composition disclosed according to the present application it also optionally includes one or more selected from the group consisting of analgesics, sustained-release carriers, pH adjusters, and excipients.
  • the pharmaceutical composition disclosed according to the present application may further optionally include an analgesic.
  • the analgesic is used to alleviate the pain of the patient, and it may be any suitable one known to those skilled in the art, such as benzyl alcohol, procaine hydrochloride, chlorobutanol, lidocaine hydrochloride and the like.
  • the concentration of the analgesic in the pharmaceutical composition may be, for example, 0.1-4% by weight.
  • the concentration of benzyl alcohol in the pharmaceutical composition may be 1-4% by weight
  • the concentration of procaine hydrochloride, chlorobutanol, and lidoca hydrochloride in the pharmaceutical composition may be 1- 3% by weight.
  • the pharmaceutical composition disclosed according to the present application may further optionally include a sustained-release carrier.
  • the sustained-release carrier may be any suitable one known to those skilled in the art, including, for example, a gel matrix, a particulate carrier, a micellar matrix, and the like.
  • the concentration (w/v) of the sustained-release carrier in the pharmaceutical composition can be, for example, 0.5-13%, preferably 1-12% or 1-15%.
  • the pharmaceutical composition disclosed according to the present application may further optionally include excipients.
  • the excipient may be any suitable one known to those skilled in the art, which may include, for example, one or more of the following: dispersion medium, preservative, stabilizer, wetting agent and/or emulsifier, solubilizer, Tackifiers, etc.
  • the viscosity increasing agent is, for example, sodium carboxymethyl cellulose, carboxymethyl cellulose, polyvinylpyrrolidone or gelatin.
  • the preservatives are, for example, antioxidants (such as ascorbic acid).
  • the pharmaceutical composition disclosed according to the present application may include active ingredients (the methylene blue dye, the topical synergistic drug or synergist, and optionally other active ingredients as described above) and a liquid carrier (such as Water, ethanol, or water/ethanol mixture) any dosage form suitable for intratumoral administration, preferably the following dosage forms: injections (preferably local injections), external liquids, nebulizers, and the like.
  • active ingredients the methylene blue dye, the topical synergistic drug or synergist, and optionally other active ingredients as described above
  • a liquid carrier such as Water, ethanol, or water/ethanol mixture
  • any dosage form suitable for intratumoral administration preferably the following dosage forms: injections (preferably local injections), external liquids, nebulizers, and the like.
  • injection refers to a sterile preparation containing an active ingredient and a liquid carrier for in vivo administration.
  • the injections are divided into local injections, intravenous injections, etc. according to the mode of administration, and intravenous injections can be used as local injections only after a given intratumoral administration concentration.
  • Injections are classified into liquid injections, powder injections for injection, etc. according to their commercial form.
  • the injection powder contains sterile dry powder and a solvent, the sterile dry powder contains part or all of the active ingredients, and the solvent contains all the liquid carriers.
  • the concentration of the active ingredient in the injection is the concentration of the active ingredient in the mixture with all the liquid carriers, which is usually the end point of the intratumoral drug delivery device (syringe, puncture, injection catheter, etc.) (such as needle hole, catheter outlet, etc.) Etc.)
  • the concentration of the active ingredient in the liquid medicine is usually the end point of the intratumoral drug delivery device (syringe, puncture, injection catheter, etc.) (such as needle hole, catheter outlet, etc.) Etc.)
  • the concentration of the active ingredient in the liquid medicine is usually the end point of the intratumoral drug delivery device (syringe, puncture, injection catheter, etc.) (such as needle hole, catheter outlet, etc.) Etc.)
  • the concentration of the active ingredient in the liquid medicine is usually the end point of the intratumoral drug delivery device (syringe, puncture, injection catheter, etc.) (such as needle hole, catheter outlet, etc.) Etc.)
  • the concentration of the active ingredient in the liquid medicine is usually the end
  • a topical pharmaceutical composition for the treatment of malignant solid tumors in a freeze-dried or semi-lyophilized form which is lyophilized or semi-lyophilized according to the disclosed Part or all of the methylene blue dye, the local synergistic drug or synergist, and the pharmaceutical composition of a pharmaceutically acceptable carrier are obtained.
  • composition of the present invention should be made into a dosage form that can be administered into the target area intratumorally, preferably a topical pharmaceutical dosage form.
  • the preparation of the pharmaceutical composition of the present invention includes the following steps: preparing a liquid medicine containing the methylene blue dye, nutrients and/or conventional anti-tumor, liquid medium and optionally other substances.
  • the liquid medicine may be a solution (for example, a solution in a hydrophilic vehicle, preferably an aqueous solution), a suspension, or an emulsion containing a topical active ingredient.
  • the dispersion medium therein can be any suitable one known to those skilled in the art, such as micro-materials or nano-materials.
  • the dispersion medium therein can be any suitable one known to those skilled in the art, such as vegetable oil, synthetic oil or semi-synthetic oil that can be used for injection.
  • the vegetable oil may be, for example, cottonseed oil, almond oil, olive oil, castor oil, sesame oil, soybean oil, and peanut oil.
  • the pharmaceutical composition liquid injection of the present invention can be prepared by a method comprising the following steps: 1) The necessary components (such as the methylene blue Dyestuffs, the local synergistic drug or synergist) and optional other components are added to the vehicle to prepare a liquid; 2) other necessary components (such as other nutrients) in the required amount according to the intratumoral administration concentration and Optionally, add the liquid prepared in 1) and mix uniformly to obtain a liquid medicine; 3) sterilize the liquid medicine prepared in 2) and prepare a liquid injection.
  • the sterilizing liquid medicine in the liquid injection can be used directly or diluted as a liquid medicine for intratumoral administration.
  • the pharmaceutical composition liquid injection of the present invention can be prepared by a method including the following steps: 1) The required amount of methylene blue dye and the local synergistic drug are added according to the intratumoral administration concentration. Or the synergist and optional other components are added to the solvent (or pharmaceutically acceptable liquid carrier) and mixed uniformly and prepared as a sterile liquid I after sterilization; 2) The required amount will be based on the intratumoral administration concentration.
  • the optional other components (such as acidulant) are added to the solvent (or pharmaceutically acceptable liquid carrier), mixed uniformly, and sterilized to prepare the sterilizing liquid II.
  • the sterilizing liquid I and the sterilizing liquid II form a mixed liquid before or after entering the intratumoral administration device, which can be used directly or diluted as a liquid drug for intratumoral administration.
  • the injection powder of the pharmaceutical composition of the present invention can be prepared by a method including the following steps: The sterile dry powder of the local synergistic drug or synergist; and the preparation of a sterile vehicle containing the required amount of the other components (such as amino acid nutrients, analgesics, etc.) according to the intratumor administration concentration.
  • the sterile dry powder is preferably a sterile freeze-dried dry powder, and its preparation method includes: 1) preparing a solution containing methylene blue dye, the local synergistic drug or synergist and optional other components; 2) removing Bacteria filtration and packaging; 3) freeze-drying; 4) plugging and capping.
  • the process conditions of the freeze-drying include, for example, the pre-freezing condition is kept at the pre-freezing temperature -45°C for 4 hours; the sublimation drying condition is that the temperature rise rate is 0.1°C/min and the temperature is raised to -15°C and kept for at least 10 hours; The adsorption drying conditions are 30°C for 6 hours.
  • the sterile dry powder of the injection powder is re-dissolved in a sterile solvent to form a re-solution drug, which can be used directly or diluted as a liquid drug for intratumoral administration.
  • the changes in the composition of the present invention include: containing different types and concentrations of the methylene blue dye, containing different types and concentrations of the local synergistic drugs or synergists, containing different types and concentrations of other additives ( For example, analgesics, acidifiers, etc.).
  • the pharmaceutical composition is mainly used to treat malignant solid tumors, especially refractory malignant solid tumors (such as pancreatic cancer) by intratumoral administration.
  • Intratumoral administration requires that the composition of the drug (local active ingredients, composition ratio and component concentration) can be administered into the tumor by interventional means, and produce the desired therapeutic effect in it.
  • the composition of the drug local active ingredients, composition ratio and component concentration
  • the treatment of malignant solid tumors by the composition of the present invention preferably includes tumor treatment.
  • the composition of the present invention for tumor treatment of malignant solid tumors preferably includes effective destruction of non-malignant tumor cell components in the tumor or tumor-related nodules, especially connective tissue.
  • treatment refers to any reasonable and advantageous medical treatment, which includes post-diagnosis treatment (for example, medical treatment performed after diagnosis) and preventive treatment (for example, medical treatment that can be performed without a diagnosis) .
  • treatment of malignant solid tumors or “treatment of malignant solid tumors” refers to the treatment of patients who have been diagnosed with or are likely to have malignant solid tumors, including tumor treatment and other tissues containing tumor cells (such as lymphatic tissue) the treatment.
  • tumor treatment refers to the treatment of tumors or tumor-related nodules.
  • the preferred solution for the indications of the composition of the present invention is one or more of the following: a larger stromal ratio tumor body that can be administered locally, and the treatment of tumors containing the tumor body ; Treatment of larger tumors that can be locally administered and tumors containing the tumors; treatment of malignant tumor-related nodules that can be administered locally.
  • tumor body refers to a mass formed due to abnormal proliferation of cells or mutated cells.
  • solid tumor (sometimes abbreviated as tumor) refers to a disease in which tumors can appear.
  • malignant solid tumor (sometimes abbreviated as malignant tumor) refers to a malignant disease in which tumors can appear.
  • Tumors include, for example, the following groups classified according to tumor cell types: epithelial cell tumors, sarcomas, lymphomas, germ cell tumors, and blastoma; and include tumors named according to the organ or tissue where the tumor cell concentration area is located, including, for example, according to Tumors named for the following organs or tissues: skin, bone, muscle, breast, kidney, liver, lung, gallbladder, pancreas, brain, esophagus, bladder muscle, large intestine, small intestine, spleen, stomach, prostate, testis, ovary or uterus.
  • the malignant solid tumors include, for example, breast cancer, pancreatic cancer, thyroid cancer, nasopharyngeal cancer, prostate cancer, liver cancer, lung cancer, bowel cancer, oral cancer, esophageal cancer, stomach cancer, laryngeal cancer, testicular cancer, and vaginal cancer. , Uterine cancer, ovarian cancer, etc.
  • the local drug in the present invention is a therapeutic drug.
  • it can also be combined with other interventional therapies, systemic chemotherapy, immunotherapy, photodynamic therapy, sonodynamic therapy, surgical intervention or Combinations of such therapies are administered in combination to further improve the efficacy.
  • the pharmaceutical composition is mainly used for the treatment of malignant solid tumors by intratumoral administration.
  • the acidifying agent and the ineffective absorption compound are administered intratumorally at their concentration or amount in the local pharmaceutical composition. This concentration or amount can provide a synergistic effect of local response compared to intratumoral administration.
  • composition of the present invention has been shown to promote the effective destruction of non-malignant tumor cell components (especially connective tissue) in tumors or tumor-related nodules. At the same time, there is only minimal damage to the patient's normal tissues, so as to achieve safe and effective treatment of malignant solid tumors.
  • the immunomodulators used in the following specific examples can all be obtained from commercial sources.
  • the strains of probiotics used are all strains used in Saccharomyces cerevisiae and probiotic preparations approved by the food and drug authorities or contained in the official pharmacopoeia.
  • strains contained in probiotic preparations approved by the Chinese pharmaceutical authority Clostridium butyricum in Bacillus coagulans live tablets, Bacillus licheniformis strains in live Bacillus licheniformis granules (capsules), Oral Bacillus cereus Bacillus cereus strains in live bacteria preparations, Clostridium butyricum strains in live Clostridium butyricum capsules, Saccharomyces boulardii strains in Saccharomyces boulardii powder (capsules), Saccharomyces boulardii strains in Lactobacillus acidophilus capsules Lactobacillus butyricum strains, Clostridium butyricum strains in oral clostridium butyricum viable bacteria powders, Enterococcus faecium strains, subtilis strains in Bacillus subtilis double live bacteria multi-dimensional granules, and Enterococcus butyricum triple viable bacteria tablets Lactic acid strain
  • L-amino acids are abbreviated as amino acids (for example, L-arginine is abbreviated as arginine), reduced glutathione is abbreviated as glutathione, and alanyl-glutamine dipeptide is abbreviated as For glutathione.
  • the animal tests of subcutaneous transplantation tumors are carried out in accordance with the test guidelines issued by the drug administration authority.
  • the test animals are Balb/c nude mice or mice aged 6-8 weeks and weighing 17.5-20.5 g.
  • the subcutaneous transplantation is carried out according to the conventional method of subcutaneous inoculation of tumor cells.
  • the PEMS 3.2 software compiled by West China School of Public Health, Sichuan University
  • Test observation, measurement and analysis items including general status, body weight, food intake, tumor volume, tumor weight, etc.
  • Tumor volume V 1/2 ⁇ a ⁇ b 2 , where a represents the length of the tumor and b represents the width of the tumor.
  • Tumor inhibition rate R(%) (CW-TW)/CW ⁇ 100%, where TW is the average tumor weight of the study group; CW is the average tumor weight of the negative control group.
  • E i i the drug efficacy referred to as E i i, where E i may be a (100-r i)% or R i%.
  • B/A the combination of A drug and B drug
  • the single-use drug effects of A and B are recorded as E A and E B respectively
  • the actual common drug effects of A/B are recorded as E A+B .
  • test results (such as tumor weight, tumor volume) are expressed as mean ⁇ standard deviation (x ⁇ s), and the difference between the two test animal groups and the group mean uses the statistical software SPSS 13.0 or SPSS The 19.0 software performs a significance test for comparison.
  • the test uses the statistic t to perform the test.
  • the type of drug action (shared pharmacology) is studied through drug effects, especially comparing the drug effects of the same research drug in different technical solutions.
  • the difference in drug effect of drug i between regimens X and Y is not unusually large (for example, E iX /E iY ⁇ 150%), it is likely to be basically the same drug action type (pharmacology) with different dynamics Due to scientific conditions (action concentration); and when the effect of the drug is unusually large (for example, E iX /E iY >150%), the drug effect of drug i in plan X should exceed its effect in plan X
  • the kinetic expected range of the drug action type (pharmacology) in Y which is likely to involve a drug action type (pharmacology) different from that of the scheme Y.
  • the positive controls for chemotherapy include classic cytotoxic drugs (e.g. 0.5-1% 5-fluorouracil, which has a tumor inhibition rate of ⁇ 30% under the conditions of the following examples) and classic chemical ablation agents (e.g. 75- 99% ethanol, its tumor inhibition rate under local administration conditions in the following examples is ⁇ 30%).
  • classic cytotoxic drugs e.g. 0.5-1% 5-fluorouracil, which has a tumor inhibition rate of ⁇ 30% under the conditions of the following examples
  • classic chemical ablation agents e.g. 75- 99% ethanol, its tumor inhibition rate under local administration conditions in the following examples is ⁇ 30%).
  • E A/B is the actual common drug effect of composition A/B (such as tumor inhibition rate)
  • E A and E B are the actual single-drug efficacy of component A and component B of composition A/B, respectively
  • (E A + E B- E A ⁇ E B ) is the expected effect of component A and component B based on the theory of the actual efficacy of the single drug.
  • the actual/expected ratio q is calculated according to the q calculation formula in the above-mentioned Jin Zhengjun's modified method, and the common effect of the combined administration according to the above-mentioned Zhang Xiaowen-Jin Zhengjun method is judged as follows:
  • composition A/B group does not show therapeutically significant pharmacological effects (for example, the tumor inhibition rate is less than 30%), the combined administration of A and B does not show a therapeutically meaningful shared effect, which is regarded as No synergy.
  • the composition group shows therapeutic effect (for example, tumor inhibition rate ⁇ 30%)
  • the actual/expected ratio q is between 0.85 and 1.15
  • the common effect of the composition is additive (the actual effect is in line with the theoretical Simply add up expectations); if the actual/expected ratio q>1.15, the sharing effect of the composition is obviously synergistic (the actual effect exceeds the theoretical simple addition expected); if the actual/expected ratio q ⁇ 0.85, the sharing of the composition
  • the effect is an obvious antagonistic effect (the actual effect is not as good as the theoretical simple addition expectation).
  • composition of part of the composition (X%/Y%) of the present invention prepared in this example is listed in the following table.
  • Test 1A Preparation of liquid injection (1)
  • methylene blue dye e.g. 1g of methylene blue
  • local synergists e.g. 10g of glutathione
  • optional other components e.g. dilute to the total volume according to the required concentration (as described in Table 2)
  • the volume (e.g. 100ml) of liquid carrier e.g. water for injection
  • the preparation for example, 1% methylene blue/10% glutathione in water
  • Solution I and solution II are mixed evenly according to the required concentration of each component (for example, 8.5ml of solution I and 1.5ml of solution II are mixed) into a mixed solution (for example, 1% methylene blue/30% glucose/5% acetic acid aqueous solution) )
  • a mixed solution for example, 1% methylene blue/30% glucose/5% acetic acid aqueous solution
  • Test 1C Preparation of powder injection for injection
  • methylene blue dye e.g. 1g methylene blue
  • local synergists e.g. 20g arginine, 1g 5-fluorouracil
  • optionally other groups according to the required concentration (as described in Table 2)
  • Divide and dilute to the total volume (e.g. 100ml) of the liquid carrier e.g. water for injection
  • the liquid carrier e.g. water for injection
  • %Methylene blue/20%arginine/5-fluorouracil aqueous solution can be used as a liquid drug for intratumoral administration.
  • Test 2A The test animal was a mouse, the modeling cell was a sarcoma S180 cell, and the transplanted tumor was modeled under the skin of the animal's right axillary at 1 ⁇ 10 6 cells/only.
  • Successfully modeled experimental animals (mice bearing malignant sarcoma, with an average tumor volume of 121mm 3 ) were randomly divided into 2 series (series I and II), and each series was divided into a negative control group (I0 and II0) And 11 study groups (I1-I11 and II1-II11).
  • the negative control is normal saline, and the 11 study drugs are shown in the table below, series I for systemic administration (intraperitoneal injection) and series II for local administration (intratumoral injection).
  • the medicines are all aqueous solutions, which are prepared according to the preparation method of Example 1.
  • the medicine is administered once every 3 days, a total of 3 times, and the injection volume is less than or equal to 120 ⁇ l/mouse.
  • the animals were euthanized 10 days after the end of the medication, and the tumor weight (W) was measured after anatomy, and the tumor inhibition rate (R) was calculated from the negative control group of each medication mode. The results are shown in the following table.
  • the intraperitoneal injection group (group I1) and the intratumoral injection group (group II-1) of the positive control substance (5-fluorouracil) showed almost indistinguishable tumor inhibition rates, although some people believe that the intratumoral drug can be used.
  • Increasing its local drug concentration will significantly improve its efficacy.
  • This result shows that the drug did not substantially change its targeting (tumor cells) and pharmacology (inhibition of tumor cells) when injected intratumorally. Therefore, unless placed in a sustained-release system, conventional anti-tumor drugs are still mainly administered systemically rather than locally.
  • the methylene blue used is lower than the concentration of its clinical injection ( ⁇ 1%), and it is considered to be methylene blue that does not produce adverse local effects even if injected intravenously.
  • the difference between the tumor weight of the study group I2 and the negative control group I0 is not statistically significant (P>0.05), and its tumor inhibition rate is at a negligibly low level, indicating that the methylene blue dye
  • the conventional anti-tumor effect of serotonin can not produce tumor growth inhibitory effect.
  • the results of study group II2 are consistent with tumor study group I2, indicating that the methylene blue dye only provides a minimized rather than maximized local effect.
  • each methylene blue dye/nutrient composition group (6-8) in series I is exactly the same as that of methylene blue dye single-drug group 5 and the corresponding nutrient single-drug group (1-4) in the same series. It shows that the conventional combination of the methylene blue dye and the common substance (nutrient) (whether it is a combination of nutrition and conventional anti-tumor effects, or a combination between conventional anti-tumor effects) can not produce tumor growth inhibitory effects, more It is not an effective synergy.
  • the composition group 9 in the series I did not increase the tumor inhibition rate, indicating that any conventional combination of methylene blue dye and conventional anti-tumor drugs in the conventional composition (such as conventional anti-tumor effects) The combination) can not achieve tumor suppression and synergy.
  • the tumor inhibition rate of group II9 is more than 150% higher than that of group I9, indicating that the topical pharmaceutical composition in series II achieves effective tumor treatment through a new pharmacology that is different from the conventional pharmaceutical composition in series I.
  • the tumor inhibition rate of the composition group 9 is also beyond the expected range of the purely additive effect of the two drugs, and its actual/expected ratio (q) is greater than 1.15, showing The above-mentioned new pharmacology should be the local synergistic pharmacology.
  • the tumor inhibition rate of group II11 is more than 150% higher than that of group I11, indicating that the topical pharmaceutical composition in series II achieves effective tumor treatment through a new pharmacology that is different from the conventional pharmaceutical composition in series I.
  • the tumor inhibition rate of the composition group 11 is also beyond the expected range of the theoretical simple additive effect of the two drugs, and its actual/expected ratio (q) is greater than 1.15, showing The above-mentioned new pharmacology should be the local synergistic pharmacology.
  • Test 2B The test animal was a nude mouse, the modeling cell was human pancreatic cancer cell (PANC-1), and the transplanted tumor was modeled at 1 ⁇ 10 6 cells/only under the skin of the animal's right axillary.
  • Successfully modeled test animals (nude mice bearing pancreatic cancer, with an average tumor volume of 147mm 3 ) were randomly divided into 2 series (series I and II), and each series was divided into a negative control group (I0 and II0) And 11 study groups (I1-I11 and II1-II11).
  • the negative control is physiological saline, and the 11 study drugs are shown in the following table.
  • immunoglobulin is human immunoglobulin
  • probiotics are heat-inactivated Saccharomyces boulardii
  • the water-soluble components of probiotics are water-soluble yeast glucosamine.
  • the sugar, water-insoluble component of probiotics is Saccharomyces cerevisiae ⁇ -glucan.
  • the medicines are all liquids and are prepared according to the preparation method of Example 1. Series I line systemic administration (intraperitoneal injection) and series II line local administration (intratumoral injection). In the present invention, all intratumoral injections need to avoid blood vessels.
  • the medicine is administered once every 3 days, a total of 3 times, and the injection volume is 120 ⁇ l/mouse.
  • Ten days after the end of the medication the animals were euthanized, and the tumor weight (W) was measured after anatomy, and the tumor inhibition rate (R) was calculated from the negative control group of each medication method. The results are shown in the following table.
  • the active combination (referred to as conventional combination in the present invention) that can be provided by drug sharing is usually studied through systemic administration like the series I in the above two experiments.
  • the mainstream view is that local administration is just another mode of administration that is different from systemic administration. It may have an increase in kinetic concentration but no increase in technical content, and it will not be expected to improve the kinetics of conventional combination pharmacology.
  • new pharmacology is generated.
  • Methylene blue is mainly used clinically as an antidote or vital dye.
  • it also has properties similar to ethanol: its maximum cytotoxic effect can be used in liquid reactors (cell test or non-solid tumor animal test), but not in non-liquid reactors (animal intratumoral administration test) Shows effective anti-tumor efficacy, and its maximum local effect can show effective anti-tumor efficacy in non-liquid reactors, but not in liquid reactors.
  • the combination (X/Y) of methylene blue (X) and any drug (Y) with minimal local effects can only produce conventional combinations (such as X conventional anti-tumor effects and X/Y) when administered systemically.
  • the combination of Y's immunomodulatory effect, or/and the combination of Y's conventional anti-tumor effect), and X/Y can only show its function in systemic medication when administered locally.
  • the study group 11 in the series I did not have an increase in tumor inhibition rate compared with 1, indicating that the methylene blue dye and the complex (immunomodulator/conventional anti-tumor drug) are any conventional combination in the conventional composition (for example, the combination of immunomodulatory effect and conventional anti-tumor effect, or/and the combination between conventional anti-tumor effect) can not achieve tumor suppression synergistic effect, let alone effective synergistic effect.
  • the tumor inhibition rate of group II11 is more than 150% higher than that of group I11, indicating that the topical pharmaceutical composition in series II achieves effective tumor treatment through a new pharmacology that is different from the conventional pharmaceutical composition in series I.
  • the tumor inhibition rate of the composition group 11 is also beyond the expected range of the purely additive effect of the two drugs, and its actual/expected ratio (q) is greater than 1.15, indicating that the above-mentioned new pharmacological principle should be The local synergistic pharmacology.
  • Drugs are not substances (X) completely defined by chemistry, but active substances (AX) with uniform structure (X) and effect (A).
  • AX active substances
  • different substances (X1 and X2, such as methylene blue dyes and conventional anti-tumor drugs in the above systemic administration series) that provide the same activity (A, such as cytotoxicity) can easily be regarded as different drugs (AX1 and AX2).
  • the same chemical substance (X, such as methylene blue dye) that provides different activities (A1, A2, and A3, such as cytotoxicity, local effect, and local synergy) can easily be prejudiced as the same drug (such as both Treat it as A1X and ignore A2X and A3X).
  • the effective cytotoxic effect of methylene blue dyes is denoted as A1, and the characteristic composition required for the effect is denoted as X1; its effective local effect (hence the local effect that is preferably maximized) is denoted as A2
  • the characteristic composition required for this action is denoted as X2; its effective local synergy is denoted as A3, and the characteristic composition required for this action is denoted as X3.
  • X1 with A1 was found in cell test suspension reactors and non-solid cancers, but A1X1 could not produce effective effects in tumor tissue reactors; in tumor tissue reactors, X1 with A1 was found A2 must be composed of X2 (extra-conventional concentration, such as ⁇ 2%), but the drug efficiency of A2X2 still needs to be improved.
  • the present invention provides the X3 technical solution to produce A3: low concentration ( ⁇ 2%, ⁇ 1%, or ⁇ 1%), A3X3 then only provides the local effect that is preferably minimized rather than maximized, but can provide effective local synergy The role of methylene blue dye.
  • A3X3 can prepare the composition of the present invention that far exceeds the expected technical effects (administration efficiency) of A1X1 and A2X2.
  • the characteristic activities of the methylene blue dye shared substance (Y) (such as the nutritional effect of nutrients, the immunomodulatory effect of immunomodulators, the cytotoxic effect of conventional antitumor drugs) are denoted as B1; nutrients and immunity
  • B1 The conventional anti-tumor activity (cytotoxic effect or other anti-tumor effect that can be exhibited by systemic administration, if any) other than the characteristic activity of the modulator is denoted as B2; its local synergy with the aforementioned methylene blue A3X3 is Denoted as B3, the latter is also known as the local synergist of A3X3 (denoted as B3Y).
  • the functional combination or composition of the composition of the present invention is not 1): the combination of methylene blue dye cytotoxicity (A1) and common compound characteristic activity (B1) (A1/B1) , Or A1X1/B1Y composition, nor 2): The combination (A1/B2) of methylene blue dye cytotoxicity (A1) and common anti-tumor activity (B2), or A1X1/B2Y composition, also Not 3): Combination of methylene blue dye local effect (A2) and common substance characteristic activity (B1) or conventional anti-tumor activity (B2) (A2/B1 or A2/B2), or A2X2/B1Y composition or A2X2 /B2Y composition, but 4): The combination (A3/B3) of the local synergistic effect of methylene blue dye (A3) and the local synergistic effect (B3) of the methylene blue dye, or the A3X3/B3Y composition.
  • this example and the following examples also confirm that the combination (A3/B3) or
  • the functional combination in the methylene blue dye/nutrient composition of the present invention is not the cytotoxic effect of methylene blue dye and the nutritional effect of nutrients (whether positive or side effects) or/and conventional anti-tumor effects (if If it exists) combination (A1/B1 or/and A1/B2), the local effect of non-methylene blue dye and the nutritional effect of nutrients (whether it is a positive or side effect) or/and conventional anti-tumor effect (if it exists) ) Combination (A2/B1 or/and A2/B2), but the local synergy of the methylene blue dyes that can provide nutrients (as chemical substances) and the nutrients that the nutrients can provide to the methylene blue dyes Combination (A3/B3).
  • the functional combination in the methylene blue dye/immunomodulator composition of the present invention is not the cytotoxic effect of the methylene blue dye and the immunomodulatory effect of the immunomodulator (whether it is a positive effect or a side effect) or/and
  • the methylene blue dye/conventional antitumor drug composition of the present invention is not a combination of the cytotoxic effect of methylene blue dye and the cytotoxic effect of conventional antitumor drugs (A1/B1), and it is also non-methylene blue.
  • Methylene blue dyes can provide conventional antitumor drugs (as Chemical substances) and the local synergistic combination (A3/B3) that the conventional anti-tumor drugs can provide to the methylene blue dye.
  • This example further studies the pharmacology of the local synergism of the composition of the present invention by optimizing the local synergy of the methylene blue dye to minimize rather than maximize the local action.
  • Experiment 3A The test animal was a nude mouse, the modeling cell was human pancreatic cancer cell (PANC-1), and the transplanted tumor was modeled at 1 ⁇ 10 6 cells/only under the skin of the animal's right axillary.
  • Successfully modeled test animals (nude mice bearing pancreatic cancer, with an average tumor volume of 218 mm 3 ) were randomly divided into 1 negative control group and 9 study groups.
  • the negative control is physiological saline, and the 9 study drugs are shown in the table below.
  • the medicines are all aqueous solutions, which are prepared according to the preparation method of Example 1.
  • the drugs are injected intratumorally.
  • the two drugs containing 50% acetic acid are administered once, and the injection volume is 100ul/only.
  • groups 3 and 8 a strong stimulus response to ethanol was observed in each animal.
  • groups 4 and 9 each animal observed the most powerful damaging response of acetic acid to the non-tumor tissue at the administration site, although its tumor inhibition rate was lower than that of groups 6 and 7, indicating that the latter had higher For the specificity of tumor tissue and better curative effect.
  • Test 3B The test animal was a nude mouse, the modeling cell was a liver cancer cell (MDA-MB231), and the transplanted tumor was modeled at 1 ⁇ 10 6 cells/only under the skin of the animal's right axillary.
  • the experimental animals (nude mice bearing liver cancer, with an average tumor volume of 127mm 3 ) that were successfully modeled were randomly divided into a negative control group and 14 study groups.
  • the negative control is physiological saline, and the 13 study drugs are shown in the table below.
  • the medicines are all aqueous solutions, which are prepared according to the preparation method of Example 1.
  • the drugs are injected intratumorally. The drug is administered once every 3 days, a total of 3 times, and the injection volume is 100 ⁇ l/mouse.
  • Ten days after the end of the medication the animals were euthanized, and the tumor weight (W) was measured after dissection, and the tumor inhibition rate (R) was calculated from the negative control group. The results are shown in the table below.
  • low-potency anti-tumor drug refers to drugs that are ineffective in conventional administration and are very effective in local administration, including non-steroidal anti-inflammatory compounds, quinoline compounds, artemisinin derivatives, and phenolic compounds.
  • quinine dihydrochloride is used as a representative of low-efficiency anti-tumor drugs
  • ferric chloride is used as a representative of metal compounds
  • 5-fluorouracil is used as a representative of effective anti-tumor drugs.
  • these drugs can be used as methylene blue dyes to maximize the local effect of further synergy.
  • the tumor inhibition rates of study groups 6, 8, and 10 are all above 115% of study group 4, which is in line with the synergy expectation of the prior art.
  • the tumor inhibition rates of study groups 6, 8, and 10 did not exceed the expected range of the theoretical simple additive effect (the actual/expected ratio q was both less than 1.15), indicating that these drugs ( The common effects of Y) on high-concentration methylene blue (X2) are all additive.
  • the tumor inhibition rates of study groups 7 and 9 are both above 115% of study group 5, and their tumor inhibition rates did not exceed their respective component single-drug groups (groups 2 and 5, respectively, As well as the expected range of the purely additive effect of groups 3 and 5) (the actual/expected ratio q is less than 1.15), it seems to further strengthen the above-mentioned mainstream view: drug Y is compared with low concentration methylene blue (X2) and high concentration sub The sharing of methyl blue (X2) only has a dynamic difference but no sharing mechanism difference, that is, all of the above-mentioned additive effects. Therefore, only by overcoming this mainstream view as a prejudice can we obtain a superadditive composition.
  • the two different methylene blue dyes studied are the prior art methylene blue dye (structure-activity B2X2) and the present invention methylene blue dye (structure-activity B3X3).
  • the prior art methylene blue dyes (B2X2) must be high concentrations (X2, preferably 2-6%) that can provide a high concentration (X2, preferably 2-6%) that preferably maximizes rather than minimizes the local effect (B2) dye.
  • high-concentration methylene blue dyes cannot, while low-concentration (X3, preferably 0.5%-1.5% or 0.5%-1.0%) methylene blue dyes may be
  • the local synergy (B3) is provided in the local sharing with the common object in the embodiment of the present invention, and the latter can only provide the local effect that is preferably minimized rather than maximized.
  • only methylene blue dyes whose local effects are preferably minimized rather than maximized can be used as "local active ingredients that can provide local synergistic effects" in the present invention.
  • the common substances studied include nutrients, immunomodulators, conventional anti-tumor drugs, chemical ablation agents, low-efficiency anti-tumor substances in the prior art, and metal compounds.
  • chemical ablation agents, conventional anti-tumor drugs, and low-efficiency anti-tumor substances all show strong local effects.
  • Nutrients and immunomodulators generally do not show strong local effects, and metal compounds have relatively small local effects.
  • whether these common compounds can provide local synergistic effects (rather than local effects) with methylene blue dyes does not seem to have much relationship with their own local effects. In other words, the local synergy of methylene blue dyes cannot be inferred based on the combination of local effects, and its intricacies are even required to overcome certain inherent concepts to be observed.
  • chemical ablation agents and low-efficiency anti-tumor substances with the strongest local effect can synergize high and low concentrations of methylene blue dyes (X2, X3) (for example, to increase the effect to more than 115%), but its actual effect
  • the /expected ratio q is all ⁇ 1.15
  • the synergistic effect is the additive effect of the local effects of the common components, rather than the local synergistic effect defined in the present invention. Therefore, chemical ablative agents and low-efficiency anti-tumor substances can be used as synergists for methylene blue dyes, but surprisingly they cannot be preferred as local synergists of methylene blue dyes in the present invention.
  • metal compounds with weaker local action is similar to chemical ablation agents and low-efficiency anti-tumor substances. They can also be used as synergists for methylene blue dyes, and they cannot be preferably used as methylene blue dyes in the present invention. Local synergist of dyes. In addition, the use of metal compounds with low-concentration methylene blue dyes even makes it difficult to produce the high efficacy required for the purpose of the present invention.
  • the local action between the chemical ablation agent and the metal compound, and even in many cases, the nutrient and immunomodulator, which is more likely to be the metal compound, and the low concentration methylene blue dye (X3) share the actual/expected ratio q up to Above 1.15, the shared effect is exactly the local synergistic effect not defined in the present invention. Therefore, the local synergists of methylene blue dyes in the present invention can be selected from nutrients and immunomodulators.
  • Example 4 Local synergistic composition (local synergistic amount ratio) is preferred
  • topical pharmaceutical composition a topical pharmaceutical composition
  • the topical pharmaceutical composition of the present invention must be completely different from the compositional characteristics of conventional pharmaceutical compositions: local synergistic dose ratio.
  • the term “synergistic dose ratio” refers to the combination of drugs X and Y necessary to produce a synergistic effect rather than an additive or antagonistic effect (actual/expected ratio q ⁇ 1.15).
  • the weight ratio of component X and component Y (W X :W Y ), the term “local synergistic amount ratio” means that it is not necessary in the conventional pharmaceutical composition (systemic administration), but in the local pharmaceutical composition (local Administering) the necessary synergistic amount ratio.
  • Test 4A The test animal was a mouse, the modeling cell was a sarcoma cell (S180), and the transplanted tumor was modeled at 1 ⁇ 10 6 cells/only under the skin of the animal's right axillary. Successfully modeled experimental animals (mice with sarcoma, with an average tumor volume of 112mm 3 ) were randomly divided into a negative control group and 40 study groups.
  • the negative control is physiological saline
  • the study drugs include: 4 kinds of methylene blue single drugs with varying concentrations (X%, X is the methylene blue concentration), 16 kinds of varying types and concentrations of other component single drugs (Y%, Y is Other component concentration), 20 compositions (X%/Y%) composed of varying concentrations of methylene blue (X%) and varying types and concentrations of other components (nutrients, conventional anti-tumor drugs) (Y%) ), their composition is shown in the table below.
  • the medicines are all aqueous solutions, which are prepared according to the preparation method of Example 1. All groups were injected intratumorally, once every 3 days for a total of 3 times.
  • the dose of each administration was: methylene blue ⁇ 100mg/kg, lysine ⁇ 1000mg/kg, DHA ⁇ 375mg/kg, glucose ⁇ 2250mg/kg , 5-Fluorouracil ⁇ 50mg/kg, injection volume ⁇ 150 ⁇ l.
  • the animals were euthanized on the 10th day after the treatment, the tumor weight was measured after anatomy, and the tumor inhibition rate was calculated from the negative control group.
  • the tumor inhibition rate of each study drug group is shown in the table below.
  • the data in the brackets is the average tumor inhibition rate of the X% methylene blue group, for example, the average tumor inhibition rate of the 0.15% methylene blue group is 3%
  • the data in the column bracket is the average tumor inhibition rate of Y% other component groups, for example, the average tumor inhibition rate of 1% lysine group is 5%
  • the data without brackets is the average tumor inhibition rate of X% methylene blue/Y% other component groups, for example, the average tumor inhibition rate of 3% methylene blue/1% lysine group is 45%
  • mice modeled with 1 ⁇ 10 6 breast tumor 4T1 cells per mouse mice modeled with 2 ⁇ 10 6 liver cancer H22 cells per mouse, and 1 ⁇ 10 6 liver cancer H22 cells per mouse.
  • mice modeled with 2 ⁇ 10 6 liver cancer H22 cells per mouse mice modeled with 2 ⁇ 10 6 liver cancer H22 cells per mouse
  • mice modeled with 1 ⁇ 10 6 liver cancer H22 cells per mouse mice modeled with 1 ⁇ 10 6 liver cancer H22 cells per mouse.
  • 106 CT26 mouse colon cancer cells modeling to 2 ⁇ 10 6 mouse B16-f10 th melanoma cells modeling each a similar effect can be observed.
  • Test 4B The test animal was a nude mouse, the modeling cell was a human liver cancer cell (MDA-MB231), and the transplanted tumor was modeled at 1 ⁇ 10 6 cells/only under the skin of the animal's right axillary.
  • Successfully modeled experimental animals (nude mice bearing liver cancer, with an average tumor volume of 163 mm 3 ) were randomly divided into a negative control group and 12 study groups.
  • the negative control is physiological saline.
  • the composition of the study drug is shown in the following table, including: 4 kinds of methylene blue single agents (X%) with varying concentrations, 4 kinds of immunomodulators (inactivated Saccharomyces cerevisiae) single agents ( Y%), and a combination of 4 different concentrations of immunomodulators and 4 different concentrations of methylene blue (X%/Y%).
  • the medicines are all water-containing liquids, which are prepared according to the preparation method of Example 1. Each group was treated twice, and the second treatment was 7 days after the first treatment, with an injection volume of 150 ⁇ l/mouse. The animals were euthanized 7 days after the end of the second medication. After dissection, the tumor tissue was stripped to determine the tumor weight, and the tumor inhibition rate was calculated from the negative control group.
  • the tumor inhibition rate of each study drug group is shown in the table below.
  • the data in the horizontal brackets is the average tumor inhibition rate of the X% methylene blue group, for example, the average tumor inhibition rate of the 0.15% methylene blue group is 10.1%
  • the data without brackets is the average tumor inhibition rate of the X% methylene blue/Y% immunomodulator composition group, for example, the average tumor inhibition rate of the 3% methylene blue/0.2% immunomodulator group containing group The rate is 75.2%.
  • the actual/expected ratio q of the composition did not increase with the increase of the methylene blue dye concentration. Instead, the local synergy ratio q was mainly dependent on the methylene blue dye concentration. The increase and decrease indicate that the local synergistic effect provided by methylene blue dyes is different or even deviated from its local effect. The actual/expected ratio q of the composition did not increase with the increase in the concentration of the shared materials, indicating that the local synergy provided by these shared materials is different from the local effect. Strength is the condition.
  • the local synergy ratio of methylene blue dye and its local synergist is actually what is needed for its local synergy Pharmacological dose ratio.
  • the local synergy ratio is >0.1/3 and ⁇ 40/0.15, preferably >0.1/1.8 and ⁇ 40/0.15, or >0.2/1 and ⁇ 40/0.30. in particular:
  • the local synergistic amount ratio of the nutrient/methylene blue dye composition (W nutrient /W methylene blue dye ) is 1/3 ⁇ W nutrient /W methylene blue dye ⁇ 40/0.15, preferably 1/3 ⁇ W nutrient /W methylene blue dye ⁇ 40/0.30, or 1/1.8 ⁇ W nutrient /W methylene blue dye ⁇ 40/0.30.
  • the local synergistic ratio of methylene blue dye / nutrient composition of amino acids is 1/3 ⁇ W-amino acid nutrient / W type dye methylene blue Dye ⁇ 20/0.15, preferably 1/3 ⁇ W amino acid nutrient /W methylene blue dye ⁇ 20/0.30, or 2/1.8 ⁇ W amino acid nutrient /W methylene blue dye ⁇ 20/0.30; lipid
  • the local synergistic quantity ratio of nutrient-like nutrients/methylene blue dye composition (W lipid nutrient /W methylene blue dye ) is 2/3 ⁇ W lipid nutrient /W methylene blue dye ⁇ 8/0.15, preferably 2/3 ⁇ W lipid nutrient /W methylene blue dye ⁇ 8/0.30, or 3/1.8 ⁇ W lipid nutrient /W methylene blue dye ⁇ 8/0.30; carbohydrate nutrient/methylene blue dye
  • Synergistic ratio topical immunomodulators / methylene blue dye composition was 0.2 / 3 ⁇ W immunomodulators / W methylene blue dye ⁇ 20 / 0.15 , preferably from 0.5 / 3 ⁇ W immunomodulators / W methylene blue dye ⁇ 20 / 0.50, or 0.5 / 1 ⁇ W immunomodulatory agents / W methylene blue dye ⁇ 20 / 0.50.
  • the local synergistic amount ratio of the conventional antitumor drug/methylene blue dye composition (W conventional antitumor drug /W methylene blue dye ) is 0.1/3 ⁇ W conventional antitumor drug /W methylene blue dye ⁇ 1.5 /0.15, preferably 0.3/3 ⁇ W conventional anti-tumor drug /W methylene blue dye ⁇ 1.5/0.15, or 0.3/1.8 ⁇ W conventional anti-tumor drug /W methylene blue dye ⁇ 1.5/0.30.
  • Example 5 The effective local synergistic concentration is optimized
  • the dosage concentration can be adjusted by the clinician (for example, 1-5 times difference), which is not a feature of the pharmaceutical composition, at least not an important feature that can lead to changes in the shared mechanism of action.
  • the topical pharmaceutical composition of the present invention has a composition characteristic completely different from conventional pharmaceutical compositions: local effective synergistic concentration.
  • the term “synergistic concentration” refers to the group in the administration composition necessary for the drugs X and Y to have a synergistic effect instead of an additive or antagonistic effect (actual/expected ratio q ⁇ 1.15).
  • concentration of component X or component Y refers to the sub-necessary synergistic concentration in conventional pharmaceutical compositions (systemic administration) but necessary in local pharmaceutical compositions (local administration).
  • Local effective synergistic concentration refers to the local synergistic concentration required for the composition X/Y to show more competitive efficacy compared with existing drugs.
  • Test 5A The test animal was a BALB/c mouse, the modeling cells were breast cancer 4T1 cells, and the transplanted tumor was modeled at 1 ⁇ 10 6 cells/only under the skin of the animal's right axillary.
  • Successfully modeled experimental animals (mice bearing breast cancer, with an average tumor volume of 119mm 3 ) were randomly divided into 1 negative control group and 9 study groups.
  • the negative control is physiological saline, and the 9 study drugs are shown in the table below.
  • the medicines are all aqueous solutions, which are prepared according to the preparation method of Example 1.
  • the drugs are injected intratumorally. The drug is administered once every 3 days, a total of 3 times, and the injection volume is 100 ⁇ l/mouse.
  • the animals were euthanized 10 days after the end of the medication, and the tumor weight (W) was measured after anatomy, and the tumor inhibition rate (R) was calculated from the negative control group of each medication mode. The results are shown in the following table.
  • the dose ratio of the methylene blue dye/topical synergistic composition pharmaceutical composition used in the composition groups 4 and 9 is the same but the dose concentration is different. They not only show different therapeutic effects, but also show a common effect. Different mechanisms.
  • the actual/expected ratio q of the former is >1.15, showing local synergy; the actual/expected ratio q of the latter is ⁇ 1.15 and >0.85, showing an additive effect.
  • the composition groups 5 and 10 also showed different mechanisms of shared action. It is especially important that the efficacy of the composition obviously depends on whether they show a local synergistic effect.
  • the local synergistic concentration of methylene blue dye is> 0.25%, and its effective local synergistic concentration is preferably ⁇ 0.5%.
  • Test 5B The test animal was a BALB/c mouse, the modeling cells were breast cancer 4T1 cells, and the transplanted tumor was modeled at 1 ⁇ 10 6 cells/only under the skin of the animal's right axillary.
  • Successfully modeled experimental animals (mice bearing breast cancer, with an average tumor volume of 386mm 3 ) were randomly divided into 1 negative control group (01) and 6 drug research groups (1-6).
  • the negative control is physiological saline, and the study drugs are shown in the table below.
  • human immunoglobulin is an immunoglobulin isolated from human plasma for laboratory use, and the main component is IgG.
  • the medicines are all aqueous solutions, which are prepared according to the preparation method of Example 1.
  • Each test group received intratumoral injection, once every 3 days for a total of 3 times.
  • the amount of injection is shown in the table below.
  • the efficacy depends on the dose rather than the concentration of the drug.
  • the clinically safe blood drug concentration is usually very low (for example, less than 10ug/ml), which is much lower than the usual preparation concentration.
  • the dose ratio of the methylene blue dye/immunomodulator composition used in the composition groups 5 and 6 is the same, the dose is the same, but the dose concentration is different. They not only show different therapeutic effects, but also share Different mechanisms of action.
  • the actual/expected ratio q of the former is> 1.15, showing local synergy; the latter does not show meaningful efficacy, let alone local synergy.
  • the tumor inhibition rate of the composition group 5 is 663% of that of the composition group 6, which also shows that the corresponding compositions have completely different common pharmacology.
  • the local synergistic concentration of methylene blue dye is> 0.30%, and its effective local synergistic concentration is preferably ⁇ 0.9%.
  • Test 5C The test animal was a BALB/c mouse, the modeling cell was a sarcoma cell S180 cell, and the transplanted tumor was modeled at 1 ⁇ 10 6 cells/only under the skin of the animal's right axillary.
  • Successfully modeled experimental animals (mice bearing sarcoma, with an average tumor volume of 132mm 3 ) were randomly divided into a negative control group, a positive control group and 7 study groups.
  • the negative control is physiological saline
  • the positive control is 1% 5-fluorouracil
  • the study drug contains 20% lysine and methylene blue (20% lysine/X%) with varying concentrations (X%) as shown in the table below. Methylene blue).
  • the medicines are all aqueous solutions, which are prepared according to the preparation method of Example 1.
  • Each group was injected intratumorally, once every 3 days for a total of 3 times.
  • the dose of each administration was: methylene blue ⁇ 75 mg/kg, 5-fluorouracil 50 mg/kg, and injection volume ⁇ 150 ⁇ l.
  • the tumor weight (W) was determined after the animal was dissected 10 days after the end of the medication, and the tumor inhibition rate (R) was calculated from the negative control group. The results are shown in the table below.
  • the methylene blue dye in the methylene blue dye/amino acid nutrient composition also shows a characteristic when it meets its local synergistic amount ratio and local synergistic concentration: it makes the drug significantly exceed the conventional antitumor drugs Group (positive control group) the concentration threshold of the drug effect, that is, the effective local synergistic concentration threshold.
  • the concentration threshold of the drug effect that is, the effective local synergistic concentration threshold.
  • the effective local synergistic concentration of methylene blue dye is 0.35%-2%, preferably 0.35%-1.5% or 0.50%-1.5% (w/v).
  • Test 5D The successfully modeled test animals (mice bearing S180 cells, with an average tumor volume of 128mm 3 ) were randomly divided into a negative control group (0), a positive control group (7) and 6 study groups (1-6).
  • the negative control is normal saline, and the positive control is 1% 5-fluorouracil.
  • the composition of the study drug is shown in the table below, which contains a local synergist (arginine) of varying concentration (X%, w/v) and a fixed concentration (1%) methylene blue (X% arginine/1% methylene blue).
  • the medicines are all aqueous solutions, which are prepared according to the preparation method of Example 1. Each group was injected intratumorally, once every 3 days for a total of 3 times.
  • the dose of each administration was: arginine ⁇ 1500 mg/kg, 5-fluorouracil 50 mg/kg, and injection volume ⁇ 150 ⁇ l.
  • the animals were euthanized 5 days after the end of the medication, and the tumor weight (W) was measured after anatomy, and the tumor inhibition rate (R) was calculated from the negative control group. The results are shown in the following table.
  • the amino acid nutrients in the methylene blue dye/amino acid nutrient composition when meeting its local synergistic amount ratio and local synergistic concentration, also show a feature: making the drug significantly exceed the conventional anti-tumor drug group ( Positive control group) the concentration threshold of the drug effect, that is, the effective local synergistic concentration threshold.
  • the concentration threshold of the drug effect that is, the effective local synergistic concentration threshold.
  • the effective local synergistic concentration of amino acid nutrients is ⁇ 5%, preferably 5%-35% or 15%-35% (w/v).
  • a necessary technical condition for the application of the combination of methylene blue dye and its local synergist as a local synergistic active ingredient is: the preparation and composition of the composition must meet the methylene blue dye
  • the local synergistic concentration of its local synergist preferably the effective local synergistic concentration, specifically:
  • the local administration concentration (w/v) of the methylene blue dye is less than or equal to 2%, preferably 0.3%-1.8%, 0.5%-1.0%, or 0.5%-0.9%;
  • the local administration concentration (w/v) of the conventional antitumor drug is greater than 20% of its saturated concentration, preferably 30%-100% of its saturated concentration;
  • the local administration concentration (w/v) of the immunomodulator is greater than 0.25%, preferably 0.5%-30%, wherein when the immunomodulator includes immunoglobulin, the administration concentration (w/v) of the immunoglobulin /v) ⁇ 0.25%, preferably 0.25-30% or 1-30%; when the immunomodulator includes a probiotic component, the administration concentration (w/v) of the probiotic component is ⁇ 0.35%, preferably When the immunomodulator includes an immunomodulatory peptide, the administration concentration (w/v) of the immunomodulatory peptide is ⁇ 3%, preferably 3-30%;
  • the local administration concentration (w/v) of the nutrient is greater than 3%, preferably 3%-45%, wherein when the nutrient includes a carbohydrate nutrient, the administered concentration of the carbohydrate nutrient (w/v) ⁇ 20%, preferably 20-40%; when the nutrient includes lipid nutrient, the administration concentration (w/v) of the lipid nutrient is ⁇ 4%, preferably 4-25%; when the nutrient includes amino acid nutrient
  • the administration concentration (w/v) of the amino acid nutrients is ⁇ 3%, preferably 3-25%, more preferably 5-25% or 15-35%.
  • Low-concentration methylene blue dyes can be used as detoxification active ingredients or conventional anti-tumor ingredients (A1X1) to prepare conventional pharmaceutical compositions, and high-concentration (or local effects maximize rather than minimize) methylene blue dyes can provide effective Locally acting active ingredients (A2X2) should be used in the preparation of locally acting pharmaceutical compositions, while low concentrations (or local effects are minimized rather than maximized) methylene blue dyes alone are not acceptable, only in combination with their local synergists It can be used as a local active ingredient (A3X3) that provides effective local synergistic effect and is applied to prepare a local synergistic pharmaceutical composition. Using the same dose of methylene blue dye, the local synergistic drug composition shows the tumor tissue destruction efficacy, so that the scope of indications exceeds the expectations of the local drug composition and the conventional drug composition.
  • Methylene blue dyes can only provide the above-mentioned local synergistic effects when they are shared with the preferred common substance (its local synergist).
  • the shared effect of methylene blue dyes and their local synergists must be local synergy, not an additive effect of local effects, so it is impossible to find clues taught in the prior art (for example, shared enhancement of local effects) that can provide the local Synergistic commons.
  • the local synergist is preferably selected One or more from the following groups: nutrients, immunomodulators, conventional anti-tumor drugs.
  • compositions of the combination are in addition to the above-mentioned component characteristics (X is selected from the local effect).
  • Minimized but not maximized methylene blue dye, Y is selected from nutrients, immunomodulators, or/and conventional anti-tumor drugs), and also includes the synergistic ratio between X and Y.
  • the compositional characteristics of the locally synergistic pharmaceutical composition may also include an effective synergistic concentration different from its conventional pharmaceutical composition or a locally acting pharmaceutical composition, for example, a methylene blue dye that is different from the effective synergistic concentration of the locally acting pharmaceutical composition. concentration.
  • composition of the present invention show higher specificity for tumor tissues.
  • composition of the present invention is compared with a conventional chemical ablation agent (50% acetic acid).
  • the combination of the methylene blue and the topical synergist which can be used as a local synergistic active component, has strong coupling properties, and it is difficult to derive from common sense.
  • the following experiments further study the coupling, in order to facilitate the optimization of the essential components of the composition of the present invention.
  • Test 6A The test animal was a BALB/c nude mouse, the modeling cells were melanoma cells, and the transplanted tumor was modeled at 5 ⁇ 10 6 cells/only under the skin of the animal's right axillary.
  • Successfully modeled test animals (nude mice bearing melanoma, with an average tumor volume of 238mm 3 ) were randomly divided into 1 negative control group (0) and 9 drug study groups (1-9).
  • the negative control is physiological saline, and the study drugs are shown in the table below.
  • the medicines are all aqueous solutions, which are prepared according to the preparation method of Example 1. Each group was given the drug once every 3 days for a total of 3 times, intratumoral injection was performed, and the injection volume was 125 ⁇ l/mouse. Three days after the end of the medication, the animals were euthanized, and the tumor weight (W) was measured after dissection, and the tumor inhibition rate (R) was calculated from the negative control group. The results are shown in the following table.
  • the tumor weights of the composition groups 6-8 are statistically significant compared with the negative control group (all p ⁇ 0.05), and their respective actual/expected ratios q are all> 1.15, all showing Obvious local synergy.
  • the tumor weight of the composition group 9 was not statistically significant (p>0.05), and it did not show significant pharmacological effects (tumor inhibition rate less than 20%), so it did not show obvious local synergy .
  • Test 6B The test animal was a BALB/c mouse, the modeling cell was a sarcoma S180 cell, and the transplanted tumor was modeled at 1 ⁇ 10 6 cells/only under the skin of the animal's right axillary.
  • Successfully modeled test animals (S180 cell-bearing mice with an average tumor volume of 151mm 3 ) were randomly divided into negative control group, positive control group and 15 study groups.
  • the negative control was normal saline
  • the positive control was 1% 5-fluorouracil
  • the study drug was 1% methylene blue/10% amino acid nutrient composition (1% methylene blue/Y%).
  • the amino acid nutrients in the compositions used in study groups 1-14 were: arginine, glycine, cysteine hydrochloride, valine, threonine, proline, histidine hydrochloride, benzene Alanine, lysine, leucine, alanine, glutathione, serine, alanine (aminoacyl)-glutaminamide dipeptide, the study group 15 medication is 1% methylene blue/5% Arginine/5% Glycine.
  • the medicines are all aqueous solutions, which are prepared according to the preparation method of Example 1. Each group was injected intratumorally, once every 3 days for a total of 3 times, and the volume of each injection was ⁇ 150 ⁇ l.
  • the animals were euthanized 10 days after the end of the medication, and the tumor weight was measured after anatomy, and the tumor inhibition rate (R) was calculated from the negative control group.
  • the tumor inhibition rate of the positive control group was 51%, and the results of the study group are shown in the table below.
  • the amino acid nutrient is preferably selected from the following amino acids and one of the following amino acids Or multiple amino acid derivatives: arginine, glycine, cysteine, threonine, proline, lysine, leucine, alanine, serine, glutamic acid, more preferably selected from The following amino acids and amino acid derivatives containing one or more of the following amino acids: arginine, glycine, cysteine, lysine, alanine, serine, and glutamic acid.
  • Test 6C The test animal was a BALB/c nude mouse, the modeling cells were human pancreatic cancer PANC-1 cells, and the transplanted tumor was modeled at 1 ⁇ 10 6 cells/only under the skin of the animal's right axillary.
  • Successfully modeled test animals (nude mice bearing pancreatic cancer, with an average tumor volume of 207mm 3 ) were randomly divided into negative control group (group 0), methylene blue single-agent group (group 01), and 8 local synergistic single-agents Groups (groups 1-8) and 8 methylene blue/topical synergistic composition groups (groups 9-16).
  • the negative control is physiological saline
  • the local synergists are as shown in the following table, including: single local synergist (groups 1, 2, 9, 10), acidified nutrients (groups 3, 4, 11, 12), neutral pH Chemical amino acid nutrients (groups 5, 13), nutrients/conventional anti-tumor drug complexes (groups 6, 14), nutrients/immunosuppressive compound (groups 7, 15), immunosuppressant/conventional anti-tumor drug complexes (Group 8, 16).
  • the medicines are all water-containing liquids, which are prepared according to the preparation method of Example 1. Each group was injected intratumorally once, and the injection volume was 170 ⁇ l.
  • the animals were euthanized 7 days after the treatment, and the tumor weight (W) was measured after anatomy, and the tumor inhibition rate (R) was calculated from the negative control group, and the actual/expected ratio (q) was calculated from the tumor inhibition rate.
  • W tumor weight
  • R tumor inhibition rate
  • q actual/expected ratio
  • the tumor weights of 9-16 of each composition group are statistically significant compared with the negative control group (all p ⁇ 0.05), and their actual/expected ratio q are all >1.15, all showing local synergy effect. More specifically, the composition groups 11-16 showed higher efficacy than the composition groups 9 and 10.
  • the mainstream view of anti-tumor drug composition in the prior art is: the probability of synergistic effect between X and a drug Y is very low, while the probability of synergistic effect with drug compound Y/Z is lower, and the compound is on the contrary Increase the risk of drug antagonism. As a result, there is almost no anti-tumor three-component synergistic composition clinically.
  • a preferred technical solution of the topical synergist in the composition of the present invention is: a multi-component complex containing the topical synergist (abbreviated in the present invention) Is a local synergist complex), more preferably the following local synergist complex: homogeneous local synergist complex, different local synergist complex, a single local synergist or a pH adjusting complex of a local synergist complex, wherein :
  • the local synergistic complex of the same type includes multiple local synergists of the same type, including nutrient complexes, immunosuppressant complexes, and conventional anti-tumor drug complexes, wherein the nutrient complexes include complexes of multiple different nutrients Compound (for example, amino acid nutrient/carbohydrate nutrient complex) and a complex of multiple same nutrients (for example, amino acid nutrient complex, carbohydrate nutrient complex), wherein the amino acid nutrient complex includes a compound with acid-basic amino acid nutrient (Eg arginine/acid-resistant complex, glutamate/glycine complex, etc.) and iso-acid-basic amino acid nutrient complex, wherein the iso-acid-basic amino acid nutrient complex is preferably such that The pH of the aqueous liquid tends to be neutralized iso-acid-basic amino acid nutrient complex (e.g.
  • the term "same acid-base complex” refers to a complex of multiple local synergists that are both acidic or the same in a 1% aqueous solution; the term “iso acid-base complex” "Refers to a complex of local synergists of acidity and alkalinity respectively in 1% aqueous solution;
  • the different types of local synergistic complexes include multiple different types of local synergists, including, for example, nutrient/conventional anti-tumor drug complexes (such as arginine/lysine/conventional anti-tumor drugs, arginine/conventional Anti-tumor drugs, etc.), nutrient/immunosuppressant complexes (e.g. arginine/immunoglobulin, arginine/probiotic components, arginine/lysine/RNA, etc.), immunosuppressants/ Conventional anti-tumor drug complexes (such as probiotic components/conventional anti-tumor drugs), etc.
  • nutrient/conventional anti-tumor drug complexes such as arginine/lysine/conventional anti-tumor drugs, arginine/conventional Anti-tumor drugs, etc.
  • nutrient/immunosuppressant complexes e.g
  • the pH-adjusting complex of the local synergist includes the acidification (local synergist/acid) or alkalinization (local synergist/base) complex of the local synergist, preferably so that the local synergist is aqueous liquid
  • the pH change value of is> 1, more preferably a complex (such as arginine/acetic acid complex) that makes the pH of the aqueous liquid of the local synergistic neutralize, wherein the acid in the acidification is preferably an organic acid ( For example, acetic acid), the alkali in the alkalization is preferably a pharmaceutically acceptable pH adjusting alkali (for example, sodium hydroxide, sodium bicarbonate, etc.).
  • the successfully modeled nude mice bearing human cancer cells were randomly divided into a negative control group and 5 study groups (groups A, B, C, D, and E).
  • the corresponding negative control is physiological saline
  • the 5 study drugs are: A (1% methylene blue/5% DHA/5% acetic acid), B (1% methylene blue/20% arginine), C (1% methylene blue/10% glycine/10% lysine), D (1% methylene blue/30% glucose), E (1% methylene blue/1% 5-fluorouracil).
  • the medicines are all aqueous solutions, which are prepared according to the preparation method of Example 1.
  • Each group was injected intratumorally, once every 3 days, a total of 3 times, each time 100-150 ⁇ l/mouse.
  • the animals were euthanized, the tumor weight was measured after anatomy, and the tumor inhibition rate was calculated from the respective negative control group.
  • the successfully modeled nude mice bearing human head and neck cancer cells (F ⁇ da) (the average tumor volume is 169mm 3 ) were randomly divided into a negative control group and 5 study groups (groups A, B, C, D, and E) ).
  • the tumor inhibition rates of groups A, B, C, D, and E were 76%, 82%, 74%, 88%, 70%, respectively, which all met the generally considered effective anti-tumor standards (tumor inhibition rate ⁇ 40%).
  • CNE1 human nasopharyngeal carcinoma cells
  • compositions of the present invention prepared in Example 1 for example, the composition in Table 2
  • similar results can be obtained in the application of the above-mentioned tumor treatments.
  • Jinghuai treatment requires precise determination of the relationship between anti-tumor drugs and their indications.
  • the safety dose of methylene blue dye (for example, 1 mg/kg body weight or 80 mg/person in the technical solution for maximum safety) is the decisive factor for the safety dose of the composition containing it. factor.
  • the administration volume (for example, 8 ml of a drug containing 1% (w/v) methylene blue, or 4 ml of a drug containing 2% (w/v) methylene blue) is directly related to the drug effect, which is
  • the technical solution for maximizing the effectiveness is closely related to the characteristics of the tissue within the tumor and the size of the tumor body, for example, it is preferably equal to or greater than the volume of the tumor body to be administered. Therefore, the composition of the present invention is greatly superior to the medicinal effect (also referred to as methylene blue dye drug efficiency in the present invention) provided by a unit methylene blue dye dose (e.g. mg) due to its local synergistic effect
  • methylene blue-containing dye composition of the prior art enables the composition of the present invention to expand new indications.
  • heterogeneity is a core feature of tumors
  • the indications defined by this heterogeneity are then a core feature of tumor therapy drugs. It is generally believed that the heterogeneity of cancer cells is the basis for the indications of anticancer drugs.
  • the following experiments further illustrate the characteristics of the composition of the invention and the indication characteristics corresponding to the characteristics of the composition under completely different conditions of tumor heterogeneity through a comparative study of the composition of the invention and the composition of the prior art.
  • Test 8A The different sharing mechanisms of different topical compositions against diseased tissues of different compositions were studied.
  • the test animals are BALB/c nude mice.
  • the rat benign fibroma tissue fragments (approximately 20 mm 3 /piece) of approximately 300 mm 3 / only under the skin of the animal's right axillary tissue transplantation tumor modeling for series I use.
  • human breast cancer cells (MDA-MB231) were used at 1 ⁇ 10 6 cells/only under the skin of the right axilla of other animals in the same batch to model cell transplantation tumors for use in Series II.
  • the main component of rat breast benign fibroma tissue is fibrous connective tissue.
  • the tumor weights of each series of composition groups 5-7 are statistically significant compared with the negative control group (all p ⁇ 0.05).
  • the actual/expected ratio q of each series of composition groups 5 and 6 are both> 1.15
  • the actual/expected ratio q of each series of composition group 7 are both ⁇ 1.15, indicating that the former pharmaceutical compositions all show local synergy
  • the latter's pharmaceutical composition did not show local synergistic effects.
  • the composition of the present invention can solve the problem that cannot be solved by the methylene blue-containing dye composition of the prior art at the same dose: the effective destruction of tumor tissues rich in non-cancer components.
  • a preferred solution for the indications of the composition of the present invention is the treatment of nodules related to malignant tumors.
  • malignant tumor-related nodules refer to any nodules that should be paid attention to clinically for the treatment of malignant tumors, such as those that have not been detected (for example, undetected due to undetected, misdetected, or missed detection) It is, but cannot be 100% excluded as malignant tumor nodules, or nodules that may develop into malignant tumor tumors.
  • Test 8B The test animals were BALB/c nude mice. According to known techniques, the easy-to-operate human pancreatic cancer tissue is broken into about 50 mm 3 /piece, and then the tissue transplantation tumor is modeled under the skin of the animal's right axillary at a rate of about 300 mm 3 /piece for use in Series II. After the transplantation was successful, human pancreatic cancer cells (PANC-1) were modeled under the skin of the right axillary of the same batch of other animals at 1 ⁇ 10 6 cells/only for use in series I.
  • PANC-1 human pancreatic cancer cells
  • the animals were euthanized 7 days after the treatment, the tumor weight was measured after anatomy, and the tumor inhibition rate (R) was calculated from the negative control group, and the actual/expected ratio (q) was calculated from the tumor inhibition rate.
  • the results of the study group are shown in the table below .
  • the average interstitial ratios of series I and II tumors were 11.3% and 19.7%, respectively.
  • the term "larger interstitial ratio" refers to an interstitial ratio of ⁇ 20%, and an interstitial ratio of ⁇ 20% is referred to as a smaller interstitial ratio.
  • tumor parenchyma such as cancer cells
  • interstitium such as connective tissue, fibrous structure, etc. intercellular substance in non-cancerous cells.
  • the quantity and composition of the stroma is one of the key factors for tumor occurrence and development.
  • Mesenchymal ratio can then characterize this tumor heterogeneity. More and more clinical data show that under roughly the same treatment conditions, the tumor-to-interstitial ratio is an independent factor of tumor treatment efficacy (or prognosis).
  • the heterogeneity based on the interstitial ratio can be so great that the same anti-tumor drug can have very different effects on the same tumor cells but with different interstitial ratios.
  • tumors with different mesenchymal ratios must be regarded as two different tumors at this time.
  • 5-fluorouracil is effective when applied to human pancreatic cancer cell transplantation tumors (usually relatively small in interstitial) in animal experiments (tumor inhibition rate ⁇ 40%), but it is ineffective when applied to clinical human pancreatic pancreatic cancer (usually relatively large in interstitial) (Tumor inhibition rate ⁇ 20%). Therefore, animal human pancreatic cancer cell transplantation tumor is, but clinical human pancreatic pancreatic cancer is not an indication for 5-fluorouracil.
  • the tumor weights of each series of composition groups 5-7 are statistically significant compared with the negative control group (all p ⁇ 0.05).
  • the actual/expected ratio q of each series of composition groups 5 and 6 are both> 1.15
  • the actual/expected ratio q of composition group 7 are both ⁇ 1.15, indicating that the pharmaceutical compositions of the former all show local synergistic effects.
  • the latter pharmaceutical composition did not show local synergy.
  • the methylene blue-containing dye composition of the present invention can solve the problem that cannot be solved by the methylene blue-containing dye composition of the prior art at the same dose: tumor tissue with a larger interstitial ratio Effective destruction.
  • a preferred solution for the indications of the composition of the present invention is the treatment of tumors with larger stromal ratios and tumors containing the tumors.
  • Test 8C The test animals were BALB/c nude mice.
  • the human liver cancer cell MDA-MB231 was modeled under the skin of the animal's right axillary at 1 ⁇ 10 6 cells/only according to known techniques for use in Series II. After the transplantation is successful (the tumor is visible), the hepatoma cell HepG2 is then used at 1 ⁇ 10 6 cells/only under the skin of the right axillary of the same batch of other animals to model the cell transplantation tumor for series I.
  • the tumor weight of each series composition group 9 is not statistically significant compared with the negative control group (all p>0.05), and the tumor weights of each series composition group 8, 10-12 are compared with the negative control group The comparisons are statistically significant (all p ⁇ 0.05).
  • the actual/expected ratio q of each series of composition groups 8 and 11 are both> 1.15, while the actual/expected ratio q of composition groups 10 and 12 is ⁇ 1.15, indicating that the pharmaceutical composition of the former shows that the pharmaceutical composition of the former and the latter The pharmaceutical composition did not show local synergy.
  • the same partial synergistic composition group (groups 8 and 11) showed a higher actual/expected ratio q in series II than in series I.
  • the term "larger tumor” when applied to humans refers to tumors with an average size of ⁇ 2.0cm, while tumors with an average size of ⁇ 2.0cm are called smaller tumors. body.
  • the increase in tumor size is the result of a series of complex living environment evolution that is caused by the proliferation of tumor cells and the growth of non-tumor components in the tumor.
  • tumor cell metabolism requires redistribution of blood vessels, which in turn requires tumor cell proliferation to preferentially proceed on the surface where redistribution is most likely to occur.
  • the enlargement of the tumor also increases the influence of non-tumor components (for example, the inter-fibrous structure) on the growth of the tumor tissue.
  • the average tumor size can then characterize this tumor heterogeneity. More and more clinical data show that under roughly the same treatment conditions, the average tumor size is an independent factor of the therapeutic effect (or prognosis) of local tumor treatment.
  • the heterogeneity based on the average size of the tumor can be so great that the same local drug or physical method can have very different therapeutic effects on tumors of the same tumor cells but of different sizes.
  • tumors of different sizes must be regarded as two different tumors at this time.
  • the classical chemical ablation agent applied to the transplanted tumors of human liver cancer cells (usually small tumors) in animal trials is effective (tumor inhibition rate ⁇ 40%), but it is no longer regarded as clinically larger tumors of human liver cancer (average size). ⁇ 2.0cm) effective drugs. Therefore, the transplanted tumor of animal human liver cancer cells is, but the clinically larger human liver cancer is not an indication for the classic chemical ablation agent.
  • the methylene blue-containing dye composition of the present invention can solve the problem that cannot be solved by the methylene blue-containing dye composition of the prior art at the same dose: effective destruction of tissues in larger tumors.
  • a preferred solution for the indications of the composition of the present invention is the treatment of larger tumor bodies and tumors containing the tumor bodies.

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Abstract

L'invention concerne une combinaison contenant un colorant bleu de méthylène en tant qu'ingrédient actif local et un médicament synergique local ou un synergiste de celui-ci et une utilisation de celle-ci dans la préparation d'une composition de médicament local pour le traitement du cancer, tel qu'une tumeur solide maligne, et un procédé d'utilisation de ladite combinaison contenant un colorant bleu de méthylène en tant que principe actif local et un médicament synergique local ou un synergiste de celui-ci pour traiter le cancer, tel qu'une tumeur solide maligne.
PCT/CN2021/076749 2020-02-18 2021-02-18 Composition pharmaceutique contenant un colorant bleu de méthylène et son utilisation WO2021164706A1 (fr)

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EP21874572.7A EP4223313A1 (fr) 2020-09-30 2021-09-30 Composition pharmaceutique comprenant une combinaison de neutralisation acide-base et son application
US18/247,359 US20230414617A1 (en) 2020-09-30 2021-09-30 Pharmaceutical composition comprising acid-base neutralization combination and application thereof
JP2023519733A JP2023543858A (ja) 2020-09-30 2021-09-30 酸－塩基中和合剤を含む医薬組成物及びその使用
PCT/CN2021/122041 WO2022068918A1 (fr) 2020-09-30 2021-09-30 Composition pharmaceutique comprenant une combinaison de neutralisation acide-base et son application
PCT/CN2022/076823 WO2022174812A1 (fr) 2021-02-18 2022-02-18 Composition pharmaceutique comprenant de l'immunoglobuline et son utilisation

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PCT/CN2020/075765 WO2021163897A1 (fr) 2020-02-18 2020-02-18 Composition pharmaceutique contenant un colorant bleu de méthylène, un élément nutritif et/ou un médicament antitumoral classique, et son application
CN202011059746.1 2020-09-30
CN202011059699.0A CN114344333A (zh) 2020-09-30 2020-09-30 动物非致病性细胞相关组分的应用和包含该组分的药物组合物
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CN202011064448.1A CN114306392A (zh) 2020-09-30 2020-09-30 益生菌组分的应用以及包含益生菌组分的药物组合物
CN202011059746.1A CN114306612A (zh) 2020-09-30 2020-09-30 非致病性细胞相关组分的应用以及包含非致病性细胞相关组分的药物组合物

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WO2023050297A1 (fr) * 2021-09-30 2023-04-06 成都夸常奥普医疗科技有限公司 Composition pharmaceutique topique, application et kit

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