WO2022174812A1 - Pharmaceutical composition comprising immunoglobulin, and use thereof - Google Patents

Pharmaceutical composition comprising immunoglobulin, and use thereof Download PDF

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Publication number
WO2022174812A1
WO2022174812A1 PCT/CN2022/076823 CN2022076823W WO2022174812A1 WO 2022174812 A1 WO2022174812 A1 WO 2022174812A1 CN 2022076823 W CN2022076823 W CN 2022076823W WO 2022174812 A1 WO2022174812 A1 WO 2022174812A1
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Prior art keywords
acid
pharmaceutical composition
tumor
local
immunoglobulin
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PCT/CN2022/076823
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French (fr)
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
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Priority claimed from PCT/CN2021/076749 external-priority patent/WO2021164706A1/en
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 WO2022174812A1 publication Critical patent/WO2022174812A1/en

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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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/736—Glucomannans or galactomannans, e.g. locust bean gum, guar gum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • 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 present application discloses a novel application of immunoglobulin, a topical pharmaceutical composition comprising an immunoglobulin suitable for the application and a synergist thereof, and a method of using the pharmaceutical composition to treat local pathological diseases.
  • Solid tumors are often used as models for the study of localized disease, especially refractory localized disease, supported by a large amount of research work.
  • Solid tumor is a tumor disease with tumor symptoms, and the tumor body is a characteristic diseased tissue containing tumor cells. Taking pancreatic cancer tumor as an example, pancreatic cancer cells only account for about 30% of the volume. It can be seen that in addition to tumor cells, there are often a larger number of other components (sometimes also referred to as tumor cell microenvironment) in tumor tissue, including other various cells (such as fibroblasts), various intercellular substances (such as fiber), various pipes, etc.
  • Topical administration has the advantage of physically targeting the drug. Therefore, it was once believed that local administration of cytotoxic drugs can increase their intratumoral concentration, thereby improving their efficacy. However, topical administration of cytotoxic drugs did not show a disparate improvement in efficacy. Merely increasing its intratumoral concentration does not seem to greatly improve the efficiency of targeting cancer cells in intratumoral tissues, but significantly increases local irritation. Thus, cytotoxic drugs are still clinically administered almost systemically, except for recourse to their sustained-release forms. Clinically used topically active drugs such as chemical ablatives (high-purity ethanol, high-concentration acid-base, weak-acid-strong-base salt) are not characterized by cell destruction but tissue destruction.
  • chemical ablatives high-purity ethanol, high-concentration acid-base, weak-acid-strong-base salt
  • cytotoxic drugs Compared with cytotoxic drugs, they have almost no systemic efficacy targeting cancer cells, but often show higher local efficacy. However, they are generally strong disruptors that do not sufficiently distinguish the target tissue from other tissues. This makes them practically applicable to the intervention volume (for example, the amount of acid and alkali does not exceed 0.2ml/kg) and the intervention site is very limited (for example, the limitation of the organ where the tumor is located, the ablation of the tumor edge, etc.). In addition, they usually have no mid- to long-term effect and are easily rebound. As a result, chemical ablative agents have been clinically faded from malignant solid tumors in the past decade. In fact, there are almost no local drugs with high local safety and short-term local efficacy in clinical practice, and there is no local drug with medium and long-term efficacy.
  • the purpose of the present invention is to provide a higher comprehensive performance (for example, local safety, local short-term efficacy, or/and medium and long-term efficacy) for the prevention and treatment of local diseased diseases, especially refractory local diseased diseases. topical medication.
  • a higher comprehensive performance for example, local safety, local short-term efficacy, or/and medium and long-term efficacy
  • the object of the present invention is to provide a newly discovered topical activity based on immunoglobulins (hereinafter referred to as Ig) - that is, to convert one or more conventional ineffective drugs into their short-term synergistic, topical Synergistic safety, or/and mid- and long-term synergistic effects, the prepared topical pharmaceutical composition (hereinafter referred to as pharmaceutical composition) achieves high efficiency and low toxicity, especially in the medium and long term, providing various options for clinical use.
  • Ig immunoglobulins
  • immunoglobulin as a local active ingredient that can provide local synergy in the preparation of a pharmaceutical composition that can cause tissue necrosis in a target area, wherein the pharmaceutical composition further comprises the immune A topical synergist for globulin, and a suitable pharmaceutically acceptable carrier.
  • a pharmaceutical composition comprising an immunoglobulin that can provide a local synergistic effect, a local synergist of the immunoglobulin, and a suitable pharmaceutically acceptable carrier.
  • a method of treating or ameliorating local pathological diseases comprising the administration of a pharmaceutical composition that can cause tissue necrosis in a target area, wherein the pharmaceutical composition comprises a local synergistic effect.
  • Immunoglobulins, topical synergists of the immunoglobulins, and suitable pharmaceutically acceptable carriers comprising the administration of a pharmaceutical composition that can cause tissue necrosis in a target area, wherein the pharmaceutical composition comprises a local synergistic effect.
  • Embodiments according to the present invention have the following advantages over prior art pharmaceutical compositions comprising immunoglobulins: effective necrosis, or even ablation, of target tissue can be provided, while also potentially preserving the immunity (modulation) of immunoglobulins effect and provide higher local safety and/or better mid- and long-term efficacy.
  • Embodiments according to the present invention have the following advantages over other prior art for focal disease treatment: show less stringent screening of indications (can target pathogens in focal disease) compared to existing molecularly targeted drugs fibroblasts or/and pan-solid tumor cells and their associated interstitial cells), and great potential, for example, against rapidly growing tumors, large tumors, and hypovascular tumors; with existing locally active drugs such as chemical ablative agents Compared with conventional chemical ablative agents, it shows higher efficacy (especially the medium and long-term effect that conventional chemical ablation agents do not have) and significantly lower local irritation and damage to surrounding normal tissues, so that a wider range of interventional adaptations can be achieved. and higher application volumes.
  • the applications and compositions of the present invention are also not plagued by drug resistance problems encountered with existing cytotoxic drugs and existing molecularly targeted drugs.
  • the application and composition are convenient to prepare and low cost, and are particularly helpful for making safe and effective treatment available to a large number of people who cannot afford high costs.
  • the inventors of the present invention unexpectedly found in a tumor-bearing nude mouse experiment that is usually used as a chemotherapy model, although immunoglobulin (Ig) alone cannot provide local effects, but can be combined with some preferred substances under certain specific conditions, Local synergy is produced to effectively treat and even ablate tumor tissue. Further studies have also shown that mid- to long-term synergy can be produced to reduce the mid- to long-term rebound, or/and local synergistic safety that topical drugs typically have. These specific conditions are not constituents in conventional compositions of Ig, but are as defined below.
  • immunoglobulin as a synergist that can provide local synergy in the preparation of a pharmaceutical composition that can cause tissue necrosis in a target area, wherein the pharmaceutical composition further comprises the immune A topical synergist for globulin, and a suitable pharmaceutically acceptable carrier.
  • composition comprising an immunoglobulin that can provide local synergy, a local synergist of the immunoglobulin, and a suitable pharmaceutically acceptable carrier.
  • a method of treating or ameliorating a local pathological disease comprising the administration of a pharmaceutical composition that can cause tissue necrosis in a target area, wherein the pharmaceutical composition comprises a composition that can provide a local synergistic effect of immunoglobulins, topical synergists of said immunoglobulins, and suitable pharmaceutically acceptable carriers.
  • a device for the treatment of a localized disease comprising a topical pharmaceutical composition comprising immunoglobulin as defined in the present disclosure.
  • immunoglobulin is used as a structural concept beyond its conventional function in the prior art, and refers to all immunoglobulins that are identical or similar in composition to the immunoglobulins used in the examples of the present application , and can provide the locally active polypeptides disclosed in this application, and these polypeptides have only been found and used in the prior art for their immune (modulating) effects.
  • Immunoglobulins include natural immunoglobulins and their engineered analogs.
  • the natural immunoglobulin includes immunoglobulin G (IgG), immunoglobulin A (IgA), immunoglobulin M (IgM), immunoglobulin D (IgD), immunoglobulin E (IgE), usually from Obtained by separation of animal plasma, such as intravenous immunoglobulin G (IgG), intravenous immunoglobulin A (IgA), intravenous immunoglobulin M (IgM) and the like.
  • the said engineered analogs of natural immunoglobulins refer to engineered products (including genetically engineered products) that are different from natural immunoglobulins but have all or part of the key structures of natural immunoglobulins, such as immunoglobulin complexes, The following immunoglobulin fragments and their complexes (immunoglobulin Fc fragment, immunoglobulin Fab fragment, immunoglobulin Fab' fragment, immunoglobulin F (ab') fragment, single-linked immunoglobulin), and the like.
  • target volume is used to refer to the target site of administration, eg, adjacent to a localized lesion, interface, interior (preferably interior), and the like.
  • target tissue refers to the target tissue for administration, ie, a locally diseased tissue, eg, a locally diseased tissue containing cancer cells or/and fibroblasts.
  • the target when the lesion is a tumor, the target is the tumor, and the target area is the tumor area (for example, within the tumor or/and the periphery of the tumor where the drug can enter); when the lesion is non-tumor, the target is the tumor, and the target area is the tumor Large areas (such as hyperplasia, cysts, nodules, etc., or/and the surrounding area that can enter); when the lesion is localized inflammation, the target area is the inflamed area (such as an inflamed mass or/and the surrounding area where drugs can enter); When the lesions are abnormal secretory tissues, the target is the secretory glands with abnormal secretion, and the target area is the tissues or organs containing these abnormal secretory glands or/and their surroundings.
  • the pharmaceutical composition is preferably in a liquid dosage form, and the carrier includes water.
  • the immunoglobulins include polyclonal antibodies, preferably human immunoglobulins and engineered analogs thereof, more preferably human IgG and engineered analogs thereof.
  • the immunoglobulin is human IgG (including intravenous immunoglobulin, intramuscular immunoglobulin) or an engineered analog thereof (including genetically engineered IgG).
  • the immunoglobulin is selected from immunoglobulins other than anti-tumor monoclonal antibodies in the prior art, wherein the anti-tumor monoclonal antibodies refer to those used alone in the prior art, which are combined with Anti-tumor-specific immunoglobulins that can effectively treat tumors are shared with conventional chemotherapeutic drugs and with radiotherapy.
  • the local activity provided by the immunoglobulin includes forming at least one of the following local synergistic effects through the sharing of its local synergist under certain specific conditions: short-term local synergy, medium Long-term local synergy, local irritant antagonism.
  • the specific conditions shared by the immunoglobulin and its local synergist include a specific common reactor (the target tissue) and a specific concentration entering the common reactor (see the following immunoglobulin concentrations and its The shared amount of topical synergist such as weight ratio).
  • local synergists of immunoglobulins are sometimes referred to simply as immunoglobulin synergists.
  • the pharmacological effects of the pharmaceutical compositions disclosed in the present application include local synergistic effects (eg, short-term synergy or/and mid- and long-term synergies produced by topical co-administration) and other optional pharmacological effects in the target tissue.
  • local synergistic effects eg, short-term synergy or/and mid- and long-term synergies produced by topical co-administration
  • other optional pharmacological effects in the target tissue include local synergistic effects (eg, short-term synergy or/and mid- and long-term synergies produced by topical co-administration) and other optional pharmacological effects in the target tissue.
  • the synergy of the Ig and its conjugates is not that provided by the combination of their conventional antipathogenic activities (eg, cytotoxic, viral or bacterial activity), or any of their The synergy provided by the combination of conventional activities (eg, immunomodulatory and cytotoxic) is primarily the local synergy provided by the combination of their local activities.
  • the synergistic effect includes local chemical synergy (shown as a short-term synergistic effect) and optionally other pharmacological effects.
  • the synergistic effect includes mid- to long-term synergy (shown as mid- to long-term synergistic efficacy) and optionally other pharmacological effects.
  • the mid- to long-term synergy includes secondary immune synergy of the local chemical synergy and optionally other immune effects.
  • the pharmaceutical composition is in a topical dosage form.
  • topical dosage forms are distinguished from conventional dosage forms, which refer to dosage forms based on conventional activities and whose pharmaceutical composition is also suitable for systemic administration, so that systemic administration is predominant;
  • the former refers to a pharmaceutical dosage form that is based on local activity and whose drug composition is also suitable for the environment of the target area, so that it can only be delivered directly to the target area by interventional means.
  • Different dosage forms usually lead to different drug composition constraints, for example, topical dosage forms usually have to exclude pH adjusters (to make the drug as neutral as possible), osmotic pressure adjusters (to make the drug as close to the physiological osmotic pressure as possible), which are usually included in conventional dosage forms. , diluent (usually using pH and osmotic pressure as close to physiological conditions as possible), etc., because the excipients (such as NaCl) usually included in these conventional dosage forms may interfere with the local activity in the topical dosage form.
  • the pharmaceutical composition is a dosage form for local intralesional administration.
  • topical activity is distinguished from “conventional activity”, which refers to those obtainable by conventional administration (eg, intravenous injection, intracavity injection, subcutaneous administration, mucosal administration, etc.).
  • Activity such as cytotoxicity, immunity, etc.
  • Local activity includes, for example, the local action of a single drug and /or local co-occurrence of shared drugs.
  • Conventional activity can be achieved through the absorption of the drug through the blood after conventional medication to form drug-carrying blood, and then stimulate the immune system or enter the target area to produce conventional pharmacological effects (or conventional combined effects).
  • Local activity can only be achieved through local target area administration. Pharmacological effects (or co-actions) are produced, including chemical effects (or co-chemical effects) that are preferentially directed against the target tissue.
  • the term "local chemistry (or local shared chemistry)” refers to a chemical action (or shared chemistry) that occurs on a drug preferentially within a local tissue rather than a pathogen in that tissue or within the body's immune system, which appears to be short-term Efficacy.
  • topical active ingredient refers to a pharmaceutical ingredient which, when used alone or in combination, provides pharmaceutically activity including effective topical activity.
  • co-action refers to the co-pharmacological (or pharmacodynamic) action or co-toxicological (or co-toxicological (or side effects), which are often highly uncertain. If the joint action is the sum of the individual actions of each component, it is called “additive action”; if the joint action is less than the sum of the individual actions of each component, it is called “antagonism”: if the joint action exceeds the sum of the individual components The sum of the single effects is called “synergistic effect", for example, the effect of producing the actual/expected ratio q>1.15 of the medicinal effect agreed in the examples of the present invention.
  • local synergy refers to the synergy formed by local use, especially in local lesions, including short-term synergy based on short-term efficacy and mid- and long-term synergy based on mid- and long-term efficacy.
  • topical synergist refers to a pharmaceutical component that produces a local synergistic effect by topical administration.
  • therapeutically effective amount refers to a dose of a drug that is used to treat a disease (eg, tumor) and achieve an effective effect (eg, reduce or/and alleviate symptoms of the disease).
  • compositions disclosed herein are prepared, composed or administered such that the conditions required for the Ig to provide the local activity, or the local synergistic effect, are achieved by causing tissue necrosis, preferably effective necrosis, in the target area. More preferably, necrosis with minimal rebound in the medium to long term is used to treat localized disease.
  • tissue necrosis includes direct tissue necrosis and optionally indirect tissue necrosis.
  • the direct necrosis of the tissue is so severe that it is histochemically ablated.
  • the term "indirect necrosis of tissue” is used to refer to the effect of the inhibition of the proliferation of pathogens (eg cells, viruses or bacteria) in the target tissue or the stimulation of the immune system of the individual to the tissue
  • pathogens eg cells, viruses or bacteria
  • the former such as cytotoxic drugs
  • the latter such as anti-tumor vaccines, by stimulating the immune system to attack the cancer cells, may eventually lead to the necrosis of the tissue where the cancer cells are located.
  • Necrosis can then be achieved either by systemic or local administration.
  • tissue necrosis refers to tissue necrosis that is independent of indirect tissue necrosis caused by the direct application of a tissue disrupting factor to the target volume, wherein the tissue disrupting factor includes, for example: ablation components of a physical ablation instrument (e.g., warming needles) , conventional chemical ablative agents, etc.
  • a tissue disrupting factor includes, for example: ablation components of a physical ablation instrument (e.g., warming needles) , conventional chemical ablative agents, etc.
  • the term "ablation” means that the local effect of the in situ treatment is so strong that the target tissue produces tissue necrosis that essentially loses its original physiological function (inactivation).
  • Tissue necrosis includes coagulation necrosis, liquefaction necrosis, fibrinoid necrosis, and the like.
  • Tumor tissue necrosis can cause morphological changes (volume reduction) of tumor tissue, which ultimately manifests as an increase in tumor inhibition rate.
  • chemical ablation refers to ablation that is primarily chemical rather than immunological.
  • the term “chemical ablative agent” refers to a drug that exhibits an ablative effect only when administered topically.
  • concentration refers to the weight-volume percent concentration (w/v) of the specified component in the topical pharmaceutical composition.
  • local administration concentration refers to the concentration of a given component at the time of local administration of the drug, which may be the concentration of the given component at the target area of drug contact (eg, injection needle hole or perfusion tube outlet).
  • the pharmaceutical composition is prepared, composed or administered such that the immunoglobulin provides the local activity required by the immunoglobulin at a concentration (w/v) of ⁇ 2.0 , 2.0-35%, preferably 3.0-30%, eg, 4% to 30%, 4-20%, 3%-10%, 3%-8%, 4%-10%, into the target area.
  • the concentration (or amount ratio) in the term “entering the target area at a certain concentration (or amount ratio)" (or the concentration or amount ratio entering the target area) refers to the concentration (or amount ratio) at which the active ingredient begins to contact the target area, and this concentration (or amount ratio) is equivalent to the administration concentration (or administration concentration) in the local (target area) administration of the present invention.
  • dose ratio which is not equivalent to the dosed concentration (or dose ratio) in conventional (systemic) administration.
  • Quantitative ratios such as weight ratios.
  • the conditions required for the immunoglobulin to provide the local activity further include: the local synergist is selected from one or more of conventional ineffective drugs for the target region, and the immune
  • the globulin and the local synergist enter the target area with the following amount ratio (weight ratio) (W local synergist /W immunoglobulin ):
  • W local synergist /W immunoglobulin is ⁇ (0.5-39)/ 2.0, (0.5-39)/(2.0-35), or (1-39)/(2.0-35), preferably (3-30)/2.0, more preferably (4-30)/(2.0-35 ) or (5-20)/(4.0-30).
  • the immunoglobulin topical synergist comprises or is selected from one or more of conventional ineffective drugs for the localized disease.
  • the conventional ineffective drugs include or are selected from anti-tumor conventional ineffective drugs.
  • the conventional ineffective drugs include or are selected from anti-tumor or antiparasitic conventional ineffective drugs.
  • the conventional ineffective drugs include or are selected from conventional ineffective drugs against tumors or systemic pathogens (parasitic, bacterial, viral) infections.
  • the term "conventional ineffective compound (or drug)” is used to distinguish it from conventional effective compounds (eg, anti-tumor compounds), and means that in pathogen assays (eg, cell assays), it may show an effect against a specific pathogen ( For example, anti-tumor cell effect), but in animal experiments through absorption (conventional administration) does not show that the compound can achieve this anti-pathogen effect to effectively inhibit local disease disease (such as solid tumor).
  • Conventional ineffective compounds are not sufficiently active to be approved by pharmaceutical authorities (such as FDA) as conventional therapeutic drugs (such as antineoplastic drugs) for specific local diseased diseases.
  • the conditions required for the topical synergy further include: the topical synergist is one or more selected from the group consisting of amino acid nutrients, acid ablating agents, vital dyes, polysaccharides, and the Topical synergist enters the target area at the following dosing concentrations (weight-volume percent) (W topical synergist /V composition ): W topical synergist /V composition can be >0.35%-40%, >0.35% -39%, ⁇ 0.5%-40%, ⁇ 0.5%-39%, 0.5%-40%, 0.5%-39%, 1%-40%, or 1%-39%. , preferably (3-30)/2.0, more preferably (4-30)/(2.0-35) or (5-20)/(4.0-30).
  • amino acid nutrients refers to pharmaceutically acceptable amino acid compounds with nutritional health effects, preferably selected from amino acids, amino acid polymers and amino acid derivatives with nutritional health effects, more preferably It is selected from the amino acid nutraceuticals and amino acid excipients with nutritional and health care functions contained in the official pharmacopoeia or guideline of China, the United States or Europe.
  • the term “nutraceutical effect” is used to refer to an in vivo effect resulting from one or more of the following biological actions: providing energy, participating in the synthesis of biologically active substances (eg proteins), participating in partial metabolism, maintaining Animal intestinal microecological balance, and participate in other physiological regulation beneficial to body health (eg regulation of protein synthesis, regulation of immune response).
  • amino acids, amino acid polymers and amino acid derivatives as the amino acid-based nutrients are preferably amino acids selected from the group consisting of amino acids, or amino acid polymers containing amino acids from the group below, or amino acids selected from the group below.
  • Amino acid derivatives of amino acids or amino acid polymers comprising amino acids from the group of protein amino acids and non-protein amino acids.
  • protein amino acid refers to the main amino acids that make up protein
  • non-protein amino acid refers to protein amino acids other than protein amino acids, which can also be found in nutraceuticals, traditional diets and functional diets (for example, other amino acids used as nutritional health functional components in health diets).
  • the protein amino acids include amino acids selected from the group consisting of non-polar amino acids (eg, 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 (eg aspartic acid, glutamic acid). All of the above except glycine are L-type ⁇ -amino acids.
  • non-polar amino acids eg, alanine, valine, leucine, isoleucine, phenylalanine
  • proline polar neutral amino acids
  • polar neutral amino acids such as tryptophan, tyrosine, serine, cysteine, methionine, asparagine, glutamine, threonine
  • basic amino acids such as
  • the non-protein amino acids may include the following amino acids: ⁇ -alanine, taurine, ⁇ -aminobutyric acid (GABA), tea polyphenols (theanine), pumpkin seed amino acid (3-amino-3-carboxypyridine) alkanoic acid), glutamine, citrulline, ornithine, etc.
  • vitamin dye refers to a pharmaceutically acceptable (and thus already used in the human body) aromatic compound capable of selectively absorbing or reflecting light of a specific wavelength at a target area, which can Examples include methylene blue vital dyes.
  • the term "acid-ablating agent” is used to distinguish it from acid-base modulators, and means that topical administration at the concentration of the composition of the present invention exhibits a chemical ablative effect (eg, tumor-bearing animals have a tumor inhibition rate of 25%). % or more) acidic compounds, and these acidic compounds are not used as chemical ablation agents at lower concentrations, but as acid-base regulators with properties such as pH.
  • the acid ablative agent is a single drug that can show chemical ablation effect (for example, the tumor inhibition rate of tumor-bearing animals is more than 25%) at the concentration used, while the conventional drug is not provided
  • the conventional drug is not provided
  • no special biologically active acidic compounds are usually introduced.
  • the topical pharmaceutical composition comprises immunoglobulins, wherein the immunoglobulin concentration (w/v) may be ⁇ 2.0, 2.0-35%, preferably 3.0-30%, more preferably 4 % to 30%, even more preferably 4-20%, more preferably such as 3%-10%, 4%-10%, 3%-8%, 4%-8%.
  • the topical pharmaceutical composition comprises one or more amino acid nutrients.
  • the topical pharmaceutical composition comprises one or more vital dyes.
  • the topical pharmaceutical composition comprises one or more acid ablating agents.
  • the topical pharmaceutical composition comprises immunoglobulin, one or more amino acid based nutrients, and optionally one or more acid ablative agents.
  • the administration concentration (w/v) of the immunoglobulin in the pharmaceutical composition may be ⁇ 2.0, 2.0-35%, preferably 3.0-30%, more preferably 4% to 30%, more preferably such as 4-20%, 3%-10%, 3%-8%, 4%-10%.
  • the topical pharmaceutical composition comprises one or more amino acid nutrients and one or more acid ablating agents.
  • the topical pharmaceutical composition comprises one or more amino acid nutrients and one or more vital dyes.
  • the topical pharmaceutical composition comprises one or more acid ablating agents and one or more vital dyes.
  • the topical pharmaceutical composition comprises one or more amino acid nutrients, one or more acid ablating agents, and one or more vital dyes.
  • the administration concentration (w/v) of the amino acid nutrients in the pharmaceutical composition may be >7%, ⁇ 8%, preferably 8-30%, more preferably 10% -30% or 10%-20%, most preferably 7.5%-15%.
  • the administration concentration (w/v) of the vital dye in the pharmaceutical composition may be >0.3%, ⁇ 0.35%, 0.5-30%, preferably 0.35-10% or 1 %-5%, more preferably 0.5-3% or 1%-3%.
  • the administration concentration (w/v) of the acid ablating agent in the pharmaceutical composition may be ⁇ 0.5%, preferably 1.25%-10%, such as 1.5%-10%, 2 %-10%, 2.5%-10%, 2%-7.5%, or 2.5%-5%.
  • the amino acid-based nutrients are selected from one or more of basic amino acid-based nutrients and/or non-basic amino acid-based nutrients.
  • the topical pharmaceutical composition comprises the amino acid nutrient, wherein the amino acid nutrient comprises one or more selected from the group consisting of the following basic amino acid nutrient: arginine, lysine, group amino acid.
  • the topical pharmaceutical composition comprises the amino acid nutrient, wherein the amino acid nutrient comprises one or more selected from the group of non-basic amino acid nutrients: neutral amino acids, acidic amino acids, Amino acid salts, wherein the neutral amino acids such as: glycine, tryptophan, tyrosine, serine, cysteine, methionine, asparagine, glutamine, threonine, alanine, valine, Leucine, isoleucine, phenylalanine, proline, the acidic amino acids such as aspartic acid, glutamic acid, and the amino acid salts include the salts formed by the above-mentioned amino acids and acids, such as Lysine hydrochloride, histidine hydrochloride, glutamic acid hydrochloride, cysteine hydrochloride, arginine hydrochloride, glycine sulfate, iron glycinate sulfate, lysine hydrochloride, Aspart
  • the topical pharmaceutical composition comprises the amino acid nutrient, wherein the amino acid nutrient comprises an amino acid or a salt thereof selected from the group consisting of or one of oligopeptides and polypeptides comprising or consisting of the following amino acids One or more: arginine, lysine, glycine, cysteine, alanine, serine, glutamic acid.
  • the oligopeptide may include glutathione.
  • the amino acid nutrient is arginine
  • the administration concentration (w/v) of the arginine in the pharmaceutical composition is ⁇ 10%, preferably 10-30%, more Preferably it is 15%-20%.
  • the topical pharmaceutical composition comprises the amino acid nutrient, wherein the amino acid nutrient comprises glycine.
  • the pharmaceutical composition comprises glycine, and the administration concentration (w/v) of the glycine in the pharmaceutical composition is ⁇ 10%, preferably 10-30%, more preferably 10% -15%.
  • the pharmaceutical composition comprises lysine, and the administration concentration (w/v) of the lysine in the pharmaceutical composition is ⁇ 10%, preferably 10-30%, more It is preferably 10-20%, such as 15%.
  • the topical pharmaceutical composition comprises a plurality of the amino acid nutrients, and the administration concentration (w/v) of the plurality of amino acid nutrients in the pharmaceutical composition may be ⁇ 10% , preferably can be 10-30%, more preferably can be 10-20%, such as 10%-15% or 15%-20%.
  • the topical pharmaceutical composition comprises the vital dye, wherein the vital dye is preferably one or more selected from the following compounds and derivatives thereof: methylene blue, patent blue, isosulfide blue.
  • the topical pharmaceutical composition comprises the vital dye, wherein the vital dye comprises methylene blue, and wherein the methylene blue is administered at a concentration (w/v) of 0.5% to 5.0 %, preferably can be 1%-5%, more preferably can be 1-3% or 1-2%, for example 1%, 2% or 3%.
  • the pharmaceutical composition comprises methylene blue and the amino acid nutrient.
  • the pharmaceutical composition comprises methylene blue and polysaccharide.
  • the polysaccharide concentration (w/v) is 1-20%, preferably 1.5-20%, more preferably 1.5-15%, such as 10%, or 15%.
  • the polysaccharide comprises or is selected from one or more of the following: dextran, such as carboxymethyl dextran and water-soluble beta-D-glucan, dextran.
  • dextran such as carboxymethyl dextran and water-soluble beta-D-glucan, dextran.
  • the concentration (w/v) of the carboxymethyl glucan may be 10%-15%, such as 10%; the concentration of the water-soluble ⁇ -D-glucan may be 10%- 15%, such as 15%; the concentration (w/v) of the dextran can be 1.5%-10%, such as 1.5%.
  • the pharmaceutical composition comprises immunoglobulin, methylene blue and optionally polysaccharide, wherein the polysaccharide concentration (w/v) may be 1-20%, preferably 1.5-20%, More preferably it may be 1.5-15%, such as 10%, or 15%.
  • the topical pharmaceutical composition comprises the acid ablating agent, wherein the acid ablating agent is selected from one or more of the following group: weak acid, strong acid.
  • the topical pharmaceutical composition comprises the acid ablating agent, wherein the acid ablating agent has a concentration in the pharmaceutical composition at which a single agent exhibits chemical ablative efficacy, such as the
  • the weak acid may be administered at a concentration (w/v) of ⁇ 2.0%, ⁇ 3.0% or ⁇ 5%, preferably 3.0-20% or 5-20%, more preferably 5-10%, such as 7.5% or 10%; the administration concentration (w/v) of the strong acid can be ⁇ 0.5%, ⁇ 0.75% or ⁇ 1%, preferably 0.5-3% or 0.5-3%, such as 1% or 2%.
  • the weak acid is preferably one or more selected from the group consisting of carbonic acid, acetic acid, glycolic acid, propionic acid, malonic acid, butyric acid, succinic acid, lactic acid (2-hydroxypropionic acid) , citric acid (2-hydroxy-1,2,3-propanetricarboxylic acid), malic acid (2-hydroxysuccinic acid), tartaric acid, oxalic acid, gluconic acid; preferably acetic acid.
  • the strong acid includes, for example, hydrochloric acid, sulfuric acid, nitric acid, perchloric acid, selenic acid, hydrobromic acid, hydroiodic acid, preferably hydrochloric acid.
  • the pharmaceutical composition comprises the amino acid nutrient and a weak acid or/and a strong acid, wherein the amino acid nutrient comprises glycine; the weak acid comprises acetic acid; and the strong acid comprises hydrochloric acid.
  • the pharmaceutical composition comprises the amino acid nutrient, weak acid and strong base salt or/and weak acid, wherein the amino acid nutrient comprises glycine; the weak acid comprises acetic acid; the weak acid and strong base salt comprises acetic acid sodium.
  • the pharmaceutical composition comprises the amino acid nutrient, a weak acid and strong base salt, and a vital dye, wherein the amino acid nutrient comprises glycine; the vital dye comprises methylene blue; the weak acid strong base salt Includes sodium acetate.
  • the pharmaceutical composition comprises the amino acid nutrient, a weak acid, and a vital dye, wherein the amino acid nutrient comprises glycine; the vital dye comprises methylene blue; and the weak acid comprises acetic acid.
  • the pharmaceutical composition disclosed according to the present application can be any dosage form suitable for topical administration that can contain the active ingredient (the pharmaceutical composition), preferably the following dosage forms: injection (preferably local injection), external solution, atomization agent, etc.
  • the pharmaceutical composition is a powder for injection.
  • the pharmaceutical composition is a liquid injection.
  • the term "injectable” is used to refer to a sterile preparation containing the active ingredient and a liquid carrier for in vivo administration.
  • the injections are classified into local injections, intravenous injections, etc. according to the mode of administration, and the intravenous injections can be used as local injections only after a given local administration concentration.
  • Injections are classified into liquid injections, powder injections, etc.
  • a powder for injection contains sterile dry powder and a vehicle, the sterile dry powder contains part or all of the active ingredients, and the vehicle 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, usually the end point (such as needle hole, catheter outlet, etc.) of the local drug delivery device (syringe, trocar, perfusion catheter, etc.) ) of the active ingredient concentration in the liquid medicine.
  • the concentration of the active ingredient is the concentration of the active ingredient in a mixture of sterile dry powder and a vehicle (eg, a reconstituted solution, or the pharmaceutically acceptable liquid carrier).
  • liquid for external use refers to an active ingredient and a liquid carrier and a donor surface [e.g. skin, mucous membranes (e.g. ocular mucosa, nasal mucosa, etc.) or/and cavities (e.g. oral cavity, rectum, Vaginal, urethral, nasal, ear canal, etc.)] administration of liquid medicines, which include, for example, lotions, liniments, drops, gargles, paints, and the like. In the case of topical administration, the liquid medicine often comes from local administration devices such as lotion bottles, drip bottles, drip tubes, rinse bottles, cotton swabs, and the like.
  • the concentration of the active ingredient in the external liquid preparation is the concentration of the active ingredient in the liquid medicine.
  • the term "nebulizer” refers to a dosage form which contains the active ingredient and a liquid carrier and which, in use, can be administered by atomizing the aforementioned liquid drug by means of pressure, which can be applied to the skin, mucous membranes (e.g. ocular mucosa, nasal mucosa, etc.) or/and cavity (eg, oral, rectal, vaginal, urethral, nasal, ear canal, etc.) administration, including, eg, aerosols, sprays, nebulizers, and the like.
  • the atomization of liquid drugs is often formed by topical drug delivery devices such as aerosols, atomizers, and nebulizers.
  • the drug After the drug is atomized and sprayed to the target position, it accumulates to form a liquid drug.
  • the liquid medicine is substantially the same in composition as the liquid medicine before atomization.
  • the concentration of the active ingredient in the nebulizer can be represented by the concentration of the active ingredient in the liquid drug prior to nebulization.
  • the pharmaceutical composition of the present invention should be made into a dosage form that can be locally administered to the target area.
  • the preparation of the pharmaceutical composition of the present invention comprises the steps of preparing a liquid medicine containing the essential components (such as the pharmaceutical composition) and optionally other substances.
  • the liquid drug can be a solution (eg, in a hydrophilic vehicle, preferably an aqueous solution), a suspension, or an emulsion.
  • 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 for injection.
  • the vegetable oil may be, for example, cottonseed oil, almond oil, olive oil, castor oil, sesame oil, soybean oil and peanut oil.
  • compositions disclosed in accordance with the present application can be prepared or composed such that the Ig or/and its co-conjugates can be contained in a concentration greater than or equal to its administration concentration required to satisfy the local synergistic effect. When it is greater than the administration concentration, it can be further diluted for use.
  • the liquid injection of the pharmaceutical composition of the present invention can be prepared by a method comprising the following steps: 1) The necessary components (eg, the pharmaceutical composition) in the required amount according to the local administration concentration And other components optionally present are added into water to prepare a liquid; 2) the other drugs optionally present according to the required amount of local administration concentration are added to the liquid prepared in 1) and mixed uniformly to obtain a liquid drug; 3) will be in the The liquid medicine prepared in 2) was sterilized to make a liquid injection. In use, the sterilized liquid medicine in the liquid injection can be used as a liquid medicine for topical administration directly or after dilution.
  • the injection powder of the pharmaceutical composition of the present invention can be prepared by a method comprising the following steps: preparing a sterile dry powder containing the pharmaceutical composition in the required amount according to the local administration concentration; and preparation of sterile vehicles containing the required amounts of the optional other components according to the concentration for topical administration.
  • Described sterilization dry powder is preferably sterilization freeze-dried dry powder, and its preparation method comprises: 1) prepare the solution that comprises amino acid to nutrient, water-soluble weak neutralization substance and optional other components; 2) sterilization filtration and separation; 3) freeze-drying; 4) plugging and capping.
  • the freeze-drying process conditions include, for example: the pre-freezing conditions are kept at the pre-freezing temperature -45°C for 4 hours; the sublimation drying conditions are that the heating 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 were kept at 30°C for 6 hours.
  • the sterile dry powder of the powder for injection is reconstituted in a sterile solvent to form a reconstituted liquid drug, which can be used directly or after dilution as a liquid drug for topical administration.
  • the external use liquid of the pharmaceutical composition of the present invention is prepared by a method comprising the following steps: adding the required amount of the pharmaceutical composition according to the local administration concentration and optionally other components into a vehicle It is prepared as a liquid medicine.
  • the liquid medicine in the external solution can be used directly or after being diluted as a liquid medicine for topical administration.
  • the pharmaceutical composition aerosol of the present invention can be prepared by a method comprising the following steps: 1) adding the required amount of the pharmaceutical composition according to the local administration concentration and aerosol excipients into The vehicle is prepared as a liquid; 2) the other components optionally present in the required amount according to the local administration concentration are added to the liquid prepared in 1) and mixed uniformly to obtain a liquid medicine.
  • Commonly used excipients for nebulization include, for example: glycerin, polysorbate-80, benzalkonium chloride, microcrystalline cellulose-sodium carboxymethyl cellulose, and the like.
  • the liquid drug is added to a nebulizer (eg, a spray) and is locally administered to the target area in the form of mist under its atomization, and the mist is accumulated in the target area as a liquid drug.
  • a device for treating local pathological diseases comprising the pharmaceutical composition according to the present disclosure.
  • the device comprises an aerosol, nebulizer, nebulizer.
  • compositions of the present invention include: containing different types and concentrations of the pharmaceutical composition, containing different types and concentrations of other drugs, containing different types and concentrations of other additives (eg, analgesics, activators, etc.) Wait).
  • other additives eg, analgesics, activators, etc.
  • the pharmaceutical composition is mainly used for the prevention and treatment of local pathological diseases, especially refractory local pathological diseases, by topical administration.
  • the term "focal disease” refers to a disease with symptoms of a local disease
  • the term “focal disease” refers to a structural, morphological or functional disorder that is native or secondary to a local part of the body of an animal (preferably a human).
  • Abnormal for example, may include one or more of the following: tumor, non-tumor swelling, local inflammation, abnormal secretion of secretory glands, and the like.
  • the local site can be any suitable one known to those skilled in the art, for example, it can be a local part including one or more of the following organs: the secretory organ where the secretory system is located, the cardiovascular organ where the blood circulatory system is located, the skin, etc. .
  • the drug composition (local active ingredient, composition ratio and component concentration) can be administered to the tissue where the local lesion is located by interventional means, and produce the desired therapeutic effect in the tissue.
  • the local tissue is the tumor body where the tumor cells are located; when the lesion is non-tumor, the local tissue is an abnormality such as a mass, such as hyperplasia, cyst, nodule, etc.; when the lesion is a local
  • the local tissue is the inflamed area, such as an inflamed body; when the lesion is abnormal secretion, the local tissue is the abnormal source or the secretory gland where it is located.
  • the abnormal source is the pancreatic islet
  • the local tissue is the pancreatic islet or the pancreas where the pancreatic islet is located
  • the disease is a skin disease
  • the local tissue is the diseased skin or an accessory organ of the diseased skin.
  • the target tissue is a local lesion tissue containing cancer cells or/and fibroblasts; the local lesion includes tumor, non-tumor swelling, local inflammation, abnormal secretion gland function and skin disease.
  • tumor refers to a mass formed due to abnormal proliferation of cells or mutated cells, including solid tumors.
  • solid tumor refers to a tumor with a tumor body, which can be due to any pathology (malignant and non-malignant) and at any stage, including, for example, the following groups classified by tumor cell type: epithelial cell tumors, sarcomas, Lymphoma, germ cell tumor, blastoma; and tumors named after the organ or tissue in which the tumor cell concentration is located, including, for example, tumors named after the following organs or tissues: skin, bone, muscle, breast, kidney, Liver, Lung, Gallbladder, Pancreas, Brain, Esophagus, Bladder, Large Intestine, Small Intestine, Spleen, Stomach, Prostate, Green Pill, Ovary or Uterus.
  • the malignant tumor includes, for example, breast cancer, pancreatic cancer, thyroid cancer, nasopharyngeal cancer, prostate cancer, liver cancer, lung cancer, colon cancer, oral cancer, esophageal cancer, gastric cancer, throat cancer, testicular cancer, vaginal cancer, Uterine cancer, ovarian cancer, etc.
  • the non-malignant tumors include, for example, breast tumors, pancreatic tumors, thyroid tumors, prostate tumors, liver tumors, lung tumors, intestinal tumors, oral tumors, esophageal tumors, gastric tumors, nasopharyngeal tumors, laryngeal tumors, testicular tumors, vaginal tumors, Uterine tumor, fallopian tube tumor, ovarian tumor, etc.
  • the locally diseased disease comprises a non-tumorous enlargement.
  • non-neoplastic refers to enlargements other than tumors, including, for example, hyperplasia (eg, hyperplasia of breast, pancreas, thyroid, parathyroid, prostate, etc.), cysts (eg, cysts of breast, thyroid, parathyroid, etc.) , Nodules (such as nodules of breast, thyroid, parathyroid, etc.), abnormal venous mass (such as hemorrhoids, etc.), local inflammation and swelling, microbial infection and swelling.
  • the hemorrhoids include internal hemorrhoids, external hemorrhoids and mixed hemorrhoids.
  • the locally diseased disease comprises local inflammation, especially refractory inflammation.
  • localized inflammation refers to non-neoplastic inflammation of a localized site, including, for example, alterative, exudative, and proliferative inflammation, which may be in the art Any suitable known to the skilled person, for example, may include one or more of the following: arthritis, mastitis, pancreatitis, thyroiditis, prostatitis, hepatitis, pneumonia, enteritis, stomatitis, pharyngitis, periodontitis, Esophagitis, gastritis, gastric ulcer, rhinitis, sinusitis, laryngitis, bronchitis, bronchitis, vaginitis, metritis, salpingitis, oophoritis, etc.
  • the localized disease comprises a skin disease, especially a refractory skin disease.
  • skin disease refers to a disease that is native or secondary to the skin or skin appendages, which may be any suitable known to those skilled in the art, for example may include one or more of the following Species: skin cancer, non-malignant skin tumors, viral skin diseases (e.g. herpes, warts, rubella, hand, foot and mouth disease), bacterial skin diseases (e.g. impetigo, boils, leprosy), fungal skin diseases (e.g.
  • various ringworm sexually transmitted diseases (such as syphilis, gonorrhea, and condyloma acuminatum), allergic and autoimmune skin diseases (such as contact dermatitis, eczema, urticaria), physical skin diseases (such as solar dermatosis, frostbite, corns) , chapped hands and feet, pressure ulcers), connective tissue diseases (such as lupus erythematosus), dyspigmented skin diseases (such as freckles, pigmented nevi, various macules), skin adnexal diseases (such as acne, rosacea, seborrheic dermatitis) , alopecia areata, alopecia, hyperhidrosis and stinkyhidrosis).
  • sexually transmitted diseases such as syphilis, gonorrhea, and condyloma acuminatum
  • allergic and autoimmune skin diseases such as contact dermatitis, eczema, urtic
  • the focal disease comprises secretory gland dysfunction.
  • secretory gland refers to a structure composed of glandular cells or groups of glandular cells that performs a secretory function, including exocrine and endocrine glands.
  • Abnormal secretion function of secretory glands includes hypersecretory gland function (eg, hyperthyroidism) and hyposecretory gland function (eg, hypothyroidism, islet dysfunction (a type of diabetes)).
  • the localized disease comprises cardiovascular disease.
  • Interventional therapy has become an important treatment for cardiovascular disease.
  • cardiovascular diseases include, for example, hemangiomas, hypertrophic obstructive cardiomyopathy, atrial fibrillation, arrhythmias, arterial embolism, and the like.
  • the topical drug disclosed in the present application is a therapeutic drug, and when it is used for the prevention and treatment of local lesions, it can also be combined with other interventional therapy, systemic chemotherapy, immunotherapy, photodynamic therapy, sonodynamic therapy, surgical intervention or Combinations of such therapies are administered in combination to further enhance efficacy.
  • the pharmaceutical composition is mainly used for the prevention and treatment of local pathological diseases through topical administration.
  • the one comprising immunoglobulin and its topical synergist is administered locally in the concentration or amount ratio thereof in the topical pharmaceutical composition .
  • This concentration or amount ratio can provide a synergistic effect of the local response when administered locally.
  • the method comprises optionally performing one or more other treatments, eg, chemotherapy, immunotherapy, radiation therapy, surgery, physical ablation, before, during, or after topical administration of the pharmaceutical composition.
  • treatments eg, chemotherapy, immunotherapy, radiation therapy, surgery, physical ablation, before, during, or after topical administration of the pharmaceutical composition.
  • the pharmaceutical composition of the present invention is shown to promote the promotion of local lesions in the relevant structures of the tissue (eg, diseased tissue, diseased cells, and any structure involved in constituting them). It can effectively destroy and at the same time only minimize the damage to the normal tissue of patients, so as to achieve the pharmaceutical effect of safe and effective treatment of local lesions.
  • tissue eg, diseased tissue, diseased cells, and any structure involved in constituting them.
  • mice There are two types: BALB/c mice and nude mice. Nude mice are mutant (BALB/c–nu) mice obtained by introducing nude gene (nu) into BALB/c mice. The mice were all healthy females aged 6-8 weeks, weighing 17.5-20.5 g.
  • Tumor volume V l/2 ⁇ a ⁇ b 2 , where a represents tumor length and b represents tumor width.
  • Volume tumor inhibition rate r(%) (CV-TV)/CV ⁇ 100%, where TV is the average volume of the study group; CV is the average volume of the negative control group.
  • 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.
  • the efficacy of drug i is denoted as E i , where E i can be (100-r i )% or R i %.
  • drug 1/drug 2/.../drug n the composition of drug A and drug B
  • B/A the composition of drug A and drug B
  • the single drug efficacy of A and B is recorded as EA and EB respectively, and the actual combined efficacy of A/B is recorded as EA +B .
  • test results (such as tumor weight, tumor volume) are expressed as mean ⁇ standard deviation (x ⁇ s), and the difference between the two experimental animal groups and the group mean is expressed by statistical software SPSS 13.0 or SPSS 19.0 software was used to carry out a significant test for comparison, and the test was carried out with the statistic t.
  • the type of drug action (common pharmacology) is studied by drug effect, especially comparing the drug effect of the same investigational drug in different technical protocols.
  • the difference in drug effect of drug i between regimens X and Y is not uncommonly large (eg, E iX /E iY ⁇ 150%), it is likely to be different kinetics in essentially the same type of drug action (pharmacology) and when the drug effect difference is unusually large (eg E iX /E iY >150%), the drug effect of drug i in regimen X should be greater than its The expected range of kinetics for the type of drug action (pharmacology) in Y, and thus likely to involve a different type of drug action (pharmacology) than in regimen Y.
  • systemic active positive controls include classical cytotoxic drugs (such as 0.5-1% 5-fluorouracil, whose tumor inhibition rate is ⁇ 30% under the conditions of the following examples), and the locally active positive controls include classical Chemical ablative agents (eg, 75-99% ethanol, whose tumor inhibition rate is ⁇ 30% under the local administration conditions of the following examples).
  • classical cytotoxic drugs such as 0.5-1% 5-fluorouracil, whose tumor inhibition rate is ⁇ 30% under the conditions of the following examples
  • locally active positive controls include classical Chemical ablative agents (eg, 75-99% ethanol, whose tumor inhibition rate is ⁇ 30% under the local administration conditions of the following examples).
  • EA /B is the actual combined efficacy (such as tumor inhibition rate) of composition A/B
  • EA and EB are the actual drug efficacy of component A and component B of composition A/ B , respectively
  • the combined pharmacodynamic effect of the combined administration of drugs A and B according to the above-mentioned Kim Jong-jun method is judged as follows: when the drug efficacy of the combination of drugs A and B has no therapeutic significance (for example, r ⁇ 30% ), the composition showed no synergy, or considered negligible synergy.
  • the combined efficacy of the composition has a significant synergistic effect, that is, the actual effect exceeds the theoretical purely additive expectation; if the actual efficacy/expected efficacy ratio q ⁇ 1.00 Then it shows that the combined efficacy of the composition has obvious antagonistic effect, that is, the actual effect is not as expected by the simple addition of the theory.
  • the combined effect of combined administration is judged, that is: when the composition A/B group does not show a therapeutically significant effect (for example, r ⁇ 30%) ), the co-administration of A and B also does not show a therapeutically meaningful synergy, which is regarded as a negligible synergy in the present invention.
  • the combined effect of the composition was additive if the actual/expected ratio q was between 0.85 and 1.15 (the actual effect was in line with the theoretical pure phase) If the actual/expected ratio q>1.15, the synergistic effect of the composition is obviously synergistic (the actual effect exceeds the theoretical purely additive expectation), and if the actual/expected ratio q ⁇ 0.85, the synergistic effect of the composition is Obvious antagonistic effect (the actual effect is not as expected by the theoretical addition).
  • the necessary components eg, 5 g of human immunoglobulin, 13 g of glycine, and 10 g of acetic acid
  • optional other components eg, a liquid carrier (eg, 100 ml) of the pharmaceutical composition
  • a liquid carrier eg, 100 ml
  • the preparation eg, 5% human immunoglobulin/13% glycine/10% acetic acid in water
  • the necessary components eg, 5 g of human immunoglobulin, 1 g of methylene blue), optional other components, and a liquid carrier (eg, 100 ml) of the pharmaceutical composition are dosed to the desired concentration in the pharmaceutical composition Water for injection), slowly mix them evenly, and after sterilization filtration, pack into the required amount (for example, 10ml/bottle), freeze-dry, cork, and cap, and prepare as sterilized dry powder for later use.
  • the required amount of sterile dry powder (for example, 1 bottle of the above dry powder) is reconstituted in the required amount of sterile liquid (for example, water for injection) to obtain the desired reconstituted solution (for example, 5% human immunoglobulin). /1% methylene blue aqueous solution) can be administered locally as a liquid drug.
  • the necessary components eg, 5 g of human immunoglobulin, 13 g of glycine, and 10 g of acetic acid
  • a liquid carrier eg, 100 ml
  • the preparation eg, 5% human immunoglobulin/13% glycine/10% acetic acid in water
  • the necessary components in the pharmaceutical composition eg 3g or 5g human immunoglobulin and 13g glycine and 10g acetic acid
  • the desired concentration as described in Table 1
  • aerosol excipients glycerol (2.5g), Polysorbate-80 (1.5 g), benzalkonium chloride (0.02 g), microcrystalline cellulose-sodium carboxymethyl cellulose (1.5 g)
  • a vehicle e.g. water for injection
  • the preparation can be used as a spray stock solution, and can be directly sprayed on the target area to form a liquid medicine after being added to the sprayer.
  • immunoglobulins in the prior art in the preparation of tumor therapeutic drugs is entirely based on their conventional activities (eg, immunomodulatory pharmacology), and has a narrow application range, and is mainly used as an adjuvant therapeutic drug rather than a therapeutic drug.
  • the development of new applications of immunoglobulin is essentially the development of new pharmacology of immunoglobulin.
  • the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1 ⁇ 10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model.
  • Successfully modeled experimental animals (average tumor volume 317 mm 3 ) were randomly divided into 2 negative control groups (01, 02) and 12 drug study groups (1-12).
  • the negative control is normal saline
  • the research drugs are shown in the table below
  • the human immunoglobulins are the freeze-dried intravenous human immunoglobulins in Table 1.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1.
  • Each experimental group was administered by systemic administration (intravenous injection), local administration in the target area (intratumoral injection), and mixed administration (intravenous injection + intratumoral injection) as shown in the table below.
  • Each group was administered once every 3 days, a total of 3 times, and the injection volume was 150 ⁇ l/only/time.
  • the animals were euthanized 3 days after the drug administration, the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 2.
  • the intravenous injection group (group 4) and the intratumoral injection group (group 9) of the cytotoxic drug (5-fluorouracil) showed almost indistinguishable tumor inhibition rates, although it has been suggested that intratumoral administration may improve the The local drug concentration will thus significantly increase its efficacy.
  • the intravenous injection group (group 4) and the intratumoral injection group (group 9) of the chemoablation agent (ethanol) showed a significant difference (>150%) in tumor inhibition rate, in line with its two dosing regimens Different pharmacological responses are expected.
  • the pharmacology of systemic administration of ethanol is consistent with that of drinking alcohol, while the pharmacology of local administration of ethanol is that local chemical action can cause damage to diseased tissue.
  • immunoglobulins achieve their pharmacology as immunomodulators by targeting immune system molecules, and this targeting determines their clinical use as a form of routine administration (e.g. intravenous, intracavitary, subcutaneous administration) , mucosal administration, intramuscular injection, etc.)-based anti-tumor adjuvant drugs.
  • routine administration e.g. intravenous, intracavitary, subcutaneous administration
  • mucosal administration e.g. intramuscular injection, etc.
  • intramuscular injection e.g. intramuscular injection
  • topical administration group 6 were consistent with routine administration group 1, indicating that the local application of any conventional activity of the immunoglobulin, or its single-agent local activity, could not produce tumor growth inhibition.
  • immunoglobulin (X) has only conventional activities, and its combination (X/Y) with any drug (Y) can only provide these conventional activities for combined action when it is administered systemically (for example, The combination of X immunomodulatory effect and Y conventional anti-tumor effect, or/and the combination between X’s conventional anti-tumor effect and Y’s conventional anti-tumor effect), can only provide these conventional activities for combined effect in topical administration (e.g. Combination of X immunomodulatory effect and Y conventional antitumor effect or local effect).
  • the difference in tumor weight was statistically significant (p ⁇ 0.05), and the difference in tumor inhibition rate was as high as 10 times or more, indicating that the drug composition in group 8 was different from group 3.
  • the new pharmacology realizes tumor tissue ablation.
  • the tumor inhibition rate of composition group 8 also exceeded the expected range of theoretical pure additive effect, and its actual/expected ratio (q) was greater than 1.15, indicating that the above-mentioned new pharmacology should be a local synergistic pharmacology.
  • the activity provided by the immunoglobulin in this local synergistic pharmacology is clearly beyond the expectations of any conventional activity and is a local activity.
  • the drug target is one of the keys to the pharmacology and therefore the efficacy of the drug.
  • the completely different pharmacological effects of the immunoglobulin-containing compositions in the above experiments are likely to be due to their completely different drug targets and pharmacology.
  • Conventional compositions enter the body through conventional drugs, mainly distributed in the blood and de-targeted in the form of blood drugs (eg study group 3), the target can be the immune system, cancer cells, and so on.
  • the topical composition enters the body through local administration, mainly relying on short-term external force to distribute and penetrate in the gap in the local lesion (tumor tissue) (for example, research group 8), which acts on the tissue first, and then in it. Contents (cancer cells, immune cells).
  • the immunomodulatory action of immunoglobulin is denoted as A1, the characteristic composition required for this action is denoted as X1; its conventional antitumor action (if any) is denoted as A2, and the required characteristic composition for this action is denoted as A2.
  • the characteristic composition is denoted X2; its local activity (effective local synergy) is denoted A3, and the characteristic composition required for this activity is denoted X3 (such as those described in the above and following examples). From a pharmaceutical rather than chemical point of view, these are three immunoglobulins (A1X1, A2X2) that provide different activities (A1, A2, A3) and also have different structure-effect profiles (X1, X2, X3) , A3X3).
  • A1X1 and A2X2 should be shown in systemically administered animals in the above experiments if there is a significant, not to say, effective drug effect.
  • A3X3 can prepare the composition of the present invention which far exceeds the expected technical effect (dosing efficiency) of A1X1 and A2X2.
  • the functional combination or composition of the composition of the present invention is not 1): a combination of immunoglobulin immune effect and co-use local effect (B1) (A1/B1), or A1X1/B1Y Composition, nor 2): a combination (A2/B1) of the conventional antitumor effect of immunoglobulin (if any) (A2) and the local effect of the concomitant (B1), or an A2X2/B1Y composition, but 3): Local activity of immunoglobulin (local synergy, A3) and combination of local synergy (B2) of immunoglobulin (A3/B2), or A3X3/B2Y composition.
  • the experimental animals were nude mice, the model cells were sarcoma S180, and 100ul (1 ⁇ 10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animals.
  • Successfully modeled experimental animals (sarcoma-bearing mice, with an average tumor volume of 226 mm 3 ) were randomly divided into 8 experimental groups (1 negative control group and 7 research groups).
  • the negative control is normal saline
  • the research drugs are shown in the following table
  • the human immunoglobulin is the recombinant human IgG in Table 1.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1. Each group was administered once every 3 days for a total of 3 times.
  • the animals were euthanized 3 days after the drug administration, and the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group.
  • the results are shown in Table 3.
  • Nude mice are congenital athymic nude mice obtained by introducing a recessive mutation gene "nu" on the 11th pair of chromosomes into BALB/c mice.
  • the thymus has only remnants or abnormal epithelium, which cannot make T cells differentiate normally, lacks the auxiliary, inhibitory and killing functions of mature T cells, and has low immunity. It has a high degree of immune tolerance to xenogeneic biological substances, for example, there is almost no transplant immune response to xenogeneic cancer cells.
  • tumor-bearing nude mice are widely used as models of chemotherapy rather than immunotherapy.
  • immunoglobulins (even xenogeneic immunoglobulins) as immunomodulatory drugs should not exhibit immunosuppressive, let alone local effects, let alone local effects in tumor-bearing nude mice experiments chemical action.
  • the results of Study Group 1 met this expectation.
  • composition groups 3, 5, 7 and group 1 were statistically significant (P ⁇ 0.05), and their respective actual/expected ratios (q) were all greater than 1.15, all showing short-term synergy. .
  • This short-term synergy should include local chemical synergy.
  • Nude mice were used as experimental animals, and human hepatoma HepG2 cells were used as transplanted tumor model cells. According to the well-known method, 1 ⁇ 10 5 cancer cells were subcutaneously injected into the right armpit of each animal. When the transplanted tumor grew to an average volume of 127 mm, the tumor-bearing animals were randomly divided into 3 series A, B, and C, and each series was randomly divided into 5 groups (group 0, 1, 2, 3, and 4).
  • group 0 is a negative control group, given normal saline
  • group 1 is a chemical ablation control group, given anhydrous ethanol
  • group 2 is a shared substance group, given 20% glycine/10% acetic acid
  • Group 3 is an immunoglobulin group, given 4% human immunoglobulin
  • group 4 is a composition group, given 4% human immunoglobulin/20% glycine/10% acetic acid.
  • the medicines are all aqueous solutions, and the research medicines are prepared according to the preparation method of Example 1, and the human immunoglobulin in it is the human IgG in Table 1.
  • mice were administered once every 2 days for a total of 3 times.
  • Each group in series A received intratumoral injection (75 ⁇ l/animal/time), each group in series C received tail vein injection (75 ⁇ l/animal/time), and each group in series B received intratumoral injection Injection (75 ⁇ l/animal/time) and tail vein injection (75 ⁇ l/animal/time), including group 4 intratumoral injection of 20% glycine/10% acetic acid and tail vein injection of 4% human immunoglobulin.
  • the tumor volume of animals in each series and each group was measured, and the 7-day volume tumor inhibition rate (r7) and The volume tumor inhibition rate (r21) on the 21st day, the results are shown in Table 4 below.
  • the immunoglobulin composition (group 4) has both r7 and r21 in series C ⁇ 5 %, indicating that the components of the composition can be used as any activity for conventional activities (including this cytotoxic activity) ) had no short-term and mid-term and long-term tumor inhibitory effects; in series B, its efficacy was similar to that of the common component (group 2), indicating that any activity (including immunomodulatory activity) that immunoglobulins can be used as conventional activities is comparable to The combined use of the topical activity of the common components did not exceed the short-term activity expectations (q 7 >1.00) and medium and long-term activity expectations (q 21 >1.00) for this topical activity. In conclusion, none of the activities for which the immunoglobulins can be used as a conventional activity have shown any conventional or locally enhanced mid- to long-term effects on their shared components.
  • This mid- and long-term synergy is probably mainly the secondary synergy of the combined use of the local activity of Ig and the local activities of its shared components, and is also the main mid- and long-term pharmacology of the immunoglobulin composition.
  • Examples 5 and 6 below further investigate reactants that can have a therapeutically meaningful pharmacological response to the above-described local activity of Ig in a local tissue reactor.
  • Nude mice were used as experimental animals and were divided into 4 research series (A, B, C, D) as shown in the table below.
  • Series A, B, C, and D were modeled with the following cancer cells: human pancreas. Cancer Panc-1 cells, human colon cancer Lovo cells, human liver cancer HepG2 cells, human gastric cancer BGC823 cells.
  • the modeling was carried out according to the known method. 1 ⁇ 10 5 cancer cells were subcutaneously injected into the right axilla of each animal, and the modeling was successful when each series grew to the following average volumes: 109 ⁇ 21 mm 3 , 118 ⁇ 18 mm 3 , 106 ⁇ 23mm 3 , 114 ⁇ 27mm 3 .
  • each series that was successfully modeled was randomly divided into 5 groups (groups 0, 1, 2, 3, 4) as shown in the table below, where: group 0 was a negative control group and was given normal saline; group 1 was a negative control group.
  • the chemical ablation control group was given anhydrous ethanol; the group 2 was the common substance group, which was given 2% methylene blue; the group 3 was the immunoglobulin group, which was given 4% human immunoglobulin; the group 4 was the composition group , given 4% human immunoglobulin/2% methylene blue.
  • the medicines are all aqueous solutions, and the research medicines are prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1.
  • group 2 also showed relatively similar short-term efficacy (38.2% ⁇ r7 ⁇ 47.9%) and medium and long-term efficacy (20.3% ⁇ r21 ⁇ 29.2%), in line with the local effect of methylene blue on non-tumor cells Specificity is expected; group 3 did not show short-term efficacy and mid-term and long-term efficacy of therapeutic significance, and its effect was not specific to tumor cells; group 4 also showed relatively similar short-term efficacy (81.3% ⁇ r7 ⁇ 91.4%) and medium and long-term efficacy (70.7% ⁇ r21 ⁇ 80.1%).
  • the tumor cells in this example and other tumor cells in the above examples constitute a tumor cell distribution spectrum with a very broad cellular nature, and the composition of the present invention shows very similar short-term efficacy and medium and long-term efficacy in it, indicating that Ig local
  • the activity has no obvious tumor cell specificity, and can have its pharmacological response with almost any solid tumor tissue, producing similar short-term synergy and mid- and long-term synergy.
  • the local stimulation intensity of each series of group 2 was between groups 0 and 1, and the estimated value was about 2.0; the local stimulation intensity of each series of group 3 was between groups 0 and 2, and the estimated value was about 2.0. is 1.0; the local stimulus intensity of group 4 in each series is between groups 2 and 3, and is estimated to be about 1.2.
  • Example 6 Reactant studies for local activity of Ig: local lesions with fibroblasts
  • Nude mice were used as experimental animals, and they were divided into 2 research series (A, B) as shown in the table below.
  • Series A and B were made of mouse embryonic fibroblasts 3T3 (0.5 ⁇ 10 4 cells/a) and small fibroblasts, respectively.
  • a mixture of murine breast cancer 4T1 cells and mouse embryonic fibroblasts 3T3 (0.5 x 10 5 cells per mouse, 1/10 cell number ratio of 3T3 to 4T1) was used to model pharmacological reactants (nodules).
  • the modeling was carried out according to the known method.
  • the modeled cells were injected subcutaneously in the right axilla of each animal, and the animals of series A and B were selected to grow to 129 ⁇ 32mm3 and 134 ⁇ 27mm3 for the experiment.
  • the mean stromal ratio of the animals in series B was determined to be 39.7%.
  • the successfully modeled series A and B were randomly divided into 5 groups (groups 0, 1, 2, 3, and 4) as shown in the following table, among which: group 0 was a negative control group, given normal saline; group 1 All were chemical ablation control group, given absolute ethanol; group 2 were all shared substance group, given 20% glycine/10% acetic acid (pH3.9); group 3 were all immunoglobulin groups, given 4% human immune globulin Protein; group 4 is a composition group, administered with 4% human immunoglobulin/20% glycine/10% acetic acid (pH 3.9).
  • the medicines are all aqueous solutions, and the test medicines are prepared according to the preparation method of Example 1, and the human immunoglobulins are the freeze-dried intravenous human immunoglobulins in Table 1.
  • the animals in each group of series A and B were given once every 2 days for a total of 3 times.
  • the local irritation intensity (such as struggling intensity) of the experimental animals was observed during administration, and the intensity was estimated by taking the local irritation phenomena of normal saline and absolute ethanol as 1 and 5, respectively.
  • the tumor volume of animals in each series and each group was measured, and the 7-day volume tumor inhibition rate (r7) and The volume tumor inhibition rate (r21) on the 21st day, the results are shown in Table 6 below.
  • the q7 and q21 of the Ig composition calculated by the r7 and r21 of groups 2-4 are both >1.15, indicating that the local activity of Ig provides the same as described above in the tumor-bearing animals of Examples 2-5 Short-term synergy and mid- to long-term synergy beyond expectations of local additive effects.
  • the short-term efficacy (49.6% ⁇ r7 ⁇ 71.2%) and the mid- and long-term efficacy (60.1% ⁇ r21 ⁇ 61.5%) of the Ig composition are not much different. Within the acceptable range of different experiments in the same series (deviation ⁇ 20%).
  • the nodules containing fibroblasts and tumor cells/fibroblasts constitute a broad spectrum of cellular distribution in which the composition of the present invention shows very similar short-term efficacy and mid-term and long-term efficacy, It shows that the local activity of Ig has no obvious cell specificity, and can have its pharmacological reaction with almost any diseased tissue containing fibroblasts, producing similar short-term synergy and mid- and long-term synergy.
  • the local stimulation intensity of each series of group 2 was between groups 0 and 1, and the estimated value was about 3.5; the local stimulation intensity of each series of group 3 was between groups 0 and 2, and the estimated value was about is 1; the local stimulus intensity of group 4 in each series is between groups 2 and 3, estimated to be about 2.5.
  • Example 7 Conditions required for Ig to provide topical activity: topical synergistic studies and optimization
  • the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1 ⁇ 10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model.
  • Successfully modeled experimental animals (with an average tumor volume of 324 mm 3 ) were randomly divided into 1 negative control group (01) and 7 drug study groups (1-7).
  • the negative control is normal saline, and the research drugs are shown in the table below.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1.
  • Each experimental group received intratumoral injection, once every 3 days, for a total of 3 times, and the injection volume was 150 ⁇ l/only/time.
  • the animals were euthanized, the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 7.
  • 5-fluorouracil used in single drug group 3 is an effective tumor cell inhibitory drug for both conventional and intratumoral drugs
  • quinine dihydrochloride used in single drug group 4 is an intratumoral drug concentration ⁇ 3.
  • methylene blue used in single drug group 2 is a conventional vital dye that can only be used as chemical ablative agents when the concentration is greater than or equal to 1.
  • the tumor inhibition rates of single drug groups 2, 3, and 4 were 13%, 53%, and 58%, respectively.
  • the tumor inhibition rate of the composition group 5 was more than 135% of that of the other two groups.
  • composition groups 5, 6, 7 were >1.15, ⁇ 1.15 and ⁇ 1.15, respectively, showing clearly different synergistic effects. Moreover, the significant difference in efficacy among the composition groups 5, 6, and 7 was caused by the presence or absence of obvious synergistic efficacy.
  • the local immunoglobulin activity does not locally synergize with cytotoxic antitumor drugs such as 5-fluorouracil or antimalarial drugs with chemical ablative effects such as quinine dihydrochloride, but can locally synergize in experimental Methylene blue, which does not show significant chemical ablation under conditions.
  • the local synergistic pharmacology of the composition of the present invention does not necessarily require the immunoglobulin to have local efficacy, or even does not necessarily require the immunoglobulin synergist to have effective local efficacy. There is no necessary relationship between immunoglobulin synergist and whether it shows effective activity against tumor cells or whether it shows effective activity for chemical ablation.
  • This local synergistic pharmacology appears to be different from the known conventional pathological synergy (eg, immune activity synergy, anti-tumor cell synergy, chemical ablation synergy, antimalarial synergy, etc.) Synergistic Pharmacology.
  • pathological synergy eg, immune activity synergy, anti-tumor cell synergy, chemical ablation synergy, antimalarial synergy, etc.
  • the immunoglobulin synergist is preferably one or more conventional ineffective drugs selected from anti-tumor or/and anti-systemic pathogens (parasites, bacteria, viruses), more preferably selected from the following One or more of the group: amino acid nutrient, acid ablating agent, vital dye.
  • the following experiments further optimize the immunoglobulin synergists.
  • the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1 ⁇ 10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model.
  • Successfully modeled experimental animals (average tumor volume 319 mm 3 ) were randomly divided into 1 negative control group (0) and 11 drug study groups (1-11).
  • the negative control is normal saline, and the research drugs are shown in the table below.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1.
  • Each group was administered once every 3 days for a total of 3 times. 3 days after the end of the administration, the animals were euthanized, the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group.
  • Table 8 The results are shown in Table 8.
  • composition groups 7, 8, 9, 10, and 11 are all >1.15, all showing obvious synergistic efficacy. Comparing between these composition groups, study groups 7, 9, 10, 11 showed effective efficacy (tumor inhibition rate greater than 40%), while study group 8 did not show effective efficacy (tumor inhibition rate less than 40%) , and showed that the difference in tumor weight between each group and the study group 8 was statistically significant (all p ⁇ 0.05).
  • amino acid nutrients in the immunoglobulin/amino acid nutrient composition of the present invention can be selected from a wide range, preferably amino acids or their salts selected from the group consisting of: Oligopeptides and polypeptides comprising or consisting of the following amino acids: arginine, glycine, cysteine, alanine, serine, glutamic acid.
  • the experimental animals were BALB/c mice, and the model cells were melanoma cells, and 100ul (2.5 ⁇ 10 6 cells per milliliter) per animal were subcutaneously transplanted into the right armpit of the animal to model the tumor.
  • Successfully modeled experimental animals (average tumor volume 238 mm 3 ) were randomly divided into 1 negative control group (0) and 9 drug study groups (1-9).
  • the negative control is normal saline, and the research drugs are shown in the table below.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each group was administered once every 3 days for a total of 3 times.
  • the animals were euthanized 3 days after the drug administration, and the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group.
  • the results are shown in Table 9.
  • composition groups 6, 7, 8, and 9 are all >1.15, all showing obvious synergistic efficacy. Comparing between these composition groups, study groups 6, 7, 8 showed effective efficacy (tumor inhibition rate greater than 40%), while study group 9 did not show effective efficacy (tumor inhibition rate less than 40%), and The difference in tumor weight between each group and study group 9 was statistically significant (all p ⁇ 0.05).
  • the vital dye in the immunoglobulin/vital dye composition of the present invention is preferably selected from methylene blue type vital dyes, more preferably selected from methylene blue and its derivatives.
  • Embodiment 8 Ig provides the conditions required for local activity: Ig pharmacological concentration research and optimization
  • the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1 ⁇ 10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model.
  • the successfully modeled experimental animals (average tumor volume 386mm 3 ) were randomly divided into 1 negative control group (01) and 14 drug study groups (1-14).
  • the negative control is normal saline, and the research drugs are shown in the table below.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1.
  • co-drugs of the same dose and ratio can show substantial differences sharing effect.
  • the actual/expected ratio q of composition groups 5, 9 and 13 were all >1.15, all showing obvious synergistic efficacy.
  • the actual/expected ratio q of composition groups 6, 10, and 14 were all ⁇ 1.15, and none of them showed synergistic efficacy.
  • composition of conventional and topical medications has completely different implications in terms of target pharmacy.
  • a very different drug mixture will be formed in the same target area (eg, tumor).
  • Conventional drugs are distributed in the blood and then reach the target area with the blood.
  • the original mixture no longer exists.
  • Different drug components are present in the new blood-drug mixture in different concentrations (often more than a hundredfold dilution), and even different complexes.
  • the local administration of the drug mixture relies on external force to directly enter the target area, and its composition in the target area is almost the same as its administration composition. It is entirely possible that these two very different compositions of the same drug mixture in the target area have their own targets and thus their own pharmacology.
  • the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1 ⁇ 10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model.
  • Successfully modeled experimental animals (average tumor volume 297mm 3 ) were randomly divided into 1 negative control group (0) and 7 drug study groups (1-7).
  • the negative control is normal saline, and the research drugs are shown in the table below.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1.
  • Each group was administered once every 3 days, a total of 3 times, and intratumoral injection was performed, and the amount of each injection was shown in Table 11 below.
  • composition group 7 further confirms that the occurrence of this threshold is not accidental.
  • the tumor inhibition rate further increased with the increase of the concentration of immunoglobulin, showing a higher synergistic effect.
  • the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 0.5 ⁇ 10 5 cells per animal were subcutaneously transplanted into the right armpit of the animal to model.
  • Successfully modeled experimental animals (average tumor volume 259 ⁇ 32 mm 3 ) were randomly divided into 1 negative control group (0) and 7 drug study groups (1-7).
  • the negative control is normal saline, and the research drugs are shown in the table below.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1.
  • Each group was administered once every 3 days, a total of 3 times, and intratumoral injection was performed, and the amount of each injection was shown in the table below.
  • the tumor volume of animals in each series and each group was measured, and the 7-day volume tumor inhibition rate (r 7 ) and The volume tumor inhibition rate (r 21 ) on the 21st day is shown in Table 12 below.
  • q 7 and q of the human immunoglobulin/methylene blue composition calculated by r 7 and r 21 of groups 1, 2, 5, 1, 3, 6, and 1, 4, 7 21 are all >1.15, indicating that the local activity of high concentrations of Ig provides both short-term synergy and mid- to long-term synergy beyond the expectations of local additive effects as described in Examples 2-4 above.
  • q 21 increased with the increase of Ig concentration, which indicated that the mid- and long-term synergy had a certain Ig concentration dependence.
  • a necessary condition for an immunoglobulin to provide local activity or to act as a local synergist in the compositions of the present invention is that the concentration of the immunoglobulin must be >1.5%, preferably 2.0 %-30% or 3.0%-30%.
  • the immunoglobulin may be used as a conventional immune drug, but cannot be used as a topical active ingredient in the present invention to prepare a topical drug for the treatment of local pathological diseases.
  • Embodiment 9 provides the conditions required for local activity: the study and optimization of the pharmacological concentration of the concomitant
  • the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1 ⁇ 10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model.
  • Successfully modeled experimental animals (average tumor volume 311 mm 3 ) were randomly divided into 1 negative control group (0) and 9 drug study groups (1-9).
  • the negative control is normal saline, and the research drugs are shown in the table below.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1.
  • composition group 9 further confirms that the occurrence of this threshold is not accidental.
  • the tumor inhibition rate further increased with the increase of the concentration of reduced glutathione, showing a higher synergistic effect.
  • concentration thresholds eg, 15%, 20%, etc.
  • a necessary condition for immunoglobulin to provide local activity such that amino acid nutrients act as their local synergists is that the concentration ratio, not the quantitative ratio, of immunoglobulin and amino acid nutrients satisfies the following conditions:
  • the concentration of the immunoglobulin must be the preferred concentration in Example 8 above, and the concentration of the amino acid nutrients must be >7%, ⁇ 8%, preferably 8-30% or 10-30%.
  • the amino acid nutrients may be used as adjuvants (solubilizers, stabilizers, etc.) of immunoglobulin when the local administration concentration is less than the synergistic concentration, but cannot be used as the local synergistic agent of immunoglobulin in the present invention. Preparation of topical medicaments for the treatment of local pathological diseases.
  • the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1 ⁇ 10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model.
  • Successfully modeled experimental animals (with an average tumor volume of 307 mm 3 ) were randomly divided into 1 negative control group (0) and 9 drug study groups (1-9).
  • the negative control is normal saline, and the research drugs are shown in the table below.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1.
  • composition group 8 when the administration concentration of methylene blue in the composition reached a threshold ( ⁇ 0.35%), the difference in tumor weight between composition group 8 and negative control group 01 was statistically significant (p ⁇ 0.05), and its actual The /expected ratio q is >1.15, showing a clear synergistic effect.
  • Composition group 9 further confirms that the occurrence of this threshold is not accidental.
  • the tumor inhibition rate At the same methylene blue dose, the tumor inhibition rate further increased with the increase of methylene blue concentration, showing a higher synergistic effect.
  • the experimental animals were nude mice, the model cells were human lymphoid Raji cells, and 1 ⁇ 10 6 cells per animal were used to model the transplanted tumor subcutaneously in the right armpit of the animal.
  • Successfully modeled experimental animals (average tumor volume 127 ⁇ 19 mm 3 ) were randomly divided into 1 negative control group (0) and 5 drug study groups (1-5).
  • the negative control is normal saline, and the research drugs are shown in the table below.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1.
  • Each group was administered once every 3 days, a total of 3 times, and intratumoral injection was performed, and the amount of each injection was shown in the table below.
  • the tumor volume of animals in each series and each group was measured, and the 7-day volume tumor inhibition rate (r 7 ) and The volume tumor inhibition rate (r 21 ) on the 21st day is shown in Table 15 below.
  • the q7 and q21 of the human immunoglobulin/methylene blue composition calculated by the r7 and r21 of groups 1 , 2 , 4, and 1, 3, 5 are both >1.15, It is demonstrated that the local activity of high concentrations of Ig provides both short-term synergy and mid- to long-term synergy that exceeds the expectations of local additive effects as described in Examples 2-4 above. In addition, q 21 increased with the increase of Ig concentration, indicating that the mid- and long-term synergy had a certain Ig concentration dependence.
  • a necessary condition for immunoglobulin to provide local activity such that vital dye acts as its local synergist is that the concentration ratio of immunoglobulin and vital dye, rather than the amount ratio, satisfies the following condition:
  • the concentration of globulin must be the preferred concentration in Example 8 above, while the concentration of the vital dye must be >0.3%, ⁇ 0.35%, preferably 0.35-10%.
  • the vital dye may be used as an adjuvant of immunoglobulin, but cannot be used as a local synergist of immunoglobulin in the present invention to prepare a topical drug for the treatment of local pathological diseases.
  • Example 10 Conditions required for Ig to provide topical activity: research and optimization of multi-component topical synergists
  • the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1 ⁇ 10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model.
  • Successfully modeled experimental animals (average tumor volume 320mm 3 ) were randomly divided into 1 negative control group (0) and 9 drug study groups (1-9).
  • the negative control is normal saline, and the research drugs are shown in the table below.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1.
  • composition group 8 when the administration concentration of acetic acid in the composition reached a threshold ( ⁇ 2%), the difference in tumor weight between composition group 8 and negative control group 01 was statistically significant (p ⁇ 0.05), and its actual/expected The ratio q is >1.15, showing obvious synergistic effect. Composition group 9 further confirms that the occurrence of this threshold is not accidental. At the same dose of glycine/acetic acid, the tumor inhibition rate further increased with the increase of the concentration of acetic acid, showing a higher synergistic effect.
  • a necessary condition for the amino acid nutrient/acid ablative agent as a topical synergist for immunoglobulins in the compositions of the present invention is that the amino acid nutrient is at a concentration as described in the Examples
  • the acid ablating agent must be a concentration greater than that of its single drug to show chemical ablation efficacy, such as strong acid ⁇ 0.5%, weak acid or weak base (nitrogen-containing weak base, weak acid strong base salt) is ⁇ 2.0% or ⁇ 2.5%.
  • the acid ablating agent is a weak acid
  • its concentration in the composition of the present invention is ⁇ 2.0%, ⁇ 3.0%, preferably 3.0-15% or 5-15%.
  • the synergistic effect of the composition of the present invention can be achieved in that the concentration of the acid ablating agent is much lower than (eg ⁇ 50%, ⁇ 30%, preferably 20%-30%) of its single-agent clinical
  • concentration of the acid ablating agent is much lower than (eg ⁇ 50%, ⁇ 30%, preferably 20%-30%) of its single-agent clinical
  • the same or even higher efficacy is obtained at common concentrations (eg, 50% acetic acid, 7% sodium hydroxide, and 5% hydrochloric acid).
  • the synergistic effect of the composition of the present invention includes both pharmacodynamic synergy and safety synergy.
  • the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1 ⁇ 10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model.
  • Successfully modeled experimental animals (average tumor volume 307 mm 3 ) were randomly divided into 1 negative control group (0) and 10 drug study groups (1-10).
  • the negative control is normal saline, and the research drugs are shown in the table below.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each group was administered once every 3 days for a total of 3 times. Three days after the end of the administration, the animals were euthanized, the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 17.
  • the experimental animals were BALB/c mice, the model cells were breast cancer 4T1 cells, and 100ul (0.5 ⁇ 10 6 cells per milliliter) per animal was used to model the transplanted tumor subcutaneously in the right armpit of the animal.
  • Successfully modeled experimental animals (average tumor volume 237 ⁇ 26 mm 3 ) were randomly divided into 1 negative control group (0) and 7 drug study groups (1-7).
  • the negative control is normal saline
  • the research drugs are shown in the following table
  • the Ig compound is the composition of methylene blue and polysaccharide
  • the model drug of the polysaccharide is carboxymethyl glucan (yeast source) , water-soluble beta-D-glucan (yeast source), and dextran.
  • the medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1.
  • Each group was administered once every 2 days, a total of 2 times, intratumoral injection, each injection volume was 100ul.
  • the tumor volume of animals in each series and each group was measured, and the 7-day volume tumor inhibition rate (r 7 ) of groups 1-4 was calculated relative to the negative control group of each series. and 34-day volume tumor inhibition rate (r 21 ), the results are shown in Table 18 below.
  • the q7 and q21 of the Group 5 composition calculated by the r7 and r21 of the Groups 1 , 2, 5 are ⁇ 1.00 and >1.15, respectively; by the r7 of the Groups 1, 3, 6 Calculated q7 and q21 for Group 6 compositions were ⁇ 1.00 and >1.15, respectively; q7 and q21 for Group 7 compositions calculated from r7 and r21 for Groups 1, 4 , 7 q 21 were ⁇ 1.00 and >1.15, respectively, indicating that Ig local activity provided mid- and long-term synergy beyond that expected for mid- and long-term additive effects under conditions that failed to provide short-term synergy.
  • the composition comprises immunoglobulin and two or more multiple immunoglobulin synergists.

Abstract

The present application relates to a new use for an immunoglobulin, a pharmaceutical composition comprising the immunoglobulin and a synergist thereof, and a method for treating topical lesion diseases by means of using the pharmaceutical composition.

Description

包含免疫球蛋白的药物组合物及其应用Pharmaceutical composition comprising immunoglobulin and use thereof 技术领域technical field
本申请公开涉及免疫球蛋白的一种新应用、包含适于该应用的免疫球蛋白及其协同物的局部药物组合物、以及使用该药物组合物治疗局部病变疾病的方法。The present application discloses a novel application of immunoglobulin, a topical pharmaceutical composition comprising an immunoglobulin suitable for the application and a synergist thereof, and a method of using the pharmaceutical composition to treat local pathological diseases.
背景技术Background technique
由于有大量研究工作支持，实体肿瘤常被用作局部病变疾病、尤其是难治性局部病变疾病的研究模型。实体肿瘤是一种具有瘤体征状的肿瘤疾病，瘤体是包含有肿瘤细胞的一种特征性病变组织。以胰腺癌瘤体为例，胰腺癌细胞在其中仅占约30％的体积比。可见，除了肿瘤细胞，瘤体组织中往往还存在更大量的其它组成(有时亦被称作肿瘤细胞的微环境)，包括其它多种细胞(例如成纤维细胞)、多种细胞间质(例如纤维)、多种管道等。Solid tumors are often used as models for the study of localized disease, especially refractory localized disease, supported by a large amount of research work. Solid tumor is a tumor disease with tumor symptoms, and the tumor body is a characteristic diseased tissue containing tumor cells. Taking pancreatic cancer tumor as an example, pancreatic cancer cells only account for about 30% of the volume. It can be seen that in addition to tumor cells, there are often a larger number of other components (sometimes also referred to as tumor cell microenvironment) in tumor tissue, including other various cells (such as fibroblasts), various intercellular substances (such as fiber), various pipes, etc.
局部给药具有将药物进行物理靶向的优点。于是曾经有人认为，细胞毒药物的局部给药可以提高其瘤内浓度、从而提高其疗效。然而，局部给药细胞毒药物却并未显示出疗效的大不相同的提高。仅仅提高其瘤内浓度，似乎尚不能大大提高其靶向瘤内组织中癌细胞的效率,反而明显地增加了局部刺激性。于是,除了求助于其缓释形式以外，细胞毒药物在临床上几乎仍全身给药。临床上用过的局部活性药物例如化学消融剂(高纯度乙醇、高浓度酸碱、弱酸强碱盐)不以细胞破坏而以组织破坏为特征。与细胞毒药物相比，其几乎无靶向癌细胞的全身疗效，却往往显示出更高局部疗效。然而，它们通常是不能对靶组织和其它组织作足够区别的強破坏剂。这使得它们实际能够应用的介入体积(例如酸碱用量不超过0.2ml/kg)和介入部位非常受限(例如对瘤体所在器官的限制、瘤体边缘消融受限等)。此外,它们通常并无中长期作用极易反弹。因而，近十年来化学消融剂以经从恶性实体肿瘤临床上淡出。实际上，目前临床上以几乎没有局部安全性和短期局部疗效均高的局部药物,更没有中长期疗效的局部药物。Topical administration has the advantage of physically targeting the drug. Therefore, it was once believed that local administration of cytotoxic drugs can increase their intratumoral concentration, thereby improving their efficacy. However, topical administration of cytotoxic drugs did not show a disparate improvement in efficacy. Merely increasing its intratumoral concentration does not seem to greatly improve the efficiency of targeting cancer cells in intratumoral tissues, but significantly increases local irritation. Thus, cytotoxic drugs are still clinically administered almost systemically, except for recourse to their sustained-release forms. Clinically used topically active drugs such as chemical ablatives (high-purity ethanol, high-concentration acid-base, weak-acid-strong-base salt) are not characterized by cell destruction but tissue destruction. Compared with cytotoxic drugs, they have almost no systemic efficacy targeting cancer cells, but often show higher local efficacy. However, they are generally strong disruptors that do not sufficiently distinguish the target tissue from other tissues. This makes them practically applicable to the intervention volume (for example, the amount of acid and alkali does not exceed 0.2ml/kg) and the intervention site is very limited (for example, the limitation of the organ where the tumor is located, the ablation of the tumor edge, etc.). In addition, they usually have no mid- to long-term effect and are easily rebound. As a result, chemical ablative agents have been clinically faded from malignant solid tumors in the past decade. In fact, there are almost no local drugs with high local safety and short-term local efficacy in clinical practice, and there is no local drug with medium and long-term efficacy.
因而，仍然需要开发新的治疗诸如实体肿瘤的局部病变疾病的药物、尤其是局部药物、更尤其是有中长期疗效的局部药物，以满足现有技术尚不能满足的各种临床需求。实际上，其它局部病变、尤其是顽固性局部病变疾病的防治也都有这个急迫需要。Therefore, there is still a need to develop new drugs for the treatment of local diseased diseases such as solid tumors, especially topical drugs, more particularly topical drugs with mid- and long-term efficacy, to meet various clinical needs that cannot be met by the existing technologies. In fact, the prevention and treatment of other local lesions, especially intractable local lesions also have this urgent need.
发明内容SUMMARY OF THE INVENTION
有鉴于此，本发明的目的在于提供一种预防和治疗局部病变疾病、尤其是难治性局部病变疾病的更高综合性能(例如局部安全性、局部短期疗效、或/和中长期疗效)的局部药物。更具体而言，本发明的目的在于提供一种基于免疫球蛋白(下文简称为Ig)的一种新发现的局部活性-即，使得一个或多个常规无效药物转化为其短期协同作用、局部协同安全性、或/和中长期协同作用共用物，从而制备的局部药物组合物(下文简称为药物组合物)，实现高效低毒，尤其是中长期内，以为临床提供各种选择。In view of this, the purpose of the present invention is to provide a higher comprehensive performance (for example, local safety, local short-term efficacy, or/and medium and long-term efficacy) for the prevention and treatment of local diseased diseases, especially refractory local diseased diseases. topical medication. More specifically, the object of the present invention is to provide a newly discovered topical activity based on immunoglobulins (hereinafter referred to as Ig) - that is, to convert one or more conventional ineffective drugs into their short-term synergistic, topical Synergistic safety, or/and mid- and long-term synergistic effects, the prepared topical pharmaceutical composition (hereinafter referred to as pharmaceutical composition) achieves high efficiency and low toxicity, especially in the medium and long term, providing various options for clinical use.
根据本申请公开的一个方面，其提供免疫球蛋白作为可提供局部协同作用的局部活性成分在制备可致靶区组织坏死的药物组合物中的应用,其中所述药物组合物还包含所述免疫球蛋白的局部协同物、以及药物学可接受的合适载体。According to one aspect disclosed in the present application, it provides the use of immunoglobulin as a local active ingredient that can provide local synergy in the preparation of a pharmaceutical composition that can cause tissue necrosis in a target area, wherein the pharmaceutical composition further comprises the immune A topical synergist for globulin, and a suitable pharmaceutically acceptable carrier.
根据本申请公开的另一个方面，其提供一种药物组合物，其包含可提供局部协同作用的免疫球蛋白、所述免疫球蛋白的局部协同物、以及药物学可接受的合适载体。According to another aspect disclosed in the present application, there is provided a pharmaceutical composition comprising an immunoglobulin that can provide a local synergistic effect, a local synergist of the immunoglobulin, and a suitable pharmaceutically acceptable carrier.
根据本申请公开的再一个方面，其提供一种治疗或缓解局部病变疾病的方法，其包括可致靶区组织坏死的药物组合物的施用,其中所述药物组合物包含可提供局部协同作用的免疫球蛋白、所述免疫球蛋白的局部协同物、以及药物学可接受的合适载体。According to yet another aspect disclosed in the present application, there is provided a method of treating or ameliorating local pathological diseases, comprising the administration of a pharmaceutical composition that can cause tissue necrosis in a target area, wherein the pharmaceutical composition comprises a local synergistic effect. Immunoglobulins, topical synergists of the immunoglobulins, and suitable pharmaceutically acceptable carriers.
在一个实施方案中，所述局部协同作用所需条件包括:所述免疫球蛋白以浓度(w/v)>1.5％、≥2.0、2.0-35％，优选3.0-30％进入所述靶区。In one embodiment, the conditions required for the local synergy include: the immunoglobulin enters the target region at a concentration (w/v) >1.5%, >=2.0, 2.0-35%, preferably 3.0-30% .
根据本发明的实施方案与现有技术中包含免疫球蛋白的药物组合物相比具有以下优点：可提供靶区组织有效坏死、甚至消融作用,同时还有可能保留免疫球蛋白的免疫(调节)作用以及提供更高的局部安全性和/或更好的中长期疗效。Embodiments according to the present invention have the following advantages over prior art pharmaceutical compositions comprising immunoglobulins: effective necrosis, or even ablation, of target tissue can be provided, while also potentially preserving the immunity (modulation) of immunoglobulins effect and provide higher local safety and/or better mid- and long-term efficacy.
根据本发明的实施方案与局部病变疾病治疗的其它现有技术相比具有以下优点：与现有分子靶向药物相比，显示出不那么苛刻的适应症筛选(可靶向局部病变中的成纤维细胞或/和泛实体肿瘤细胞以及与它们相关的细胞间质)，以及例如针对快速生长瘤体、大瘤体和乏血供瘤体的巨大潜力；与现有局部活性药物例如化学消融剂相比，显示出更高有效性(尤其是常规化学消融剂所无的中长效性)和明显低得多的局部刺激性以及对周边正常组织的伤害，从而可以有较大的介入适应范围和较高的应用体积。本发明的应用和组合物也不受现有细胞毒性药物和现有分子靶向药物遭遇的耐药性问题的困扰。此外，该应用和组合物制备方便、成本便宜，特别有助于使难以承受高额费用的广大人群也享受到安全、有效治疗。Embodiments according to the present invention have the following advantages over other prior art for focal disease treatment: show less stringent screening of indications (can target pathogens in focal disease) compared to existing molecularly targeted drugs fibroblasts or/and pan-solid tumor cells and their associated interstitial cells), and great potential, for example, against rapidly growing tumors, large tumors, and hypovascular tumors; with existing locally active drugs such as chemical ablative agents Compared with conventional chemical ablative agents, it shows higher efficacy (especially the medium and long-term effect that conventional chemical ablation agents do not have) and significantly lower local irritation and damage to surrounding normal tissues, so that a wider range of interventional adaptations can be achieved. and higher application volumes. The applications and compositions of the present invention are also not plagued by drug resistance problems encountered with existing cytotoxic drugs and existing molecularly targeted drugs. In addition, the application and composition are convenient to prepare and low cost, and are particularly helpful for making safe and effective treatment available to a large number of people who cannot afford high costs.
具体实施方式Detailed ways
本发明的发明人在一个通常用作化疗模型的荷瘤裸鼠实验中意外地发现，免疫球蛋白(Ig)虽不能单独提供局部作用，但可以与一些优选物质在某些特定的条件下，产生局部协同作用从而有效治疗、甚至是消融肿瘤瘤体组织。进一步的研究还显示可产生中长期协同作用从而减弱局部药物通常具有的中长期反弹、或/和局部协同安全性。这些特定条件并非Ig常规组合物中的组成，而是如以下所限定的。The inventors of the present invention unexpectedly found in a tumor-bearing nude mouse experiment that is usually used as a chemotherapy model, although immunoglobulin (Ig) alone cannot provide local effects, but can be combined with some preferred substances under certain specific conditions, Local synergy is produced to effectively treat and even ablate tumor tissue. Further studies have also shown that mid- to long-term synergy can be produced to reduce the mid- to long-term rebound, or/and local synergistic safety that topical drugs typically have. These specific conditions are not constituents in conventional compositions of Ig, but are as defined below.
在本申请公开的一个方面中，其提供免疫球蛋白作为可提供局部协同作用的协同物在制备可致靶区组织坏死的药物组合物中的应用,其中所述药物组合物还包含所述免疫球蛋白的局部协同物、以及药物学可接受的合适载体。In one aspect disclosed in the present application, it provides the use of immunoglobulin as a synergist that can provide local synergy in the preparation of a pharmaceutical composition that can cause tissue necrosis in a target area, wherein the pharmaceutical composition further comprises the immune A topical synergist for globulin, and a suitable pharmaceutically acceptable carrier.
在本申请公开的另一个方面中，其提供一种药物组合物，其包含可提供局部协同作用的免疫球蛋白、所述免疫球蛋白的局部协同物、以及药物学可接受的合适载体。In another aspect disclosed herein, it provides a pharmaceutical composition comprising an immunoglobulin that can provide local synergy, a local synergist of the immunoglobulin, and a suitable pharmaceutically acceptable carrier.
在本申请公开的又一个方面中，其提供一种治疗或缓解局部病变疾病的方法，其包括可致靶区组织坏死的药物组合物的施用,其中所述药物组合物包含可提供局部协同作用的免疫球蛋白、所述免疫球蛋白的局部协同物、以及药物学可接受的合适载体。In yet another aspect disclosed in the present application, there is provided a method of treating or ameliorating a local pathological disease, comprising the administration of a pharmaceutical composition that can cause tissue necrosis in a target area, wherein the pharmaceutical composition comprises a composition that can provide a local synergistic effect of immunoglobulins, topical synergists of said immunoglobulins, and suitable pharmaceutically acceptable carriers.
在本申请公开的又一个方面中，其提供一种用于治疗局部病变疾病的装置，其包含根据本申请公开所定义的包含免疫球蛋白的局部药物组合物。In yet another aspect of the present disclosure, there is provided a device for the treatment of a localized disease comprising a topical pharmaceutical composition comprising immunoglobulin as defined in the present disclosure.
在本申请公开的范围中，所用术语“免疫球蛋白”是一个超越其在现有技术中的常规功能的结构概念，是指所有与本申请实施例中所用免疫球蛋白在组成上相同或相似、且可提供本申请公开的局部活性的多肽,而这些多肽在现有技术中仅发现并应用了它的免疫(调节)作用。免疫球蛋白包括天然免疫球蛋白及其工程类似物。其中所述天然免疫球蛋白包括免疫球蛋白G(IgG)、免疫球蛋白A(IgA)、免疫球蛋白M(IgM)、免疫球蛋白D(IgD)、免疫球蛋白E(IgE)，通常从动物血漿的分离获得，例如静注免疫球蛋白G(IgG)、静注免疫球蛋白A(IgA)、静注免疫球蛋白M(IgM)等等。其中所述天然免疫球蛋白的工程类似物是指不同于天然免疫球蛋白、但具有天然免疫球蛋白的全部或部分关健结构的工程制品(包括基因工程制品)，例如免疫球蛋白复合物,以下免疫球蛋白片断及其复合物(免疫球蛋白Fc片断、免疫球蛋白Fab片断、免疫球蛋白Fab’片断、免疫球蛋白F(ab’)片断、单健免疫球蛋白),等等。In the scope of the present disclosure, the term "immunoglobulin" is used as a structural concept beyond its conventional function in the prior art, and refers to all immunoglobulins that are identical or similar in composition to the immunoglobulins used in the examples of the present application , and can provide the locally active polypeptides disclosed in this application, and these polypeptides have only been found and used in the prior art for their immune (modulating) effects. Immunoglobulins include natural immunoglobulins and their engineered analogs. Wherein the natural immunoglobulin includes immunoglobulin G (IgG), immunoglobulin A (IgA), immunoglobulin M (IgM), immunoglobulin D (IgD), immunoglobulin E (IgE), usually from Obtained by separation of animal plasma, such as intravenous immunoglobulin G (IgG), intravenous immunoglobulin A (IgA), intravenous immunoglobulin M (IgM) and the like. Wherein the said engineered analogs of natural immunoglobulins refer to engineered products (including genetically engineered products) that are different from natural immunoglobulins but have all or part of the key structures of natural immunoglobulins, such as immunoglobulin complexes, The following immunoglobulin fragments and their complexes (immunoglobulin Fc fragment, immunoglobulin Fab fragment, immunoglobulin Fab' fragment, immunoglobulin F (ab') fragment, single-linked immunoglobulin), and the like.
在本申请公开的范围中，所用术语“靶区”是指给药的目标部位，例如局部病变的相邻处、界面、内部(优选为内部)等等。所用术语“靶区组织”是指给药的目标组织,即局部病变组织,例如包含癌细胞或/和成纤维细胞的局部病变组织。例如，当病变为肿瘤时，靶为肿瘤，靶区为瘤区(例如瘤体内或/和药物可进入瘤体内的周边)；当病变为非瘤肿大时，靶为肿块，靶区为肿大区(例如增生、囊肿、结节等病变处或/和可进入其中的周边)；当病变为局部炎症时，靶区为发炎区(例如发炎肿块或/和药物可进入肿块的周边)；当病变为分泌异常组织时，靶为分泌异常的分泌腺，靶区则为包含这些异常分泌腺的组织或器官或/及其周边。In the context of the present disclosure, the term "target volume" is used to refer to the target site of administration, eg, adjacent to a localized lesion, interface, interior (preferably interior), and the like. The term "target tissue" as used refers to the target tissue for administration, ie, a locally diseased tissue, eg, a locally diseased tissue containing cancer cells or/and fibroblasts. For example, when the lesion is a tumor, the target is the tumor, and the target area is the tumor area (for example, within the tumor or/and the periphery of the tumor where the drug can enter); when the lesion is non-tumor, the target is the tumor, and the target area is the tumor Large areas (such as hyperplasia, cysts, nodules, etc., or/and the surrounding area that can enter); when the lesion is localized inflammation, the target area is the inflamed area (such as an inflamed mass or/and the surrounding area where drugs can enter); When the lesions are abnormal secretory tissues, the target is the secretory glands with abnormal secretion, and the target area is the tissues or organs containing these abnormal secretory glands or/and their surroundings.
在一个实施方案中，所述药物组合物优选为液体剂型,所述载体包括水。In one embodiment, the pharmaceutical composition is preferably in a liquid dosage form, and the carrier includes water.
在一个实施方案中，所述免疫球蛋白包括多克隆抗体、优选包括人免疫球蛋白及其工程类似物、更优选包括人IgG及其工程类似物。在一个实施方案中，所述免疫球蛋白为人IgG(包括静注免疫球蛋白、肌注免疫球蛋白)或其工程类似物(包括基因工程IgG)。在一个实施方案中，所述免疫球蛋白选自现有技术中的抗肿瘤单克隆抗体之外的免疫球蛋白,其中所述抗肿瘤单克隆抗体是指在现有技术中被单独使用、与常规化疗药物共用、与放疗共用可有效***的抗肿瘤特异性免疫球蛋白。In one embodiment, the immunoglobulins include polyclonal antibodies, preferably human immunoglobulins and engineered analogs thereof, more preferably human IgG and engineered analogs thereof. In one embodiment, the immunoglobulin is human IgG (including intravenous immunoglobulin, intramuscular immunoglobulin) or an engineered analog thereof (including genetically engineered IgG). In one embodiment, the immunoglobulin is selected from immunoglobulins other than anti-tumor monoclonal antibodies in the prior art, wherein the anti-tumor monoclonal antibodies refer to those used alone in the prior art, which are combined with Anti-tumor-specific immunoglobulins that can effectively treat tumors are shared with conventional chemotherapeutic drugs and with radiotherapy.
在本申请公开的药物组合物中，所述免疫球蛋白所提供的局部活性包括通过与其局部协同物在某些特定条件下的共用形成以下局部协同作用之至少一种：短期局部协同作用、中长期局部协同作用、局部刺激性拮抗。在本申请公开中,所述免疫球蛋白及其局部协同物 共用的特定条件包括特定的共用反应器(所述靶区组织)和进入共用反应器的特定浓度(见以下免疫球蛋白浓度以及与其局部协同物的共用量比如重量比)。在本申请公开中，免疫球蛋白的局部协同物有时又简称为免疫球蛋白的共用物。本申请公开的药物组合物的药理作用包括在所述靶区组织内的局部协同作用(例如局部共用产生的短期协同作用或/和中长期协同作用)以及任选存在的其它药理作用。In the pharmaceutical composition disclosed in the present application, the local activity provided by the immunoglobulin includes forming at least one of the following local synergistic effects through the sharing of its local synergist under certain specific conditions: short-term local synergy, medium Long-term local synergy, local irritant antagonism. In the disclosure of the present application, the specific conditions shared by the immunoglobulin and its local synergist include a specific common reactor (the target tissue) and a specific concentration entering the common reactor (see the following immunoglobulin concentrations and its The shared amount of topical synergist such as weight ratio). In the present disclosure, local synergists of immunoglobulins are sometimes referred to simply as immunoglobulin synergists. The pharmacological effects of the pharmaceutical compositions disclosed in the present application include local synergistic effects (eg, short-term synergy or/and mid- and long-term synergies produced by topical co-administration) and other optional pharmacological effects in the target tissue.
在一个实施方案中，所述Ig及其共用物的协同作用并非是它们的常规抗病原体活性(例如细胞毒、病毒毒或细菌毒活性)的联合作用所能提供的协同作用、或它们的任何常规活性(例如免疫调节作用与细胞毒)的联合作用所能提供的协同作用,而是主要由它们的局部活性的联合作用所能提供的局部协同作用。在一个实施方案中，所述协同作用包括局部化学协同作用(显示为短期协同疗效)和任选存在的其它药理作用。在一个实施方案中，所述协同作用包括中长期协同作用(显示为中长期协同疗效)和任选存在的其它药理作用。在一个实施方案中，所述中长期协同作用包括所述局部化学协同作用的次生免疫协同作用和任选存在的其它免疫作用。In one embodiment, the synergy of the Ig and its conjugates is not that provided by the combination of their conventional antipathogenic activities (eg, cytotoxic, viral or bacterial activity), or any of their The synergy provided by the combination of conventional activities (eg, immunomodulatory and cytotoxic) is primarily the local synergy provided by the combination of their local activities. In one embodiment, the synergistic effect includes local chemical synergy (shown as a short-term synergistic effect) and optionally other pharmacological effects. In one embodiment, the synergistic effect includes mid- to long-term synergy (shown as mid- to long-term synergistic efficacy) and optionally other pharmacological effects. In one embodiment, the mid- to long-term synergy includes secondary immune synergy of the local chemical synergy and optionally other immune effects.
在本申请公开的范围中，所述药物组合物为局部给药剂型。Within the scope of the present disclosure, the pharmaceutical composition is in a topical dosage form.
在本发明的范围内，“局部给药剂型”区别于常规给药剂型,后者是指基于常规活性、且药物组成也适合于全身性给药、从而以全身性给药为主的剂型；前者则是指基于局部活性、且药物组成也适合于该靶区环境的要求、从而只能依靠介入方式直接递送至靶区的药物剂型。不同剂型通常会导致不同的药物组成限制,例如局部给药剂型通常须排除常规剂型中通常包含的pH调节剂(使药物尽可能中性)、渗透压调剂(使药物尽可能靠近生理渗透压)、稀释液(通常使用pH和渗透压尽可能靠近生理条件者)选择等,因为这些常规剂型中通常包含的辅料(例如NaCl)有可能干扰局部给药剂型中的局部活性。Within the scope of the present invention, "topical dosage forms" are distinguished from conventional dosage forms, which refer to dosage forms based on conventional activities and whose pharmaceutical composition is also suitable for systemic administration, so that systemic administration is predominant; The former refers to a pharmaceutical dosage form that is based on local activity and whose drug composition is also suitable for the environment of the target area, so that it can only be delivered directly to the target area by interventional means. Different dosage forms usually lead to different drug composition constraints, for example, topical dosage forms usually have to exclude pH adjusters (to make the drug as neutral as possible), osmotic pressure adjusters (to make the drug as close to the physiological osmotic pressure as possible), which are usually included in conventional dosage forms. , diluent (usually using pH and osmotic pressure as close to physiological conditions as possible), etc., because the excipients (such as NaCl) usually included in these conventional dosage forms may interfere with the local activity in the topical dosage form.
在一个实施方案中，所述药物组合物为局部病变内给药剂型。In one embodiment, the pharmaceutical composition is a dosage form for local intralesional administration.
在本申请公开的范围中，术语“局部活性”区别于“常规活性”,后者是指通过常规用药(例如静脉注射、腔内注射、皮下给药、粘膜给药、等等)可获得的活性(例如细胞毒、免疫作用、等等),而前者是指通过常规用药不可获得、只有通过局部给药尤其是靶区给药才能够获得的活性,局部活性包括例如单一药物的局部作用和/或共用药物的局部共用作用。常规活性可通过常规用药后药物经血液吸收形成带药血液后刺激免疫***或进入靶区产生常规药理作用(或常规共用作用),局部活性只能通过局部靶区给药后药物优先在靶区产生药理作用(或共用作用)、包括优先针对靶区组织的化学作用(或共用化学作用)。术语“局部化学作用(或局部共用化学作用)”是指在药物优先在局部组织内而非该组织中的病原体上或身体免疫***内发生的化学作用(或共用化学作用),其显示为短期药效。所用术语“局部活性成分”是指单用或共用时可提供包括有效局部活性在内的药学活性的药物成分。In the context of the present disclosure, the term "topical activity" is distinguished from "conventional activity", which refers to those obtainable by conventional administration (eg, intravenous injection, intracavity injection, subcutaneous administration, mucosal administration, etc.). Activity (such as cytotoxicity, immunity, etc.), while the former refers to the activity that cannot be obtained by conventional medicine, but can only be obtained by local administration, especially target area administration. Local activity includes, for example, the local action of a single drug and /or local co-occurrence of shared drugs. Conventional activity can be achieved through the absorption of the drug through the blood after conventional medication to form drug-carrying blood, and then stimulate the immune system or enter the target area to produce conventional pharmacological effects (or conventional combined effects). Local activity can only be achieved through local target area administration. Pharmacological effects (or co-actions) are produced, including chemical effects (or co-chemical effects) that are preferentially directed against the target tissue. The term "local chemistry (or local shared chemistry)" refers to a chemical action (or shared chemistry) that occurs on a drug preferentially within a local tissue rather than a pathogen in that tissue or within the body's immune system, which appears to be short-term Efficacy. As used, the term "topical active ingredient" refers to a pharmaceutical ingredient which, when used alone or in combination, provides pharmaceutically activity including effective topical activity.
在本申请公开的范围中，术语“共用作用”是指多个(两个或两个以上)活性组分被混合后施用至体内产生的共用药理(或药效)作用或共用毒理(或副)作用,其通常具有高度不确定性。若共用作用为各组分单用作用之和,称之为“相加作用”；若共用作用不及各组分单用作用 之和,称之为“拮抗作用”：若共用作用超过各组分单用作用之和,称之为“协同作用”,例如产生本发明实施例中约定的实际/预期比q>1.15的药效的作用。术语“局部协同作用”是指局部共用、尤其是在局部病变内的共用所形成的协同作用,包括基于短期药效的短期协同作用和基于中长期药效的中长期协同作用。术语“局部协同物”是指通过局部共用可产生局部协同作用的药物组分。在本申请公开的范围内，术语“治疗有效量”是指用于治疗疾病(例如肿瘤)并获得有效效果(例如降低或/和缓解疾病症状)的药物的剂量。In the context of the present disclosure, the term "co-action" refers to the co-pharmacological (or pharmacodynamic) action or co-toxicological (or co-toxicological (or side effects), which are often highly uncertain. If the joint action is the sum of the individual actions of each component, it is called "additive action"; if the joint action is less than the sum of the individual actions of each component, it is called "antagonism": if the joint action exceeds the sum of the individual components The sum of the single effects is called "synergistic effect", for example, the effect of producing the actual/expected ratio q>1.15 of the medicinal effect agreed in the examples of the present invention. The term "local synergy" refers to the synergy formed by local use, especially in local lesions, including short-term synergy based on short-term efficacy and mid- and long-term synergy based on mid- and long-term efficacy. The term "topical synergist" refers to a pharmaceutical component that produces a local synergistic effect by topical administration. Within the scope of this disclosure, the term "therapeutically effective amount" refers to a dose of a drug that is used to treat a disease (eg, tumor) and achieve an effective effect (eg, reduce or/and alleviate symptoms of the disease).
本申请公开的药物组合物如此制备、组成或施用,使其满足所述Ig能提供所述局部活性、或所述局部协同作用所需的条件,从而通过致靶区组织坏死、优选有效坏死、更优选为中长期反弹最小化的坏死来治疗局部病变疾病。在一个实施方案中，所述组织坏死包括组织直接坏死和任选存在的组织间接坏死。在一个实施方案中，所述组织直接坏死如此之强烈以至于组织化学消融。The pharmaceutical compositions disclosed herein are prepared, composed or administered such that the conditions required for the Ig to provide the local activity, or the local synergistic effect, are achieved by causing tissue necrosis, preferably effective necrosis, in the target area. More preferably, necrosis with minimal rebound in the medium to long term is used to treat localized disease. In one embodiment, the tissue necrosis includes direct tissue necrosis and optionally indirect tissue necrosis. In one embodiment, the direct necrosis of the tissue is so severe that it is histochemically ablated.
在本申请公开的范围中，所用术语“组织间接坏死”是指通过对该靶区组织中的病原体(例如细胞、病毒或细菌)增殖抑制或对个体免疫***刺激所产生的效应所导致的组织坏死,前者例如细胞毒药物通过抑制癌细胞有可能最终导致癌细胞所在的组织坏死,后者例如抗肿瘤疫苗通过激发免疫***针对癌细胞的攻击有可能最终导致癌细胞所在的组织坏死,该间接坏死于是既可以通过全身性给药也可以通过局部给药来进行。所用术语“组织直接坏死”是指组织破坏因素直接施用于靶区所导致的不依赖于组织间接坏死的组织坏死,其中所述组织破坏因素包括例如:物理消融仪器的消融部件(例如升温针)、常规化学消融剂、等等。In the context of the present disclosure, the term "indirect necrosis of tissue" is used to refer to the effect of the inhibition of the proliferation of pathogens (eg cells, viruses or bacteria) in the target tissue or the stimulation of the immune system of the individual to the tissue Necrosis, the former, such as cytotoxic drugs, may eventually lead to the necrosis of the tissue where the cancer cells are located by inhibiting cancer cells, and the latter, such as anti-tumor vaccines, by stimulating the immune system to attack the cancer cells, may eventually lead to the necrosis of the tissue where the cancer cells are located. Necrosis can then be achieved either by systemic or local administration. The term "direct tissue necrosis" as used refers to tissue necrosis that is independent of indirect tissue necrosis caused by the direct application of a tissue disrupting factor to the target volume, wherein the tissue disrupting factor includes, for example: ablation components of a physical ablation instrument (e.g., warming needles) , conventional chemical ablative agents, etc.
在本发明的范围中，术语“消融”是指原位处理所致局部作用如此之强,以至于靶区组织产生基本上丧失其原有生理功能(失活)的组织坏死。组织坏死包括凝固性坏死、液化性坏死、纤维素样坏死、等等。肿瘤组织坏死可致肿瘤组织形态变化(体积减小),最终显示为抑瘤率升高。术语“化学消融”是指主要是化学性的而非免疫性的消融。术语“化学消融剂”是指局部用药时才能显示出消融效应的药物。In the context of the present invention, the term "ablation" means that the local effect of the in situ treatment is so strong that the target tissue produces tissue necrosis that essentially loses its original physiological function (inactivation). Tissue necrosis includes coagulation necrosis, liquefaction necrosis, fibrinoid necrosis, and the like. Tumor tissue necrosis can cause morphological changes (volume reduction) of tumor tissue, which ultimately manifests as an increase in tumor inhibition rate. The term "chemical ablation" refers to ablation that is primarily chemical rather than immunological. The term "chemical ablative agent" refers to a drug that exhibits an ablative effect only when administered topically.
在本申请公开的范围中，除非另有说明，术语“浓度”是指所述局部药物组合物中指定组分的重量-体积百分比浓度(w/v)。术语“局部给药浓度”是指指定组分在药物实施局部给药时的浓度，其可以是指定组分在药物接触靶区处(例如注射针孔或灌注管出口)的浓度。Within the scope of the present disclosure, unless otherwise stated, the term "concentration" refers to the weight-volume percent concentration (w/v) of the specified component in the topical pharmaceutical composition. The term "local administration concentration" refers to the concentration of a given component at the time of local administration of the drug, which may be the concentration of the given component at the target area of drug contact (eg, injection needle hole or perfusion tube outlet).
在一个实施方案中，所述药物组合物如此制备、组成或施用以满足所述免疫球蛋白提供所述局部活性所需的以下条件：所述免疫球蛋白以浓度(w/v)为≥2.0、2.0-35％，优选3.0-30％，例如，4％至30％、4-20％、3％-10％、3％-8％、4％-10％，进入所述靶区。In one embodiment, the pharmaceutical composition is prepared, composed or administered such that the immunoglobulin provides the local activity required by the immunoglobulin at a concentration (w/v) of ≥ 2.0 , 2.0-35%, preferably 3.0-30%, eg, 4% to 30%, 4-20%, 3%-10%, 3%-8%, 4%-10%, into the target area.
在本申请公开的范围中，除非另有说明，术语“以某浓度(或量比)进入所述靶区”中的浓度(或量比)(或进入所述靶区的浓度或量比)是指所述活性组分开始接触到所述靶区的浓度(或量比),该浓度(或量比)在本发明的局部(靶区)给药中等同于给药浓度(或给药量比),而在常规(全身性)给药中不等同于给药浓度(或给药量比)。量比，例如重量比。In the scope of the disclosure of the present application, unless otherwise specified, the concentration (or amount ratio) in the term "entering the target area at a certain concentration (or amount ratio)" (or the concentration or amount ratio entering the target area) It refers to the concentration (or amount ratio) at which the active ingredient begins to contact the target area, and this concentration (or amount ratio) is equivalent to the administration concentration (or administration concentration) in the local (target area) administration of the present invention. dose ratio), which is not equivalent to the dosed concentration (or dose ratio) in conventional (systemic) administration. Quantitative ratios, such as weight ratios.
在一个实施方案中，所述免疫球蛋白提供所述局部活性所需的条件还包括:所述局部协 同物选自针对所述靶区的常规无效药物之一种或多种,且所述免疫球蛋白与所述局部协同物以以下量比(重量比)(W 局部协同物/W 免疫球蛋白)进入所述靶区:W 局部协同物/W 免疫球蛋白为≥(0.5-39)/2.0、(0.5-39)/(2.0-35)、或(1-39)/(2.0-35)，优选为(3-30)/2.0，更优选为(4-30)/(2.0-35)或(5-20)/(4.0-30)。 In one embodiment, the conditions required for the immunoglobulin to provide the local activity further include: the local synergist is selected from one or more of conventional ineffective drugs for the target region, and the immune The globulin and the local synergist enter the target area with the following amount ratio (weight ratio) (W local synergist /W immunoglobulin ): W local synergist /W immunoglobulin is ≥ (0.5-39)/ 2.0, (0.5-39)/(2.0-35), or (1-39)/(2.0-35), preferably (3-30)/2.0, more preferably (4-30)/(2.0-35 ) or (5-20)/(4.0-30).
在一个实施方案中，所述免疫球蛋白局部协同物包括或选自针对所述局部病变疾病的常规无效药物之一种或多种。In one embodiment, the immunoglobulin topical synergist comprises or is selected from one or more of conventional ineffective drugs for the localized disease.
在一个实施方案中，所述常规无效药物包括或选自抗肿瘤的常规无效药物。In one embodiment, the conventional ineffective drugs include or are selected from anti-tumor conventional ineffective drugs.
在一个实施方案中，所述常规无效药物包括或选自抗肿瘤或抗寄生虫的常规无效药物。In one embodiment, the conventional ineffective drugs include or are selected from anti-tumor or antiparasitic conventional ineffective drugs.
在一个实施方案中，所述常规无效药物包括或选自抗肿瘤或抗全身性病原体(寄生虫、细菌、病毒)感染病的常规无效药物。In one embodiment, the conventional ineffective drugs include or are selected from conventional ineffective drugs against tumors or systemic pathogens (parasitic, bacterial, viral) infections.
在本申请公开的范围中，所用术语“常规无效化合物(或药物)”区别于常规有效化合物(例如抗肿瘤化合物)，是指在病原体实验(例如细胞实验)中或许显示出针对特定病原体作用(例如抗肿瘤细胞作用)、但在动物实验中通过吸收作用(常规给药)并不显示出可以实现该抗病原体作用从而有效抑制局部病变疾病(例如实体肿瘤)的化合物。常规无效化合物的常规活性不够高,均未被药政当局(例如FDA)批准用作针对特定局部病变疾病的常规治疗药物(例如抗肿瘤药物)。In the scope of this disclosure, the term "conventional ineffective compound (or drug)" is used to distinguish it from conventional effective compounds (eg, anti-tumor compounds), and means that in pathogen assays (eg, cell assays), it may show an effect against a specific pathogen ( For example, anti-tumor cell effect), but in animal experiments through absorption (conventional administration) does not show that the compound can achieve this anti-pathogen effect to effectively inhibit local disease disease (such as solid tumor). Conventional ineffective compounds are not sufficiently active to be approved by pharmaceutical authorities (such as FDA) as conventional therapeutic drugs (such as antineoplastic drugs) for specific local diseased diseases.
在一个实施方案中，所述局部协同作用所需条件还包括:所述局部协同物选自以下组的一种或多种：氨基酸类营养素、酸消融剂、活体染料、聚多糖,且所述局部协同物以以下给药浓度(重量-体积百分比)(W 局部协同物/V 组合物)进入所述靶区：W 局部协同物/V 组合物可为>0.35％-40％、>0.35％-39％、≥0.5％-40％、≥0.5％-39％、0.5％-40％、0.5％-39％、1％-40％、或1％-39％。，优选可为(3-30)/2.0，更优选可为(4-30)/(2.0-35)或(5-20)/(4.0-30)。 In one embodiment, the conditions required for the topical synergy further include: the topical synergist is one or more selected from the group consisting of amino acid nutrients, acid ablating agents, vital dyes, polysaccharides, and the Topical synergist enters the target area at the following dosing concentrations (weight-volume percent) (W topical synergist /V composition ): W topical synergist /V composition can be >0.35%-40%, >0.35% -39%, ≥0.5%-40%, ≥0.5%-39%, 0.5%-40%, 0.5%-39%, 1%-40%, or 1%-39%. , preferably (3-30)/2.0, more preferably (4-30)/(2.0-35) or (5-20)/(4.0-30).
在本申请公开中，所用术语“氨基酸类营养素”是指药学上可以接受的具有营养保健效应的氨基酸类化合物，优选为选自具有营养保健效应的氨基酸、氨基酸聚合物及氨基酸衍生物，更优选为选自中国、美国或欧洲官方药典或指南所载的氨基酸类营养药和具有营养保健作用的氨基酸类辅料。在本发明的范围中，所用术语“营养保健效应”是指由以下之一种或多种生物作用产生的体内效应：提供能量、参与生物活性物质(例如蛋白质)的合成、参与部分新陈代谢、维持动物肠道微生态平衡、以及参与有利于机体健康的其它生理调节(例如调节蛋白质合成、调节免疫反应)。In the disclosure of the present application, the term "amino acid nutrients" refers to pharmaceutically acceptable amino acid compounds with nutritional health effects, preferably selected from amino acids, amino acid polymers and amino acid derivatives with nutritional health effects, more preferably It is selected from the amino acid nutraceuticals and amino acid excipients with nutritional and health care functions contained in the official pharmacopoeia or guideline of China, the United States or Europe. In the context of the present invention, the term "nutraceutical effect" is used to refer to an in vivo effect resulting from one or more of the following biological actions: providing energy, participating in the synthesis of biologically active substances (eg proteins), participating in partial metabolism, maintaining Animal intestinal microecological balance, and participate in other physiological regulation beneficial to body health (eg regulation of protein synthesis, regulation of immune response).
在本发明的范围内，作为所述氨基酸类营养素的氨基酸、氨基酸聚合物及氨基酸衍生物优选为选自以下组中的氨基酸、或者包含以下组中的氨基酸的氨基酸聚合物、或者以下组中的氨基酸或者包含以下组中的氨基酸的氨基酸聚合物的氨基酸衍生物：蛋白氨基酸和非蛋白氨基酸。Within the scope of the present invention, amino acids, amino acid polymers and amino acid derivatives as the amino acid-based nutrients are preferably amino acids selected from the group consisting of amino acids, or amino acid polymers containing amino acids from the group below, or amino acids selected from the group below. Amino acid derivatives of amino acids or amino acid polymers comprising amino acids from the group of protein amino acids and non-protein amino acids.
在本申请公开的范围中，术语“蛋白氨基酸”是指构成蛋白质的主要氨基酸；而术语“非蛋白氨基酸”是指蛋白氨基酸之外的、也可在营养保健品、传统饮食及功能性饮食(例如保 健饮食)中用作营养保健功能组分的其它氨基酸。In the scope of this disclosure, the term "protein amino acid" refers to the main amino acids that make up protein; and the term "non-protein amino acid" refers to protein amino acids other than protein amino acids, which can also be found in nutraceuticals, traditional diets and functional diets ( For example, other amino acids used as nutritional health functional components in health diets).
具体而言，在本申请公开中，所述蛋白氨基酸包括选自以下组中的氨基酸：非极性氨基酸(例如丙氨酸、缬氨酸、亮氨酸、异亮氨酸、苯丙氨酸、脯氨酸)，极性中性氨基酸(例如色氨酸、酪氨酸、丝氨酸、半胱氨酸、蛋氨酸、天冬酰胺、谷氨酰胺、苏氨酸)，碱性氨基酸(例如赖氨酸、精氨酸、组氨酸)，酸性氨基酸(例如天冬氨酸、谷氨酸)。以上除甘氨酸以外均为L型α-氨基酸。所述非蛋白氨基酸可包括以下氨基酸：β-丙氨酸、牛磺酸、γ-氨基丁酸(GABA)、茶多酚(茶氨酸)、南瓜子氨基酸(3-氨基-3-羧基吡烷酸)、谷氨酰胺、瓜氨酸、鸟氨酸等。Specifically, in the present disclosure, the protein amino acids include amino acids selected from the group consisting of non-polar amino acids (eg, 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 (eg aspartic acid, glutamic acid). All of the above except glycine are L-type α-amino acids. The non-protein amino acids may include the following amino acids: β-alanine, taurine, γ-aminobutyric acid (GABA), tea polyphenols (theanine), pumpkin seed amino acid (3-amino-3-carboxypyridine) alkanoic acid), glutamine, citrulline, ornithine, etc.
在本申请公开的范围中，术语“活体染料”是指药学上可以接受的(因而已用于人体内)能够使靶区有选择性的将特定波长的光吸收或反射的芳香化合物，其可以例如包括亚甲蓝类活体染料。In the context of this disclosure, the term "vital dye" refers to a pharmaceutically acceptable (and thus already used in the human body) aromatic compound capable of selectively absorbing or reflecting light of a specific wavelength at a target area, which can Examples include methylene blue vital dyes.
在本申请公开的范围中，所用术语“酸消融剂”区别于酸碱调节剂，是指在本发明的组合物浓度下局部用药显示出化学消融效应(例如荷瘤动物的抑瘤率在25％以上)的酸性化合物，而这些酸性化合物以较低的浓度用药则并不是作为化学消融剂、而是作为pH等性质的酸碱调节剂。In the scope of this disclosure, the term "acid-ablating agent" is used to distinguish it from acid-base modulators, and means that topical administration at the concentration of the composition of the present invention exhibits a chemical ablative effect (eg, tumor-bearing animals have a tumor inhibition rate of 25%). % or more) acidic compounds, and these acidic compounds are not used as chemical ablation agents at lower concentrations, but as acid-base regulators with properties such as pH.
在本申请公开的范围中，所述酸消融剂为其单药在所用浓度下局部用药可显示出化学消融效应(例如荷瘤动物的抑瘤率在25％以上)、而常规用药则除提供酸碱性外通常并不引入特殊的生物活性的酸性化合物。In the scope of the disclosure of the present application, the acid ablative agent is a single drug that can show chemical ablation effect (for example, the tumor inhibition rate of tumor-bearing animals is more than 25%) at the concentration used, while the conventional drug is not provided In addition to acid and alkali, no special biologically active acidic compounds are usually introduced.
在一个实施方案中，所述局部药物组合物包含免疫球蛋白，其中所述免疫球蛋白浓度(w/v)可为≥2.0、2.0-35％，优选地3.0-30％，更优选地4％至30％，甚至更优选地4-20％，更优选地如3％-10％、4％-10％、3％-8％、4％-8％。In one embodiment, the topical pharmaceutical composition comprises immunoglobulins, wherein the immunoglobulin concentration (w/v) may be ≥ 2.0, 2.0-35%, preferably 3.0-30%, more preferably 4 % to 30%, even more preferably 4-20%, more preferably such as 3%-10%, 4%-10%, 3%-8%, 4%-8%.
任选地，所述局部药物组合物包含一种或多种氨基酸类营养素。Optionally, the topical pharmaceutical composition comprises one or more amino acid nutrients.
任选地，所述局部药物组合物包含一种或多种活体染料。Optionally, the topical pharmaceutical composition comprises one or more vital dyes.
任选地，所述局部药物组合物包含一种或多种酸消融剂。Optionally, the topical pharmaceutical composition comprises one or more acid ablating agents.
在一个实施方案中，所述局部药物组合物包含免疫球蛋白、一种或多种氨基酸类营养素和任选地一种或多种酸消融剂。在一个实施方案中，所述免疫球蛋白在所述药物组合物中的给药浓度(w/v)可为≥2.0、2.0-35％，优选地3.0-30％，更优选地4％至30％，更优选地如4-20％、3％-10％、3％-8％、4％-10％。In one embodiment, the topical pharmaceutical composition comprises immunoglobulin, one or more amino acid based nutrients, and optionally one or more acid ablative agents. In one embodiment, the administration concentration (w/v) of the immunoglobulin in the pharmaceutical composition may be ≥ 2.0, 2.0-35%, preferably 3.0-30%, more preferably 4% to 30%, more preferably such as 4-20%, 3%-10%, 3%-8%, 4%-10%.
在一个实施方案中，所述局部药物组合物包含一种或多种氨基酸类营养素和一种或多种酸消融剂。In one embodiment, the topical pharmaceutical composition comprises one or more amino acid nutrients and one or more acid ablating agents.
在一个实施方案中，所述局部药物组合物包含一种或多种氨基酸类营养素和一种或多种活体染料。In one embodiment, the topical pharmaceutical composition comprises one or more amino acid nutrients and one or more vital dyes.
在一个实施方案中，所述局部药物组合物包含一种或多种酸消融剂和一种或多种活体 染料。In one embodiment, the topical pharmaceutical composition comprises one or more acid ablating agents and one or more vital dyes.
在一个实施方案中，所述局部药物组合物包含一种或多种氨基酸类营养素、一种或多种酸消融剂、和一种或多种活体染料。In one embodiment, the topical pharmaceutical composition comprises one or more amino acid nutrients, one or more acid ablating agents, and one or more vital dyes.
在一个实施方案中，所述氨基酸类营养素在所述药物组合物中的给药浓度(w/v)可为>7％、≥8％、优选可为8-30％，更优选可为10-30％或10％-20％，最优选可为7.5％-15％。In one embodiment, the administration concentration (w/v) of the amino acid nutrients in the pharmaceutical composition may be >7%, ≥8%, preferably 8-30%, more preferably 10% -30% or 10%-20%, most preferably 7.5%-15%.
在一个实施方案中，所述活体染料在所述药物组合物中的给药浓度(w/v)可为>0.3％、≥0.35％、0.5-30％，优选为可0.35-10％或1％-5％，更优选可为0.5-3％或1％-3％。In one embodiment, the administration concentration (w/v) of the vital dye in the pharmaceutical composition may be >0.3%, ≥0.35%, 0.5-30%, preferably 0.35-10% or 1 %-5%, more preferably 0.5-3% or 1%-3%.
在一个实施方案中，所述酸消融剂在所述药物组合物中的给药浓度(w/v)可为≥0.5％，优选可为1.25％-10％，如1.5％-10％，2％-10％，2.5％-10％，2％-7.5％、或2.5％-5％。In one embodiment, the administration concentration (w/v) of the acid ablating agent in the pharmaceutical composition may be ≥ 0.5%, preferably 1.25%-10%, such as 1.5%-10%, 2 %-10%, 2.5%-10%, 2%-7.5%, or 2.5%-5%.
在一个实施方案中，所述氨基酸类营养素选自碱性氨基酸类营养素和/或非碱性氨基酸类营养素之一种或多种。In one embodiment, the amino acid-based nutrients are selected from one or more of basic amino acid-based nutrients and/or non-basic amino acid-based nutrients.
在一个实施方案中，所述局部药物组合物包含所述氨基酸类营养，其中所述氨基酸类营养素包括选自以下碱性氨基酸类营养素之一种或多种:精氨酸、赖氨酸、组氨酸。In one embodiment, the topical pharmaceutical composition comprises the amino acid nutrient, wherein the amino acid nutrient comprises one or more selected from the group consisting of the following basic amino acid nutrient: arginine, lysine, group amino acid.
在一个实施方案中，所述局部药物组合物包含所述氨基酸类营养，其中所述氨基酸类营养素包括选自以下非碱性氨基酸类营养素组之一种或多种：中性氨基酸、酸性氨基酸、氨基酸盐，其中所述中性氨基酸例如：甘氨酸、色氨酸、酪氨酸、丝氨酸、半胱氨酸、蛋氨酸、天冬酰胺、谷氨酰胺、苏氨酸、丙氨酸、缬氨酸、亮氨酸、异亮氨酸、苯丙氨酸、脯氨酸，所述酸性氨基酸例如天冬氨酸、谷氨酸，所述氨基酸盐包括如上所述之氨基酸与酸所形成的盐，例如赖氨酸盐酸盐、组氨酸盐酸盐、谷氨酸盐酸盐、半胱氨酸盐酸盐、精氨酸盐酸盐、硫酸甘氨酸、硫酸甘氨酸铁、赖氨酸盐酸盐、天冬氨酸盐酸盐。In one embodiment, the topical pharmaceutical composition comprises the amino acid nutrient, wherein the amino acid nutrient comprises one or more selected from the group of non-basic amino acid nutrients: neutral amino acids, acidic amino acids, Amino acid salts, wherein the neutral amino acids such as: glycine, tryptophan, tyrosine, serine, cysteine, methionine, asparagine, glutamine, threonine, alanine, valine, Leucine, isoleucine, phenylalanine, proline, the acidic amino acids such as aspartic acid, glutamic acid, and the amino acid salts include the salts formed by the above-mentioned amino acids and acids, such as Lysine hydrochloride, histidine hydrochloride, glutamic acid hydrochloride, cysteine hydrochloride, arginine hydrochloride, glycine sulfate, iron glycinate sulfate, lysine hydrochloride, Aspartate hydrochloride.
在一个实施方案中，所述局部药物组合物包含所述氨基酸类营养，其中所述氨基酸类营养素包括选自以下组中的氨基酸或其盐或者包含或由以下氨基酸构成的寡肽和多肽之一种或多种:精氨酸、赖氨酸、甘氨酸、半胱氨酸、丙氨酸、丝氨酸、谷氨酸。在一个实施方案中，所述寡肽可包括谷胱甘肽。In one embodiment, the topical pharmaceutical composition comprises the amino acid nutrient, wherein the amino acid nutrient comprises an amino acid or a salt thereof selected from the group consisting of or one of oligopeptides and polypeptides comprising or consisting of the following amino acids One or more: arginine, lysine, glycine, cysteine, alanine, serine, glutamic acid. In one embodiment, the oligopeptide may include glutathione.
在一个实施方案中，所述氨基酸类营养素为精氨酸，且所述精氨酸在所述药物组合物中的给药浓度(w/v)≥10％、优选为10-30％，更优选为15％-20％。In one embodiment, the amino acid nutrient is arginine, and the administration concentration (w/v) of the arginine in the pharmaceutical composition is ≥10%, preferably 10-30%, more Preferably it is 15%-20%.
在一个实施方案中，所述局部药物组合物包含所述氨基酸类营养，其中所述氨基酸类营养素包括甘氨酸。In one embodiment, the topical pharmaceutical composition comprises the amino acid nutrient, wherein the amino acid nutrient comprises glycine.
在一个实施方案中，所述药物组合物包含甘氨酸，且所述甘氨酸在所述药物组合物中的给药浓度(w/v)≥10％、优选为10-30％，更优选为10％-15％。In one embodiment, the pharmaceutical composition comprises glycine, and the administration concentration (w/v) of the glycine in the pharmaceutical composition is ≥ 10%, preferably 10-30%, more preferably 10% -15%.
在一个实施方案中，所述药物组合物包含赖氨酸，且所述赖氨酸在所述药物组合物中的给药浓度(w/v)≥10％、优选为10-30％，更优选为10-20％，如15％。In one embodiment, the pharmaceutical composition comprises lysine, and the administration concentration (w/v) of the lysine in the pharmaceutical composition is ≥ 10%, preferably 10-30%, more It is preferably 10-20%, such as 15%.
在一个实施方案中，所述局部药物组合物包含多种所述氨基酸类营养，且所述多种氨基酸类营养素在所述药物组合物中的给药浓度(w/v)可为≥10％、优选可为10-30％，更优选可为10-20％，如10％-15％或15％-20％。In one embodiment, the topical pharmaceutical composition comprises a plurality of the amino acid nutrients, and the administration concentration (w/v) of the plurality of amino acid nutrients in the pharmaceutical composition may be ≥ 10% , preferably can be 10-30%, more preferably can be 10-20%, such as 10%-15% or 15%-20%.
在一个实施方案中，所述局部药物组合物包含所述活体染料，其中所述活体染料优选为为选自以下化合物及其衍生物之一种或多种：亚甲蓝、专利蓝、异硫蓝。In one embodiment, the topical pharmaceutical composition comprises the vital dye, wherein the vital dye is preferably one or more selected from the following compounds and derivatives thereof: methylene blue, patent blue, isosulfide blue.
在一个实施方案中，所述局部药物组合物包含所述活体染料，其中所述活体染料包括亚甲蓝，且其中所述亚甲蓝的给药浓度(w/v)可为0.5％-5.0％、优选可为1％-5％，更优选可为1-3％或1-2％，例如1％、2％或3％。In one embodiment, the topical pharmaceutical composition comprises the vital dye, wherein the vital dye comprises methylene blue, and wherein the methylene blue is administered at a concentration (w/v) of 0.5% to 5.0 %, preferably can be 1%-5%, more preferably can be 1-3% or 1-2%, for example 1%, 2% or 3%.
在一个实施方案中，所述药物组合物包含亚甲蓝和所述氨基酸类营养素。In one embodiment, the pharmaceutical composition comprises methylene blue and the amino acid nutrient.
在一个实施方案中，所述药物组合物包含亚甲蓝和聚多糖。在一个实施方式中，所述聚多糖浓度(w/v)为1-20％,优选为1.5-20％，更优选为1.5-15％，例如10％，或15％。In one embodiment, the pharmaceutical composition comprises methylene blue and polysaccharide. In one embodiment, the polysaccharide concentration (w/v) is 1-20%, preferably 1.5-20%, more preferably 1.5-15%, such as 10%, or 15%.
在一个实施方案中，所述聚多糖包括或选自以下一种或多种：葡聚糖，如羧甲基葡聚糖和水溶性β-D-葡聚糖、右旋糖酐。在一个实施方式中，所述羧甲基葡聚糖的浓度(w/v)可以为10％-15％，如10％；所述水溶性β-D-葡聚糖浓度可以为10％-15％，如15％；所述右旋糖酐的的浓度(w/v)可以为1.5％-10％，如1.5％.In one embodiment, the polysaccharide comprises or is selected from one or more of the following: dextran, such as carboxymethyl dextran and water-soluble beta-D-glucan, dextran. In one embodiment, the concentration (w/v) of the carboxymethyl glucan may be 10%-15%, such as 10%; the concentration of the water-soluble β-D-glucan may be 10%- 15%, such as 15%; the concentration (w/v) of the dextran can be 1.5%-10%, such as 1.5%.
在一个实施方式中，所述药物组合物包含免疫球蛋白、亚甲蓝和任选地聚多糖，其中聚多糖浓度(w/v)可为1-20％,优选可为1.5-20％，更优选为可1.5-15％，例如10％，或15％。In one embodiment, the pharmaceutical composition comprises immunoglobulin, methylene blue and optionally polysaccharide, wherein the polysaccharide concentration (w/v) may be 1-20%, preferably 1.5-20%, More preferably it may be 1.5-15%, such as 10%, or 15%.
在一个实施方案中，所述局部药物组合物包含所述酸消融剂，其中所述酸消融剂选自以下组之一种或多种：弱酸、强酸。In one embodiment, the topical pharmaceutical composition comprises the acid ablating agent, wherein the acid ablating agent is selected from one or more of the following group: weak acid, strong acid.
在一个实施方案中，所述局部药物组合物包含所述酸消融剂，其中所述酸消融剂在所述药物组合物中具有其单药即可显示出化学消融药效的浓度，例如所述弱酸的给药浓度(w/v)可为≥2.0％、≥3.0％或≥5％、优选可为3.0-20％或5-20％，更优选可为5-10％，如7.5％或10％；所述强酸的给药浓度(w/v)可为≥0.5％、≥0.75％或≥1％、优选可为0.5-3％或0.5-3％，如1％或2％。In one embodiment, the topical pharmaceutical composition comprises the acid ablating agent, wherein the acid ablating agent has a concentration in the pharmaceutical composition at which a single agent exhibits chemical ablative efficacy, such as the The weak acid may be administered at a concentration (w/v) of ≥2.0%, ≥3.0% or ≥5%, preferably 3.0-20% or 5-20%, more preferably 5-10%, such as 7.5% or 10%; the administration concentration (w/v) of the strong acid can be ≥0.5%, ≥0.75% or ≥1%, preferably 0.5-3% or 0.5-3%, such as 1% or 2%.
在一个实施方案中，所述弱酸优选为选自以下之一种或多种：碳酸、乙酸、羟基乙酸、丙酸、丙二酸、丁酸、丁二酸、乳酸(2-羟基丙酸)、柠檬酸(2-羟基-1,2,3-丙三羧酸)、苹果酸(2-羟基丁二酸)、酒石酸、草酸、葡萄糖酸；优选为乙酸。In one embodiment, the weak acid is preferably one or more selected from the group consisting of carbonic acid, acetic acid, glycolic acid, propionic acid, malonic acid, butyric acid, succinic acid, lactic acid (2-hydroxypropionic acid) , citric acid (2-hydroxy-1,2,3-propanetricarboxylic acid), malic acid (2-hydroxysuccinic acid), tartaric acid, oxalic acid, gluconic acid; preferably acetic acid.
在一个实施方案中，所述强酸例如包括盐酸、硫酸、硝酸、高氯酸、硒酸、氢溴酸、氢碘酸，优选为盐酸。In one embodiment, the strong acid includes, for example, hydrochloric acid, sulfuric acid, nitric acid, perchloric acid, selenic acid, hydrobromic acid, hydroiodic acid, preferably hydrochloric acid.
在一个实施方案中，所述药物组合物包含所述氨基酸类营养素和弱酸或/和强酸，其中所述氨基酸类营养素包括甘氨酸；所述弱酸包括乙酸；所述强酸包括盐酸。In one embodiment, the pharmaceutical composition comprises the amino acid nutrient and a weak acid or/and a strong acid, wherein the amino acid nutrient comprises glycine; the weak acid comprises acetic acid; and the strong acid comprises hydrochloric acid.
在一个实施方案中，所述药物组合物包含所述氨基酸类营养素、弱酸强碱盐或/和弱酸， 其中所述氨基酸类营养素包括甘氨酸；所述弱酸包括乙酸；所述弱酸强碱盐包括乙酸钠。In one embodiment, the pharmaceutical composition comprises the amino acid nutrient, weak acid and strong base salt or/and weak acid, wherein the amino acid nutrient comprises glycine; the weak acid comprises acetic acid; the weak acid and strong base salt comprises acetic acid sodium.
在一个实施方案中，所述药物组合物包含所述氨基酸类营养素、弱酸强碱盐和活体染料，其中所述氨基酸类营养素包括甘氨酸；所述活体染料包括亚甲蓝；所述弱酸强碱盐包括乙酸钠。In one embodiment, the pharmaceutical composition comprises the amino acid nutrient, a weak acid and strong base salt, and a vital dye, wherein the amino acid nutrient comprises glycine; the vital dye comprises methylene blue; the weak acid strong base salt Includes sodium acetate.
在一个实施方案中，所述药物组合物包含所述氨基酸类营养素、弱酸和活体染料，其中所述氨基酸类营养素包括甘氨酸；所述活体染料包括亚甲蓝；所述弱酸包括乙酸。In one embodiment, the pharmaceutical composition comprises the amino acid nutrient, a weak acid, and a vital dye, wherein the amino acid nutrient comprises glycine; the vital dye comprises methylene blue; and the weak acid comprises acetic acid.
根据本申请公开的药物组合物可以是可包含活性成分(所述药物组合物)的任何适用于局部给药的剂型，优选为以下剂型：注射剂(优选为局部注射剂)、外用液剂、雾化剂等。在一个实施方案中，所述药物组合物为注射用粉针剂。在一个实施方案中，所述药物组合物为液体注射剂。The pharmaceutical composition disclosed according to the present application can be any dosage form suitable for topical administration that can contain the active ingredient (the pharmaceutical composition), preferably the following dosage forms: injection (preferably local injection), external solution, atomization agent, etc. In one embodiment, the pharmaceutical composition is a powder for injection. In one embodiment, the pharmaceutical composition is a liquid injection.
在本发明的范围中，所用术语“注射剂”是指含活性成分和液体载体并供体内给药的无菌制剂。所述注射剂按给药方式分为局部注射剂、静脉注射剂等，静脉注射剂只有在给定局部给药浓度后方可作为局部注射剂使用。注射剂按商品形式分为液体注射剂、注射用粉针剂等。注射用粉针剂包含无菌干粉和溶媒，无菌干粉中包含部分或全部活性成分，溶媒中包含全部液体载体。注射剂中所述活性成分的浓度均为其与全部所述液体载体的混合物中的活性成分浓度，通常是局部给药器械(注射器、穿刺器、灌注导管等)终点(例如针孔、导管出口等)的液体药物中的活性成分浓度。对注射用粉针剂而言，所述活性成分的浓度即为无菌干粉和溶媒的混合物(例如复溶液，或所述药物学可接受的液体载体)中的活性成分浓度。In the context of the present invention, the term "injectable" is used to refer to a sterile preparation containing the active ingredient and a liquid carrier for in vivo administration. The injections are classified into local injections, intravenous injections, etc. according to the mode of administration, and the intravenous injections can be used as local injections only after a given local administration concentration. Injections are classified into liquid injections, powder injections, etc. A powder for injection contains sterile dry powder and a vehicle, the sterile dry powder contains part or all of the active ingredients, and the vehicle 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, usually the end point (such as needle hole, catheter outlet, etc.) of the local drug delivery device (syringe, trocar, perfusion catheter, etc.) ) of the active ingredient concentration in the liquid medicine. For injection powders, the concentration of the active ingredient is the concentration of the active ingredient in a mixture of sterile dry powder and a vehicle (eg, a reconstituted solution, or the pharmaceutically acceptable liquid carrier).
在本发明的范围中，术语“外用液剂”是指含活性成分和液体载体并供体表[例如皮肤、粘膜(例如眼粘膜、鼻粘膜等)或/和腔道(例如口腔、直肠、***、尿道、鼻腔、耳道等)]给药的液体药物，其包括例如洗剂、搽剂、滴剂、含漱剂、涂剂等。局部给药时，所述液体药物往往来自洗液瓶、滴液瓶、滴液管、潄洗液瓶、棉签等局部给药器械。所述外用液剂中所述活性成分的浓度即该液体药物中的活性成分浓度。In the context of the present invention, the term "liquid for external use" refers to an active ingredient and a liquid carrier and a donor surface [e.g. skin, mucous membranes (e.g. ocular mucosa, nasal mucosa, etc.) or/and cavities (e.g. oral cavity, rectum, Vaginal, urethral, nasal, ear canal, etc.)] administration of liquid medicines, which include, for example, lotions, liniments, drops, gargles, paints, and the like. In the case of topical administration, the liquid medicine often comes from local administration devices such as lotion bottles, drip bottles, drip tubes, rinse bottles, cotton swabs, and the like. The concentration of the active ingredient in the external liquid preparation is the concentration of the active ingredient in the liquid medicine.
在本发明的范围中，术语“雾化剂”是指含活性成分和液体载体并在使用时可借助压力将上述液体药物雾化喷出给药的剂型，其可供皮肤、粘膜(例如眼粘膜、鼻粘膜等)或/和腔道(例如口腔、直肠、***、尿道、鼻腔、耳道等)给药，其包括例如气雾剂、喷雾剂、雾化剂等。局部给药时，液体药物的雾化往往通过气雾器、雾化器、喷雾器等局部给药器械来形成。药物雾化喷出至目标位后，又累积形成液体药物。该液体药物与雾化前的液体药物在组成上大致相同。因而，雾化剂中所述活性成分的浓度可以用雾化前液体药物中的活性成分浓度来代表。In the context of the present invention, the term "nebulizer" refers to a dosage form which contains the active ingredient and a liquid carrier and which, in use, can be administered by atomizing the aforementioned liquid drug by means of pressure, which can be applied to the skin, mucous membranes (e.g. ocular mucosa, nasal mucosa, etc.) or/and cavity (eg, oral, rectal, vaginal, urethral, nasal, ear canal, etc.) administration, including, eg, aerosols, sprays, nebulizers, and the like. During topical administration, the atomization of liquid drugs is often formed by topical drug delivery devices such as aerosols, atomizers, and nebulizers. After the drug is atomized and sprayed to the target position, it accumulates to form a liquid drug. The liquid medicine is substantially the same in composition as the liquid medicine before atomization. Thus, the concentration of the active ingredient in the nebulizer can be represented by the concentration of the active ingredient in the liquid drug prior to nebulization.
本领域技术人员会理解，根据本发明的技术方案，本发明的药物组合物应被制成可向所述靶区局部给药的剂型。Those skilled in the art will understand that, according to the technical solutions of the present invention, the pharmaceutical composition of the present invention should be made into a dosage form that can be locally administered to the target area.
根据本发明的制备方法，本发明的药物组合物的制备包含以下步骤：制备含所述必须 组分(例如所述药物组合物)和任选存在的其它物质的液体药物。该液体药物可以是溶液(例如为亲水溶媒的溶液、优选为水溶液)、悬浮液、或乳浊液。当所述液体药物为悬浊液时，其中的分散介质可以本领域技术人员已知的任意合适者，例如微米材料或纳米材料。当所述液体药物为乳浊液时，其中的分散介质可以本领域技术人员已知的任意合适者，例如可用于注射的植物油、合成油或半合成油。其中所述植物油可以是例如棉籽油、杏仁油、橄榄油、蓖麻油、芝麻油、大豆油和花生油。According to the preparation method of the present invention, the preparation of the pharmaceutical composition of the present invention comprises the steps of preparing a liquid medicine containing the essential components (such as the pharmaceutical composition) and optionally other substances. The liquid drug can be a solution (eg, in a hydrophilic vehicle, preferably an aqueous solution), a suspension, or an emulsion. When the liquid drug is a suspension, the dispersion medium therein can be any suitable one known to those skilled in the art, such as micro-materials or nano-materials. When the liquid drug is an emulsion, 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 for injection. Wherein the vegetable oil may be, for example, cottonseed oil, almond oil, olive oil, castor oil, sesame oil, soybean oil and peanut oil.
根据本申请公开的药物组合物可以如此制备或组成,以致于其中包含的所述Ig或/和其共用物的浓度可以大于或等于其用以满足所述局部协同作用所需的给药浓度。当大于该给药浓度时，可进一步被稀释使用。The pharmaceutical compositions disclosed in accordance with the present application can be prepared or composed such that the Ig or/and its co-conjugates can be contained in a concentration greater than or equal to its administration concentration required to satisfy the local synergistic effect. When it is greater than the administration concentration, it can be further diluted for use.
根据本发明制备方法的一个实施方案，本发明的药物组合物液体注射剂可通过包含以下步骤的方法制备：1)将根据局部给药浓度所需量的必须组分(例如所述药物组合物)以及任选存在的其他组分加入水中制备为液体；2)将根据局部给药浓度所需量的任选存在的其它药物加入在1)中制备的液体混合均匀获得液体药物；3)将在2)中制备的液体药物除菌后制成液体注射剂。使用时，液体注射剂中的除菌液体药物可直接或稀释后用作局部给药液体药物。According to one embodiment of the preparation method of the present invention, the liquid injection of the pharmaceutical composition of the present invention can be prepared by a method comprising the following steps: 1) The necessary components (eg, the pharmaceutical composition) in the required amount according to the local administration concentration And other components optionally present are added into water to prepare a liquid; 2) the other drugs optionally present according to the required amount of local administration concentration are added to the liquid prepared in 1) and mixed uniformly to obtain a liquid drug; 3) will be in the The liquid medicine prepared in 2) was sterilized to make a liquid injection. In use, the sterilized liquid medicine in the liquid injection can be used as a liquid medicine for topical administration directly or after dilution.
根据本申请公开的一个实施方案，本发明的药物组合物之注射用粉针剂可通过包括以下步骤的方法制备：制备含根据局部给药浓度所需量的所述药物组合物的无菌干粉；和制备包含根据局部给药浓度所需量的所述任选存在的其他组分的无菌溶媒。所述除菌干粉优选为除菌冻干干粉，其制备方法包括：1)制备包含氨基酸来营养素、水溶性弱中和物质以及任选存在的其他组分的溶液；2)除菌过滤和分装；3)冷冻干燥；4)压塞、轧盖。所述冷冻干燥的工艺条件例如包括：预冻条件为在预冻温度-45℃保持4小时；升华干燥条件为升温速率为0.1℃/分钟、且升至-15℃时至少保持10小时；解吸附干燥条件为30℃保持6小时。使用时，将注射用粉针剂的无菌干粉复溶于无菌溶媒中形成复溶液体药物，其即可直接或稀释后用作局部给药液体药物。According to one embodiment disclosed in the present application, the injection powder of the pharmaceutical composition of the present invention can be prepared by a method comprising the following steps: preparing a sterile dry powder containing the pharmaceutical composition in the required amount according to the local administration concentration; and preparation of sterile vehicles containing the required amounts of the optional other components according to the concentration for topical administration. Described sterilization dry powder is preferably sterilization freeze-dried dry powder, and its preparation method comprises: 1) prepare the solution that comprises amino acid to nutrient, water-soluble weak neutralization substance and optional other components; 2) sterilization filtration and separation; 3) freeze-drying; 4) plugging and capping. The freeze-drying process conditions include, for example: the pre-freezing conditions are kept at the pre-freezing temperature -45°C for 4 hours; the sublimation drying conditions are that the heating 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 were kept at 30°C for 6 hours. When in use, the sterile dry powder of the powder for injection is reconstituted in a sterile solvent to form a reconstituted liquid drug, which can be used directly or after dilution as a liquid drug for topical administration.
根据本发明制备方法的一个实施方案，本发明的药物组合物外用液剂通过包含以下步骤的方法制备：将根据局部给药浓度所需量的药物组合物以及任选存在的其他组分加入溶媒中制备为液体药物。使用时，外用液剂中的液体药物可直接或稀释后用作局部给药液体药物。According to one embodiment of the preparation method of the present invention, the external use liquid of the pharmaceutical composition of the present invention is prepared by a method comprising the following steps: adding the required amount of the pharmaceutical composition according to the local administration concentration and optionally other components into a vehicle It is prepared as a liquid medicine. In use, the liquid medicine in the external solution can be used directly or after being diluted as a liquid medicine for topical administration.
根据本发明制备方法的一个实施方案，本发明的药物组合物雾化剂可通过包含以下步骤的方法制备：1)将根据局部给药浓度所需量的药物组合物以及雾化赋形剂加入溶媒中制备为液体；2)将根据局部给药浓度所需量的任选存在的其他组分加入在1)中制备的液体中混合均匀获得液体药物。常用的雾化赋形剂包括例如：甘油、聚山梨酯-80、苯扎氯铵、微晶纤维素-羧甲基纤维素钠等。使用时，该液体药物加入雾化器(例如喷雾剂)并在其雾化作用下以雾滳形式局部给药至靶区，这些雾滳在靶区累积为液体药物。According to one embodiment of the preparation method of the present invention, the pharmaceutical composition aerosol of the present invention can be prepared by a method comprising the following steps: 1) adding the required amount of the pharmaceutical composition according to the local administration concentration and aerosol excipients into The vehicle is prepared as a liquid; 2) the other components optionally present in the required amount according to the local administration concentration are added to the liquid prepared in 1) and mixed uniformly to obtain a liquid medicine. Commonly used excipients for nebulization include, for example: glycerin, polysorbate-80, benzalkonium chloride, microcrystalline cellulose-sodium carboxymethyl cellulose, and the like. In use, the liquid drug is added to a nebulizer (eg, a spray) and is locally administered to the target area in the form of mist under its atomization, and the mist is accumulated in the target area as a liquid drug.
根据本申请公开的再一个方面，其提供一种用于治疗局部病变疾病的装置，其包含根据本申请公开所述的药物组合物。在一个实施方案中，所述装置包括气雾器、雾化器、喷雾器。According to yet another aspect of the present disclosure, there is provided a device for treating local pathological diseases, comprising the pharmaceutical composition according to the present disclosure. In one embodiment, the device comprises an aerosol, nebulizer, nebulizer.
按上述这些方法的原则，本领域技术人员可以采用任意合适的具体方法制备多种包含本发明组合物的具体剂型。例如，本发明的药物组合物中的变化包括：含不同种类和浓度的所述药物组合物、含不同种类和浓度的其它药物、含不同种类和浓度的其他添加剂(例如止痛剂、增活剂等)。Following the principles of these methods described above, one skilled in the art can employ any suitable specific method to prepare a variety of specific dosage forms comprising the compositions of the present invention. For example, variations in the pharmaceutical compositions of the present invention include: containing different types and concentrations of the pharmaceutical composition, containing different types and concentrations of other drugs, containing different types and concentrations of other additives (eg, analgesics, activators, etc.) Wait).
在本申请公开中，所述药物组合物主要是用于通过局部给药预防和治疗局部病变疾病、尤其是难治性局部病变疾病。In the disclosure of the present application, the pharmaceutical composition is mainly used for the prevention and treatment of local pathological diseases, especially refractory local pathological diseases, by topical administration.
在本发明的范围中，术语“局部病变疾病”是指具有局部病变症状的疾病，而术语“局部病变”是指动物(优选人类)身体局部部位原生或继生的结构、形态或功能上的不正常，其例如可以是包括以下一种或多种：瘤体、非瘤肿大、局部炎症、分泌腺分泌功能异常等。所述局部部位可以是本领域技术人员已知的任意合适者，例如可以是包括以下一种或多种器官中的局部：分泌***所在的分泌器官、血液循环***所在的心血管器官、皮肤等。In the context of the present invention, the term "focal disease" refers to a disease with symptoms of a local disease, whereas the term "focal disease" refers to a structural, morphological or functional disorder that is native or secondary to a local part of the body of an animal (preferably a human). Abnormal, for example, may include one or more of the following: tumor, non-tumor swelling, local inflammation, abnormal secretion of secretory glands, and the like. The local site can be any suitable one known to those skilled in the art, for example, it can be a local part including one or more of the following organs: the secretory organ where the secretory system is located, the cardiovascular organ where the blood circulatory system is located, the skin, etc. .
局部给药要求药物组成(局部活性成分、组成比及组分浓度)可通过介入手段给药于给局部病变所在的组织、并在该组织内产生所需要的疗效。例如，当病变为肿瘤时，局部组织为肿瘤细胞所在的瘤体；当病变为非瘤肿大时，局部组织为诸如肿块的异常，例如增生、囊肿、结节等病变肿块；当病变为局部炎症时，局部组织为发炎区，例如发炎肿大体；当病变为分泌异常时，局部组织为异常源或其所在的分泌腺体。又例如，当疾病为胰岛素分泌异常时，异常源在胰岛，局部组织则为胰岛或胰岛所在的胰腺；当疾病为皮肤病时，局部组织为病变皮肤或病变皮肤的附属器官。Local administration requires that the drug composition (local active ingredient, composition ratio and component concentration) can be administered to the tissue where the local lesion is located by interventional means, and produce the desired therapeutic effect in the tissue. For example, when the lesion is a tumor, the local tissue is the tumor body where the tumor cells are located; when the lesion is non-tumor, the local tissue is an abnormality such as a mass, such as hyperplasia, cyst, nodule, etc.; when the lesion is a local In the case of inflammation, the local tissue is the inflamed area, such as an inflamed body; when the lesion is abnormal secretion, the local tissue is the abnormal source or the secretory gland where it is located. For another example, when the disease is abnormal insulin secretion, the abnormal source is the pancreatic islet, and the local tissue is the pancreatic islet or the pancreas where the pancreatic islet is located; when the disease is a skin disease, the local tissue is the diseased skin or an accessory organ of the diseased skin.
具体而言，在本申请公开中，所述靶区组织为包含癌细胞或/和成纤维细胞的局部病变组织；所述局部病变包括肿瘤、非瘤肿大、局部炎症、分泌腺功能异常和皮肤病。Specifically, in the disclosure of the present application, the target tissue is a local lesion tissue containing cancer cells or/and fibroblasts; the local lesion includes tumor, non-tumor swelling, local inflammation, abnormal secretion gland function and skin disease.
在本申请公开的范围内，术语“肿瘤”是指由于细胞或变异的细胞异常增殖形成的肿块，其包括实体肿瘤。术语“实体肿瘤”是指具有瘤体的肿瘤，其可以是由于任何病理(恶性和非恶性)和处于任何阶段的肿瘤，包括例如按照肿瘤细胞类型进行分类的以下组：上皮细胞肿瘤、肉瘤、淋巴瘤、生殖细胞肿瘤、胚细胞瘤；以及包括按照肿瘤细胞集中区所在的器官或组织来命名的肿瘤，包括例如按照以下器官或组织来命名的肿瘤：皮肤、骨、肌肉、乳腺、肾、肝、肺、胆囊、胰腺、脑、食道、膀肌、大肠、小肠、脾、胃、***、翠丸、卵巢或子宫。Within the scope of this disclosure, the term "tumor" refers to a mass formed due to abnormal proliferation of cells or mutated cells, including solid tumors. The term "solid tumor" refers to a tumor with a tumor body, which can be due to any pathology (malignant and non-malignant) and at any stage, including, for example, the following groups classified by tumor cell type: epithelial cell tumors, sarcomas, Lymphoma, germ cell tumor, blastoma; and tumors named after the organ or tissue in which the tumor cell concentration is located, including, for example, tumors named after the following organs or tissues: skin, bone, muscle, breast, kidney, Liver, Lung, Gallbladder, Pancreas, Brain, Esophagus, Bladder, Large Intestine, Small Intestine, Spleen, Stomach, Prostate, Green Pill, Ovary or Uterus.
具体而言，所述恶性肿瘤包括例如乳腺癌、胰腺癌、甲状腺癌、鼻咽癌、***癌、肝癌、肺癌、肠癌、口腔癌、食道癌、胃癌、喉癌、睾丸癌、***癌、子宫癌、卵巢癌等。Specifically, the malignant tumor includes, for example, breast cancer, pancreatic cancer, thyroid cancer, nasopharyngeal cancer, prostate cancer, liver cancer, lung cancer, colon cancer, oral cancer, esophageal cancer, gastric cancer, throat cancer, testicular cancer, vaginal cancer, Uterine cancer, ovarian cancer, etc.
所述非恶性肿瘤包括例如乳腺瘤、胰腺瘤、甲状腺瘤、***瘤、肝瘤、肺瘤、肠瘤、口腔瘤、食道瘤、胃瘤、鼻咽瘤、喉瘤、睾丸瘤、***瘤、子宫瘤、输卵管瘤、卵巢瘤等。The non-malignant tumors include, for example, breast tumors, pancreatic tumors, thyroid tumors, prostate tumors, liver tumors, lung tumors, intestinal tumors, oral tumors, esophageal tumors, gastric tumors, nasopharyngeal tumors, laryngeal tumors, testicular tumors, vaginal tumors, Uterine tumor, fallopian tube tumor, ovarian tumor, etc.
在一个实施方案中，所述局部病变疾病包括非瘤肿大。术语“非瘤肿大”是指肿瘤以外的肿大，包括例如增生(例如乳腺、胰腺、甲状腺、甲状旁腺、***等的增生)、囊肿(例如乳腺、甲状腺、甲状旁腺等的囊肿)、结节(例如乳腺、甲状腺、甲状旁腺等的结节)、异常静脉团(例如痔疮等)、局部炎症发肿、微生物感染发肿等。所述痔疮包括内痔、外痔、混合痔。In one embodiment, the locally diseased disease comprises a non-tumorous enlargement. The term "non-neoplastic" refers to enlargements other than tumors, including, for example, hyperplasia (eg, hyperplasia of breast, pancreas, thyroid, parathyroid, prostate, etc.), cysts (eg, cysts of breast, thyroid, parathyroid, etc.) , Nodules (such as nodules of breast, thyroid, parathyroid, etc.), abnormal venous mass (such as hemorrhoids, etc.), local inflammation and swelling, microbial infection and swelling. The hemorrhoids include internal hemorrhoids, external hemorrhoids and mixed hemorrhoids.
在一个实施方案中，所述局部病变疾病包括局部炎症、尤其是难治性炎症。在本发明的范围内，术语“局部炎症”是指局部部位的非瘤性发炎，包括例如变质性炎(alterative inflammation)、渗出性炎(exudative inflammation)和增生性炎，其可以是本领域技术人员已知的任意合适者，例如可以是包括以下一种或多种：关节炎、乳腺炎、胰腺炎、甲状腺炎、***炎、肝炎、肺炎、肠炎、口腔炎、咽炎、牙周炎、食道炎、胃炎、胃溃疡、鼻炎、鼻窦炎、喉炎、气管炎、支气管炎、***炎、子宫炎、输卵管炎、***等。In one embodiment, the locally diseased disease comprises local inflammation, especially refractory inflammation. In the context of the present invention, the term "localized inflammation" refers to non-neoplastic inflammation of a localized site, including, for example, alterative, exudative, and proliferative inflammation, which may be in the art Any suitable known to the skilled person, for example, may include one or more of the following: arthritis, mastitis, pancreatitis, thyroiditis, prostatitis, hepatitis, pneumonia, enteritis, stomatitis, pharyngitis, periodontitis, Esophagitis, gastritis, gastric ulcer, rhinitis, sinusitis, laryngitis, bronchitis, bronchitis, vaginitis, metritis, salpingitis, oophoritis, etc.
在一个实施方案中，所述局部病变疾病包括皮肤病、尤其是难治性皮肤病。在本发明的范围内，术语“皮肤病”是指原生或继生在皮肤或皮肤附属器官的病变，其可以是本领域技术人员已知的任意合适者，例如可以是包括以下一种或多种：皮肤癌、皮肤非恶性肿瘤、病毒性皮肤病(例如疱疹、疣、风疹、手足口病)、细菌性皮肤病(例如脓疱病、疖、麻风)、真菌性皮肤病(例如各种癣)、性传播疾病(例如梅毒、淋病及***)、过敏性与自身免疫性皮肤病(例如接触性皮炎、湿疹、荨麻疹)、物理性皮肤病(例如日光性皮肤病、冻疮、鸡眼、手足皲裂、压疮)、***疾病(例如红斑狼疮)、色素障碍性皮肤病(例如雀斑、色素痣、各种斑)、皮肤附属器疾病(例如痤疮、酒渣鼻、脂溢性皮炎、斑秃、秃发、多汗症及臭汗症)。In one embodiment, the localized disease comprises a skin disease, especially a refractory skin disease. In the context of the present invention, the term "skin disease" refers to a disease that is native or secondary to the skin or skin appendages, which may be any suitable known to those skilled in the art, for example may include one or more of the following Species: skin cancer, non-malignant skin tumors, viral skin diseases (e.g. herpes, warts, rubella, hand, foot and mouth disease), bacterial skin diseases (e.g. impetigo, boils, leprosy), fungal skin diseases (e.g. various ringworm), sexually transmitted diseases (such as syphilis, gonorrhea, and condyloma acuminatum), allergic and autoimmune skin diseases (such as contact dermatitis, eczema, urticaria), physical skin diseases (such as solar dermatosis, frostbite, corns) , chapped hands and feet, pressure ulcers), connective tissue diseases (such as lupus erythematosus), dyspigmented skin diseases (such as freckles, pigmented nevi, various macules), skin adnexal diseases (such as acne, rosacea, seborrheic dermatitis) , alopecia areata, alopecia, hyperhidrosis and stinkyhidrosis).
在一个实施方案中，所述局部病变疾病包括分泌腺分泌功能异。在本发明的范围内，术语“分泌腺”是指由腺细胞或腺细胞群组成的、执行分泌功能(secretion)的结构，其包括外分泌腺和内分泌腺。分泌腺分泌功能异常包括分泌腺功能亢进症(例如甲状腺功能亢进症)和分泌腺功能减退症(例如甲状腺功能減退症、胰岛功能減退症(糖尿病之一种))。In one embodiment, the focal disease comprises secretory gland dysfunction. In the context of the present invention, the term "secretory gland" refers to a structure composed of glandular cells or groups of glandular cells that performs a secretory function, including exocrine and endocrine glands. Abnormal secretion function of secretory glands includes hypersecretory gland function (eg, hyperthyroidism) and hyposecretory gland function (eg, hypothyroidism, islet dysfunction (a type of diabetes)).
在一个实施方案中，所述局部病变疾病包括心血管疾病。介入治疗己经成为心血管疾病的一种重要治疗手段。所述心血管疾病包括例如血管瘤、肥厚型梗阻性心肌病、心房颤动、心律失常、动脉栓塞等。In one embodiment, the localized disease comprises cardiovascular disease. Interventional therapy has become an important treatment for cardiovascular disease. Such cardiovascular diseases include, for example, hemangiomas, hypertrophic obstructive cardiomyopathy, atrial fibrillation, arrhythmias, arterial embolism, and the like.
本申请公开的所述局部药物是一种治疗药物，当其用于预防和治疗局部病变疾病时，还可与其它介入疗法、全身化疗、免疫疗法、光动力疗法、声动力疗法、手术干预或此类疗法的组合相组合施用，以进一步提高疗效。The topical drug disclosed in the present application is a therapeutic drug, and when it is used for the prevention and treatment of local lesions, it can also be combined with other interventional therapy, systemic chemotherapy, immunotherapy, photodynamic therapy, sonodynamic therapy, surgical intervention or Combinations of such therapies are administered in combination to further enhance efficacy.
在本申请公开中，所述药物组合物主要是用于通过局部给药预防和治疗局部病变疾病。In the disclosure of the present application, the pharmaceutical composition is mainly used for the prevention and treatment of local pathological diseases through topical administration.
在根据本申请公开的局部治疗和预防局部病变疾病的应用和方法中，所述一种包含免疫球蛋白及其局部协同物以它们在所述局部药物组合物中的浓度或量比局部给药。该浓度 或量比局部给药能够提供局部反应的协同作用。In the application and method of topical treatment and prevention of local pathological diseases disclosed according to the present application, the one comprising immunoglobulin and its topical synergist is administered locally in the concentration or amount ratio thereof in the topical pharmaceutical composition . This concentration or amount ratio can provide a synergistic effect of the local response when administered locally.
在一个实施方案中，所述方法包括在局部用药所述药物组合物之前、期间或之后还任选进行一种或多种其它治疗，例如化疗、免疫疗法、放射疗法、手术、物理消融。In one embodiment, the method comprises optionally performing one or more other treatments, eg, chemotherapy, immunotherapy, radiation therapy, surgery, physical ablation, before, during, or after topical administration of the pharmaceutical composition.
基于在下文中更详细描述的研究，尽管具体机理尚待进一步研究，本发明的药物组合物显示出促进局部病变所在组织的相关结构(例如病变组织、病变细胞及参与构成它们的任一结构)的有效破坏、同时对患者正常组织仅有最小化的损害，从而达到安全、有效治疗局部病变疾病的药学效果。Based on the studies described in more detail below, although the specific mechanism remains to be further studied, the pharmaceutical composition of the present invention is shown to promote the promotion of local lesions in the relevant structures of the tissue (eg, diseased tissue, diseased cells, and any structure involved in constituting them). It can effectively destroy and at the same time only minimize the damage to the normal tissue of patients, so as to achieve the pharmaceutical effect of safe and effective treatment of local lesions.
实施例Example
通过以下具体实施例对本发明作进一步的说明，但不作为对本发明的限制。在以下实施例中，所有的试验动物均依照相关法规及行业自律进行。如无特殊说明，所有试验均按常规方法进行。The present invention will be further illustrated by the following specific examples, but not intended to limit the present invention. In the following examples, all experimental animals were carried out in accordance with relevant regulations and industry self-discipline. Unless otherwise specified, all tests were carried out by conventional methods.
以下具体实施例所用的材料、试剂等，如无特殊说明，均可从商业途径得到。以下实施例中所用的部分材料列于表1。The materials, reagents, etc. used in the following specific examples can be obtained from commercial sources unless otherwise specified. Some of the materials used in the following examples are listed in Table 1.
表1Table 1
Figure PCTCN2022076823-appb-000001
Figure PCTCN2022076823-appb-000001
以下实施例中所用的实验动物均为通过专业实验动物公司购入，均为SPF(Specific Pathogen Free，无特定病原体)级动物。以小鼠为例，共有2种：BALB/c小鼠、裸小鼠，其中裸小鼠为裸基因(nu)导入BALB/c小鼠获得的突变系(BALB/c–nu)小鼠。小鼠均为6-8周龄健康雌性、体重17.5-20.5g。The experimental animals used in the following examples were purchased from professional experimental animal companies, and all were SPF (Specific Pathogen Free, specific pathogen-free) grade animals. Taking mice as an example, there are two types: BALB/c mice and nude mice. Nude mice are mutant (BALB/c–nu) mice obtained by introducing nude gene (nu) into BALB/c mice. The mice were all healthy females aged 6-8 weeks, weighing 17.5-20.5 g.
在以下实施例中，肿瘤体积(V)、相对肿瘤增殖率(R)、抑瘤率(r)的计算公式分别如下：In the following examples, the calculation formulas of tumor volume (V), relative tumor proliferation rate (R), and tumor inhibition rate (r) are respectively as follows:
肿瘤体积V＝l/2×a×b 2，其中a表示肿瘤长，b表示肿瘤宽。 Tumor volume V=l/2×a×b 2 , where a represents tumor length and b represents tumor width.
体积抑瘤率r(％)＝(CV-TV)/CV×100％，其中TV为研究组的平均体积；CV为阴性对照组的平均体积。Volume tumor inhibition rate r(%)=(CV-TV)/CV×100%, where TV is the average volume of the study group; CV is the average volume of the negative control group.
抑瘤率R(％)＝(CW-TW)/CW×100％，其中TW为研究组的平均瘤重；CW为阴性对照组的平均瘤重。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.
在以下实施例中，药物i的药效记为E i,其中E i可以为(100-r i)％或R i％。在本发明的范围內,多个药物(i＝1、2、…n)的组合物记为药物1/药物2/…/药物n,例如药物A和药物B的组合物记为B/A。A、B的单用药效分别记为E A和E B,A/B的实际共用药效记为E A+B。 In the following examples, the efficacy of drug i is denoted as E i , where E i can be (100-r i )% or R i %. Within the scope of the present invention, the composition of a plurality of drugs (i=1, 2, ... n) is denoted as drug 1/drug 2/.../drug n, eg the composition of drug A and drug B is denoted as B/A . The single drug efficacy of A and B is recorded as EA and EB respectively, and the actual combined efficacy of A/B is recorded as EA +B .
在以下实施例中，试验结果(例如瘤重、瘤体积)采用均数±标准差(x±s)表示，两个试验动物组与组均数之间的差别采用统计学软件SPSS 13.0或SPSS 19.0软件进行显著性检验来比较，检验选用统计量t来进行，检验水准α＝0.05，P<0.05表示差异有统计学意义，P>0.05则无统计学意义。In the following examples, the test results (such as tumor weight, tumor volume) are expressed as mean ± standard deviation (x ± s), and the difference between the two experimental animal groups and the group mean is expressed by statistical software SPSS 13.0 or SPSS 19.0 software was used to carry out a significant test for comparison, and the test was carried out with the statistic t. The test level was α=0.05, P<0.05 indicated that the difference was statistically significant, and P>0.05 was not statistically significant.
在以下实施例中，药物作用类型(共用药理)通过药物效应、尤其是比较同一研究药物在不同技术方案中的药物效应来进行研究。例如,当药物i在方案X和Y之间的药物效应差异并非不寻常地大(例如E iX/E iY<150％)时,其很可能是基本相同的药物作用类型(药理)中不同动力学条件(作用浓度)所致；而当该药物效应差异超乎寻常地大(例如E iX/E iY>150％)时,药物i在方案X中的药物效应之大应当超过了其在方案Y中的药物作用类型(药理)的动力学预期范围,从而很可能涉及与方案Y不同的药物作用类型(药理)。当两个药物显示出显然不同的E iX/E iY关系,则它们很可能涉及显然不同的药理；而当两个药物显示出相似的E iX/E iY关系,则它们很可能涉及相同的药理、至少涉及相似的药理(例如类化学消融药理和化学消融药理)。 In the following examples, the type of drug action (common pharmacology) is studied by drug effect, especially comparing the drug effect of the same investigational drug in different technical protocols. For example, when the difference in drug effect of drug i between regimens X and Y is not uncommonly large (eg, E iX /E iY < 150%), it is likely to be different kinetics in essentially the same type of drug action (pharmacology) and when the drug effect difference is unusually large (eg E iX /E iY >150%), the drug effect of drug i in regimen X should be greater than its The expected range of kinetics for the type of drug action (pharmacology) in Y, and thus likely to involve a different type of drug action (pharmacology) than in regimen Y. When two drugs show clearly different E iX /E iY relationships, they are likely to involve clearly different pharmacology; and when two drugs show similar E iX /E iY relationships, they are likely to involve the same pharmacology , involving at least similar pharmacology (eg chemoablation-like pharmacology and chemoablation pharmacology).
在以下实施例中，全身活性阳性对照物包括经典细胞毒药物(例如0.5-1％5-氟尿嘧啶,其在以下实施例条件下的抑瘤率为≥30％),局部活性阳性对照物包括经典化学消融剂(例如75-99％乙醇,其在以下实施例局部给药条件下的抑瘤率为≥30％)。In the following examples, the systemic active positive controls include classical cytotoxic drugs (such as 0.5-1% 5-fluorouracil, whose tumor inhibition rate is ≥30% under the conditions of the following examples), and the locally active positive controls include classical Chemical ablative agents (eg, 75-99% ethanol, whose tumor inhibition rate is ≥30% under the local administration conditions of the following examples).
药物联合使用产生的药理作用(或药效)具有高度的不确定性，业内往往依据以下实际药效/预期药效比q判断进行：q＝q 1/q 2，其中q 1为实际共用药效，q 2为根据各药物i的实际单用药效(E A、E B、等)得出的预期相加药效。 The pharmacological effects (or efficacy) produced by the combined use of drugs are highly uncertain, and the industry often judges based on the following actual efficacy/expected efficacy ratio q: q=q 1 /q 2 , where q 1 is the actual shared drug effect, q 2 is the expected additive effect based on the actual single effect (EA, EB , etc.) of each drug i .
q 2的计算方法很多,大多针对细胞试验效应。通常认为,当q＝1时,实际共用作用符合理论预期,显示为相加作用；当实际/预期比q<1时,实际共用作用弱于理论预期,显示为拮抗作用；当实际/预期比q>1时,实际共用作用超理论预期,显示为协同作用,且q越大显示其超理论预期越远、或协同作用越强。 There are many calculation methods for q 2 , most of which are aimed at cell test effects. It is generally believed that when q=1, the actual sharing effect is in line with the theoretical expectation, showing an additive effect; when the actual/expected ratio q<1, the actual sharing effect is weaker than the theoretical expectation, showing an antagonistic effect; when the actual/expected ratio When q>1, the actual synergistic effect exceeds the theoretical expectation, showing a synergistic effect, and the larger the q, the farther it exceeds the theoretical expectation, or the stronger the synergy effect.
一种判断动物试验中药物共用效应的方法是Burgi法(Burgi Y.Pharmacology；Drug actions and reactions.Cancer res.1978,38(2),284-285)。金正均对Burgi法进行了改进(金正均，等概率和曲线和“Q50”，上海第二医学院学报；1981,1,75-86),药物作用的单纯相加被认为是互相独立的作用的相加,其q计算式为：A method for judging drug co-effects in animal experiments is the Burgi method (Burgi Y. Pharmacology; Drug actions and reactions. Cancer res. 1978, 38(2), 284-285). Jin Zhengjun improved Burgi's method (Jin Zhengjun, equal probability and curve and "Q50", Journal of Shanghai Second Medical College; 1981, 1, 75-86), the simple addition of drug effects is considered to be independent of each other The addition of , its q calculation formula is:
q＝E A/B/(E A+E B-E A·E B), q=E A/B /(E A +E B -E A ·E B ),
其中:E A/B为组合物A/B的实际共用药效(例如抑瘤率),E A和E B分别为组合物A/B的组分A和组分B的单药实际药效,(E A+E B-E A·E B)为组分A和组分B的根据其单药实际药效的理论单纯相加预期效应。q＝1.00反映单纯相加预期作用。 Wherein: EA /B is the actual combined efficacy (such as tumor inhibition rate) of composition A/B, EA and EB are the actual drug efficacy of component A and component B of composition A/ B , respectively , (E A +E B -E A ·E B ) is the theoretical pure additive expected effect of component A and component B according to the actual efficacy of its single drug. q=1.00 reflects purely additive expected effects.
在本发明以下实施例中,根据上述金正均法进行的药物A和B联合给药的共用药效作用判断如下：当药物A和B的组合物的药效无治疗意义(例如r≦30％)时，则组合物未显示出共用作用，亦或者说视为可忽略不计的共同作用。当组合物的药效有治疗意义(例如r＞30％)时，若实际药效/预期药效比q＝1.00则说明组合物的共用药效为单独药效的相加，符合理论单纯相加预期；若实际药效/预期药效比q＞1.00则说明组合物的共用药效存在明显协同作用，即实际作用超过理论单纯相加预期；若实际药效/预期药效比q＜1.00则说明该组合物的共用药效存在明显拮抗作用，即实际作用不及理论单纯相加预期。In the following examples of the present invention, the combined pharmacodynamic effect of the combined administration of drugs A and B according to the above-mentioned Kim Jong-jun method is judged as follows: when the drug efficacy of the combination of drugs A and B has no therapeutic significance (for example, r≦30% ), the composition showed no synergy, or considered negligible synergy. When the drug efficacy of the composition has therapeutic significance (for example, r>30%), if the actual drug efficacy/expected drug efficacy ratio q=1.00, it means that the combined efficacy of the composition is the addition of the individual drug effects, which is consistent with the theoretical simple phase. If the actual efficacy/expected efficacy ratio q>1.00, it means that the combined efficacy of the composition has a significant synergistic effect, that is, the actual effect exceeds the theoretical purely additive expectation; if the actual efficacy/expected efficacy ratio q<1.00 Then it shows that the combined efficacy of the composition has obvious antagonistic effect, that is, the actual effect is not as expected by the simple addition of the theory.
在上述金正均法的q计算基础上,张效文、金正均等进一步使q判断更适合试验实际(张效文等，用Q值估计合并用药效果的新方法:“双30法”,上海第二医学院学报,1985,5,p353-354)。他们以q＝0.85-1.15反映单纯相加预期作用。On the basis of the q calculation of the above-mentioned Jin Zhengjun method, Zhang Xiaowen, Jin Zhengjun and others further made the q judgment more suitable for the actual test (Zhang Xiaowen et al., A new method for estimating the effect of combined medication with Q value: "Double 30 Method", Journal of Shanghai Second Medical College , 1985, 5, p353-354). They reflect purely additive expected effects with q=0.85-1.15.
在本发明以下实施例中,基本上根据上述张效文-金正均法进行联合给药的共用作用判断,即：当组合物A/B组并不显示有治疗意义的药效(例如r≦30％)时,A和B联合给药也就未显示有治疗意义的共用作用,在本发明中视作可忽略不计的共同作用。当组合物组显示有治疗意义的药效(例如r≧30％)时,如果实际/预期比q在0.85与1.15之间则该组合物的共用作用为相加作用(实际作用符合理论单纯相加预期)；如果实际/预期比q>1.15则该组合物的共用作用为明显协同作用(实际作用超理论单纯相加预期)，如果实际/预期比q<0.85则该组合物的共用作用为明显拮抗作用(实际作用不及理论单纯相加预期)。In the following examples of the present invention, based on the above-mentioned Zhang Xiaowen-Jin Zhengjun method, the combined effect of combined administration is judged, that is: when the composition A/B group does not show a therapeutically significant effect (for example, r≦30%) ), the co-administration of A and B also does not show a therapeutically meaningful synergy, which is regarded as a negligible synergy in the present invention. When the composition group showed therapeutically meaningful efficacy (eg, r≧30%), the combined effect of the composition was additive if the actual/expected ratio q was between 0.85 and 1.15 (the actual effect was in line with the theoretical pure phase) If the actual/expected ratio q>1.15, the synergistic effect of the composition is obviously synergistic (the actual effect exceeds the theoretical purely additive expectation), and if the actual/expected ratio q<0.85, the synergistic effect of the composition is Obvious antagonistic effect (the actual effect is not as expected by the theoretical addition).
实施例1：组合物的制备Example 1: Preparation of the composition
按照上述本发明的药物组合物的制备方法，可以配制出本发明众多不同的局部药物组合物。以下列出本发明的药物组合物的制备试验的几个例子。According to the above-mentioned preparation method of the pharmaceutical composition of the present invention, many different topical pharmaceutical compositions of the present invention can be formulated. Several examples of preparation tests of the pharmaceutical composition of the present invention are listed below.
1、液体注射剂的制备1. Preparation of liquid injection
按所需浓度量取药物组合物中的必要组成(例如5g人免疫球蛋白、13g甘氨酸和10g乙酸)、任选存在的其他组分及以及定容至总体积(例如100ml)的液体载体(例如注射用水)，并将它们缓慢混合均匀，除菌过滤后分装为所需量(例如10ml/瓶)储存备用。该制备物(例如5％人免疫球蛋白/13％甘氨酸/10％乙酸水溶液)可作为液体药物局部给药。The necessary components (eg, 5 g of human immunoglobulin, 13 g of glycine, and 10 g of acetic acid), optional other components, and a liquid carrier (eg, 100 ml) of the pharmaceutical composition are dosed to the desired concentration in the pharmaceutical composition. For example, water for injection), and slowly mix them evenly, after sterilization and filtration, divide them into the required amount (for example, 10ml/bottle) and store them for later use. The preparation (eg, 5% human immunoglobulin/13% glycine/10% acetic acid in water) can be administered topically as a liquid drug.
2、注射用粉针剂的制备2. Preparation of powder for injection
按所需浓度量取药物组合物中的必要组成(例如5g人免疫球蛋白、1g亚甲蓝)、任选存 在的其他组分及以及定容至总体积(例如100ml)的液体载体(例如注射用水)，并将它们缓慢混合均匀，除菌过滤后分装为所需量(例如10ml/瓶)进行冷冻干燥及压塞、轧盖，制备为除菌干粉备用。The necessary components (eg, 5 g of human immunoglobulin, 1 g of methylene blue), optional other components, and a liquid carrier (eg, 100 ml) of the pharmaceutical composition are dosed to the desired concentration in the pharmaceutical composition Water for injection), slowly mix them evenly, and after sterilization filtration, pack into the required amount (for example, 10ml/bottle), freeze-dry, cork, and cap, and prepare as sterilized dry powder for later use.
按各组分所需浓度将所需量的除菌干粉(例如1瓶上述干粉)复溶于所需量的除菌液体(例如注射用水)为所需复溶液(例如5％人免疫球蛋白/1％亚甲蓝水溶液)后即可作为液体药物局部给药。According to the required concentration of each component, the required amount of sterile dry powder (for example, 1 bottle of the above dry powder) is reconstituted in the required amount of sterile liquid (for example, water for injection) to obtain the desired reconstituted solution (for example, 5% human immunoglobulin). /1% methylene blue aqueous solution) can be administered locally as a liquid drug.
3、外用液剂的制备3. Preparation of external solution
按所需浓度量取药物组合物中的必要组成(例如5g人免疫球蛋白、13g甘氨酸和10g乙酸)、任选存在的其他组分及以及定容至总体积(例如100ml)的液体载体(例如注射用水)，并将它们缓慢混合均匀。该制备物(例如5％人免疫球蛋白/13％甘氨酸/10％乙酸水溶液)可直接用作外用液剂的液体药物局部给药。The necessary components (eg, 5 g of human immunoglobulin, 13 g of glycine, and 10 g of acetic acid), optional other components, and a liquid carrier (eg, 100 ml) of the pharmaceutical composition are dosed to the desired concentration in the pharmaceutical composition. such as water for injection) and slowly mix them well. The preparation (eg, 5% human immunoglobulin/13% glycine/10% acetic acid in water) can be directly used as a topical solution for topical administration of liquid medicines.
4、雾化剂的制备4. Preparation of Atomizer
按所需浓度(如表1所述)量取药物组合物中的必要组成(例如3g或5g人免疫球蛋白和13g甘氨酸和10g乙酸)、和以下雾化剂辅料：甘油(2.5g)、聚山梨酯-80(1.5g)、苯扎氯铵(0.02g)、微晶纤维素-羧甲基纤维素钠(1.5g)，然后加入定容至总体积(例如100ml)的溶媒(例如注射用水)，并将它们缓慢混合均匀备用。该制备物可作为喷雾剂原液，加入喷雾器后可直接喷洒在靶区形成液体药物。Measure the necessary components in the pharmaceutical composition (eg 3g or 5g human immunoglobulin and 13g glycine and 10g acetic acid) in the desired concentration (as described in Table 1), and the following aerosol excipients: glycerol (2.5g), Polysorbate-80 (1.5 g), benzalkonium chloride (0.02 g), microcrystalline cellulose-sodium carboxymethyl cellulose (1.5 g), then add a vehicle (e.g. water for injection), and slowly mix them well and set aside. The preparation can be used as a spray stock solution, and can be directly sprayed on the target area to form a liquid medicine after being added to the sprayer.
实施例2：Ig局部活性的发现及其组合物优选Example 2: Discovery of Ig Topical Activity and Its Composition Preferences
现有技术中的免疫球蛋白，其在制备肿瘤治疗药中的应用完全是基于其常规活性(例如免疫调节药理)，应用范围狭小，主要用作辅助治疗药物而非治疗药物。免疫球蛋白新应用的开发，实质上是免疫球蛋白新药理的开发。The application of immunoglobulins in the prior art in the preparation of tumor therapeutic drugs is entirely based on their conventional activities (eg, immunomodulatory pharmacology), and has a narrow application range, and is mainly used as an adjuvant therapeutic drug rather than a therapeutic drug. The development of new applications of immunoglobulin is essentially the development of new pharmacology of immunoglobulin.
在一个试验中，试验动物为BALB/c小鼠，建模细胞为乳腺癌4T1细胞，以100ul(每毫升1×10 6个细胞)/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(瘤体平均体积317mm 3)随机分为2个阴性对照组(01、02)和12个药物研究组(1-12)。阴性对照物为生理盐水，研究药物如下表所示,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。药物均为水溶液，按实施例1的制备方法配置而成。各实验组分别以下表所示的全身性给药(静脉注射)、靶区局部给药(瘤内注射)、和混合给药(静脉注射+瘤内注射)方式用药。各组均每3日用药一次，一共3次，注射量150μl/只/次。用药结束后3日，对动物进行安乐死，解剖后测定瘤重，并从阴性对照组计算抑瘤率，结果示于表2。 In one experiment, the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1×10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model. Successfully modeled experimental animals (average tumor volume 317 mm 3 ) were randomly divided into 2 negative control groups (01, 02) and 12 drug study groups (1-12). The negative control is normal saline, the research drugs are shown in the table below, and the human immunoglobulins are the freeze-dried intravenous human immunoglobulins in Table 1. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1. Each experimental group was administered by systemic administration (intravenous injection), local administration in the target area (intratumoral injection), and mixed administration (intravenous injection + intratumoral injection) as shown in the table below. Each group was administered once every 3 days, a total of 3 times, and the injection volume was 150 μl/only/time. The animals were euthanized 3 days after the drug administration, the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 2.
表2Table 2
Figure PCTCN2022076823-appb-000002
Figure PCTCN2022076823-appb-000002
Figure PCTCN2022076823-appb-000003
Figure PCTCN2022076823-appb-000003
通常认为,同一组合物在B技术方案中的抗肿瘤药效超过A方案中的药效动力学提高预期极限(一般为<150％,或从无作用变为有效作用,或瘤重差异有统计学意义),就很可能是不同药理的药物效应。It is generally believed that the anti-tumor efficacy of the same composition in technical scheme B exceeds the expected limit of pharmacodynamic improvement in scheme A (generally <150%, or from no effect to effective effect, or statistical differences in tumor weight). scientific significance), it is likely to be the drug effect of different pharmacology.
在上表中，细胞毒药物(5-氟尿嘧啶)的静脉注射组(组4)和瘤内注射组(组9)显示出几无区别的抑瘤率,尽管有人认为:瘤体内用药可提高其局部药物浓度从而会明显提高其药效。该结果说明该药瘤内注射时并未从本质上改变其常规活性,即靶向肿瘤细胞抑制其增殖。因而，除非置之于缓释***中，常规抗肿瘤药迄今仍然主要是全身性给药而非耐受性较差而药效提高并不明显的局部给药。In the above table, the intravenous injection group (group 4) and the intratumoral injection group (group 9) of the cytotoxic drug (5-fluorouracil) showed almost indistinguishable tumor inhibition rates, although it has been suggested that intratumoral administration may improve the The local drug concentration will thus significantly increase its efficacy. The results indicated that the drug did not substantially change its conventional activity during intratumoral injection, that is, targeting tumor cells to inhibit their proliferation. Therefore, unless it is placed in a sustained-release system, conventional antitumor drugs are still mainly systemically administered so far rather than local administration with poor tolerance and little improvement in efficacy.
在上表中，化学消融剂(乙醇)的静脉注射组(组4)和瘤内注射组(组9)显示出重大的抑瘤率差异(>150％),合乎其在两种给药方案中有不同药理反应的预期。全身给药乙醇与饮酒的药理一致,而局部给药乙醇的药理为局部化学作用可致病变组织破坏。In the table above, the intravenous injection group (group 4) and the intratumoral injection group (group 9) of the chemoablation agent (ethanol) showed a significant difference (>150%) in tumor inhibition rate, in line with its two dosing regimens Different pharmacological responses are expected. The pharmacology of systemic administration of ethanol is consistent with that of drinking alcohol, while the pharmacology of local administration of ethanol is that local chemical action can cause damage to diseased tissue.
通常认为，免疫球蛋白通过靶向免疫***分子实现其作为免疫调节剂的药理，该靶向性决定了其在临床上作为一种以常规给药(例如静脉注射、腔内注射、皮下给药、粘膜给药、肌肉注射、等等)为主的抗肿瘤辅助药物。在上表中，常规用药组1的抑瘤率在可忽略不计的低水平,说明该免疫球蛋白的任何常规活性均不能产生瘤体生长抑制作用。局部给药组6的结果与常规用药组1一致,说明该免疫球蛋白的任何常规活性的局部应用、或其单药局部活性也不能产生瘤体生长抑制作用。这些结果与临床经验一致:对于与本试验中的普通动物一致的患者(病程尚未发展到免疫功能明显下降的普通荷瘤患者)而言,临床上并不推荐提供免疫球蛋白。按照这些结果和现有技术的信息,这种免疫球蛋白不能被直接用作瘤体抑制剂,如果被用作局部活性组分则更是不可思议。It is generally believed that immunoglobulins achieve their pharmacology as immunomodulators by targeting immune system molecules, and this targeting determines their clinical use as a form of routine administration (e.g. intravenous, intracavitary, subcutaneous administration) , mucosal administration, intramuscular injection, etc.)-based anti-tumor adjuvant drugs. In the above table, the tumor inhibition rate of routine drug group 1 was negligibly low, indicating that any routine activity of this immunoglobulin could not produce tumor growth inhibition. The results of topical administration group 6 were consistent with routine administration group 1, indicating that the local application of any conventional activity of the immunoglobulin, or its single-agent local activity, could not produce tumor growth inhibition. These results are consistent with clinical experience: Immune globulin is not clinically recommended for patients consistent with the normal animals in this trial (normal tumor-bearing patients whose disease course has not progressed to a markedly reduced immune function). According to these results and information from the prior art, this immunoglobulin cannot be used directly as a tumor suppressor, and it is even more inconceivable if it is used as a topical active component.
在上表中,常规用药组3的抑瘤率在可忽略不计的低水平,似乎也进一步支持该免疫球 蛋白的组合物也不能产生瘤体生长抑制作用。实际上，加入氨基酸作为免疫球蛋白制品的保护剂是一个非常普遍的应用。按照主流的药学观点,免疫球蛋白(X)仅有常规活性,它与任一药物(Y)的组合物(X/Y)在全身性用药时只能提供这些常规活性去进行联合作用(例如X免疫调节作用与Y常规抗肿瘤作用的组合、或/和X常规抗肿瘤作用与Y常规抗肿瘤作用之间的组合),在局部用药时也只能提供这些常规活性去进行联合作用(例如X免疫调节作用与Y常规抗肿瘤作用或局部作用的组合)。In the above table, the tumor inhibition rate of conventional drug group 3 is at a negligibly low level, which seems to further support that the immunoglobulin composition also cannot produce tumor growth inhibition effect. In fact, the addition of amino acids as protective agents for immunoglobulin preparations is a very common application. According to the mainstream pharmaceutical point of view, immunoglobulin (X) has only conventional activities, and its combination (X/Y) with any drug (Y) can only provide these conventional activities for combined action when it is administered systemically (for example, The combination of X immunomodulatory effect and Y conventional anti-tumor effect, or/and the combination between X’s conventional anti-tumor effect and Y’s conventional anti-tumor effect), can only provide these conventional activities for combined effect in topical administration (e.g. Combination of X immunomodulatory effect and Y conventional antitumor effect or local effect).
然而,组合物组8与组合物组3比较,瘤重差异有统计学意义(p<0.05),抑瘤率差异高达10倍以上,说明该药物组合物在组8中通过不同于组3的新药理实现了瘤体组织消融。与其相应组成单药组相比较,组合物组8的抑瘤率也超出理论单纯相加效应的预期范围,其实际/预期比(q)大于1.15,显示出上述新药理应为局部协同作用药理。免疫球蛋白在该局部协同作用药理中提供的活性,显然超出其任何常规活性的预期,是一种局部活性。However, compared with the composition group 3, the difference in tumor weight was statistically significant (p<0.05), and the difference in tumor inhibition rate was as high as 10 times or more, indicating that the drug composition in group 8 was different from group 3. The new pharmacology realizes tumor tissue ablation. Compared with its corresponding single drug group, the tumor inhibition rate of composition group 8 also exceeded the expected range of theoretical pure additive effect, and its actual/expected ratio (q) was greater than 1.15, indicating that the above-mentioned new pharmacology should be a local synergistic pharmacology. The activity provided by the immunoglobulin in this local synergistic pharmacology is clearly beyond the expectations of any conventional activity and is a local activity.
此外，免疫球蛋白提供该局部活性(与其它成分共用能形成局部协同作用)的药理条件似乎非常严格。在上表中，研究组11与研究组7之间的瘤重差异无统计学意义(p>0.05)且它们的抑瘤率均未显示有治疗意义，研究组12与研究组6之间的瘤重差异无统计学意义(p>0.05)且它们的抑瘤率均未显示有治疗意义，这2个组合物组均未显示出协同药效。这些结果说明，免疫球蛋白提供上述局部活性的一个必要技术条件是：免疫球蛋白与其协同物只有在靶区局部共用的条件下才能产生协同作用；而本发明组合物的一个组成特征是：必须适于局部给药的组成,例如其中所含载体必须包括液体载体。Furthermore, the pharmacological conditions under which immunoglobulins provide this local activity (combined with other components to form a local synergy) appear to be very stringent. In the above table, the difference in tumor weight between study group 11 and study group 7 was not statistically significant (p>0.05) and neither of their tumor inhibition rates showed therapeutic significance. There was no significant difference in tumor weight (p>0.05), and their tumor inhibition rates did not show therapeutic significance, and neither of the two composition groups showed synergistic efficacy. These results indicate that a necessary technical condition for immunoglobulin to provide the above-mentioned local activity is: immunoglobulin and its synergist can only produce synergy under the condition of local sharing of the target area; and one of the compositional features of the composition of the present invention is that it must be Compositions suitable for topical administration, eg, the carrier contained therein, must include a liquid carrier.
通常认为，药靶是药理因而也是药效的关健之一。对于上述实验中包含免疫球蛋白的组合物在不同用药中显示出的完全不同药效，很可能是因为它们的药靶、药理完全不同。常规组合物通过常规用药进入体内，主要是分布在血液中并以血药的形式去靶向(例如研究组3)，靶可以是免疫***、癌细胞、等等。而局部组合物通过局部给药进入体内，主要是依靠短暂外力在局部病灶(肿瘤组织)内的间隙中分布和渗透(例如研究组8)，其所作用的首先是组织、然后才是其中的内容物(癌细胞、免疫细胞)。上述研究组8比组3高出的37倍以上的抑瘤率显示出的是异乎其常的类化学消融结果，该结果显然来自异乎其常的局部协同药理，且看上去该协同药理与常规活性例如免疫调节药理、癌细胞抑制药理均无明显关系。It is generally believed that the drug target is one of the keys to the pharmacology and therefore the efficacy of the drug. The completely different pharmacological effects of the immunoglobulin-containing compositions in the above experiments are likely to be due to their completely different drug targets and pharmacology. Conventional compositions enter the body through conventional drugs, mainly distributed in the blood and de-targeted in the form of blood drugs (eg study group 3), the target can be the immune system, cancer cells, and so on. The topical composition enters the body through local administration, mainly relying on short-term external force to distribute and penetrate in the gap in the local lesion (tumor tissue) (for example, research group 8), which acts on the tissue first, and then in it. Contents (cancer cells, immune cells). The more than 37-fold higher tumor inhibition rate in the above study group 8 than in group 3 shows an unusual chemical-like ablation result, which is clearly derived from an unusual local synergistic pharmacology, and it seems that the synergistic pharmacology is consistent with conventional methods. Activities such as immunomodulatory pharmacology, cancer cell inhibitory pharmacology were not significantly related.
众所周知,药学并非化学,药物并非完全由化学定义的物质,而是构-效统一的活性物质。传统观念中,提供相同活性(同效,例如细胞毒作用)的不同物质(不同构,例如上述实验中的亚甲蓝类染料和常规抗肿瘤药)很容易被视作不同药物。然而,提供不同活性(不同效,例如免疫调节作用、常规抗肿瘤作用和局部协同作用)的相同化学物质(同构,例如免疫球蛋白)则很容易被偏见视作相同药物。As we all know, pharmacy is not chemistry, and medicine is not a substance completely defined by chemistry, but an active substance with unified structure and effect. In the traditional concept, different substances (different structures, such as methylene blue dyes and conventional antineoplastic drugs in the above experiments) that provide the same activity (same effect, such as cytotoxicity) can easily be regarded as different drugs. However, identical chemicals (isomorphisms, such as immunoglobulins) that provide different activities (different effects, such as immunomodulatory effects, conventional antitumor effects, and local synergism) can easily be prejudiced as the same drug.
在本发明中,免疫球蛋白的免疫调节作用被记作A1、该作用所需的特征组成被记作X1；其常规抗肿瘤作用(假如有的话)被记作A2、该作用所需的特征组成被记作X2；其局部活性(有效局部协同作用)被记作A3、该活性所需的特征组成被记作X3(例如上述及以下实施例所述的组成特征)。从药学而非化学的角度,这是可提供不同活性(A1、A2、A3)、也具有不 同给药组成特征(X1、X2、X3)的不同构效的三种免疫球蛋白(A1X1、A2X2、A3X3)。In the present invention, the immunomodulatory action of immunoglobulin is denoted as A1, the characteristic composition required for this action is denoted as X1; its conventional antitumor action (if any) is denoted as A2, and the required characteristic composition for this action is denoted as A2. The characteristic composition is denoted X2; its local activity (effective local synergy) is denoted A3, and the characteristic composition required for this activity is denoted X3 (such as those described in the above and following examples). From a pharmaceutical rather than chemical point of view, these are three immunoglobulins (A1X1, A2X2) that provide different activities (A1, A2, A3) and also have different structure-effect profiles (X1, X2, X3) , A3X3).
众所周知,假如存在明显、且不说有效的药效的话,A1X1和A2X2应当在以上实验的全身性用药动物中显示出来。在本实施例和以下实施例还证实,应用A3X3可以制备出远远超过A1X1和A2X2的预期技术效果(给药效率)的本发明组合物。It is well known that A1X1 and A2X2 should be shown in systemically administered animals in the above experiments if there is a significant, not to say, effective drug effect. In this example and the following examples, it is also confirmed that the application of A3X3 can prepare the composition of the present invention which far exceeds the expected technical effect (dosing efficiency) of A1X1 and A2X2.
以上研究以及其它类似研究说明,本发明的组合物的功能组合或组合物,并非1):免疫球蛋白的免疫作用和共用物局部作用(B1)的组合(A1/B1),或A1X1/B1Y组合物,也非2):免疫球蛋白的常规抗肿瘤作用(假如有的话)(A2)和共用物局部作用(B1)的组合(A2/B1),或A2X2/B1Y组合物,而是3):免疫球蛋白的局部活性(局部协同作用,A3)和共用物对免疫球蛋白的局部协同作用(B2)的组合(A3/B2),或A3X3/B2Y组合物。The above studies, and other similar studies, demonstrate that the functional combination or composition of the composition of the present invention is not 1): a combination of immunoglobulin immune effect and co-use local effect (B1) (A1/B1), or A1X1/B1Y Composition, nor 2): a combination (A2/B1) of the conventional antitumor effect of immunoglobulin (if any) (A2) and the local effect of the concomitant (B1), or an A2X2/B1Y composition, but 3): Local activity of immunoglobulin (local synergy, A3) and combination of local synergy (B2) of immunoglobulin (A3/B2), or A3X3/B2Y composition.
实施例3：Ig局部活性提供的短期协同作用药理研究Example 3: Pharmacological Study of Short-Term Synergy Provided by Ig Local Activity
在一个试验中，试验动物为裸小鼠，建模细胞为肉瘤S180，以100ul(每毫升1×10 6个细胞)/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(荷肉瘤小鼠，瘤体平均体积226mm 3)随机分为8个试验组(1个阴性对照组和7个研究组)。阴性对照物为生理盐水，研究药物如下表所示,其中的人免疫球蛋白为表1中的重组人IgG。药物均为水溶液，按实施例1的制备方法配置而成。各组均每3日用药一次，一共3次，行瘤内注射，注射量125μl/只/次。用药结束后3日，对动物进行安乐死，解剖后测定瘤重，并从阴性对照组计算抑瘤率，结果示于表3。 In one experiment, the experimental animals were nude mice, the model cells were sarcoma S180, and 100ul (1×10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animals. Successfully modeled experimental animals (sarcoma-bearing mice, with an average tumor volume of 226 mm 3 ) were randomly divided into 8 experimental groups (1 negative control group and 7 research groups). The negative control is normal saline, the research drugs are shown in the following table, and the human immunoglobulin is the recombinant human IgG in Table 1. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1. Each group was administered once every 3 days for a total of 3 times. The animals were euthanized 3 days after the drug administration, and the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 3.
表3table 3
Figure PCTCN2022076823-appb-000004
Figure PCTCN2022076823-appb-000004
裸小鼠为先天性无胸腺的裸(nude)小鼠的位于第11对染色体上的隐性突变基因“nu”导入BALB/c小鼠所获小鼠。其胸腺仅有残迹或异常上皮，不能使T细胞正常分化，缺乏成熟T细胞的辅助、抑制和杀伤功能，免疫力低下。其对异种生物物质有高度的免疫耐受性,例如对异种癌细胞几无移植免疫反应。在现有技术中，荷瘤裸小鼠被广泛用作化疗模型而非免疫治疗模型。按照现有技术的教导，免疫球蛋白(既使是异种免疫球蛋白)作为免疫调节药物不应在荷瘤裸小鼠实验中显示出免疫抑瘤作用、更不用说局部作用、更不要说局 部化学作用。在上表中，研究组1的结果符合该预期。Nude mice are congenital athymic nude mice obtained by introducing a recessive mutation gene "nu" on the 11th pair of chromosomes into BALB/c mice. The thymus has only remnants or abnormal epithelium, which cannot make T cells differentiate normally, lacks the auxiliary, inhibitory and killing functions of mature T cells, and has low immunity. It has a high degree of immune tolerance to xenogeneic biological substances, for example, there is almost no transplant immune response to xenogeneic cancer cells. In the prior art, tumor-bearing nude mice are widely used as models of chemotherapy rather than immunotherapy. As taught in the prior art, immunoglobulins (even xenogeneic immunoglobulins) as immunomodulatory drugs should not exhibit immunosuppressive, let alone local effects, let alone local effects in tumor-bearing nude mice experiments chemical action. In the table above, the results of Study Group 1 met this expectation.
通常认为，局部共用化学作用必须是2个具有化疗作用的单药的共用作用。出人意料的是，组合物组3、5、7与组1之间的差异有统计学意义(P<0.05),且它们各自的实际/预期比(q)均大于1.15,均显示出短期协同作用。该短期协同作用理应包括局部化学协同作用。It is generally believed that the topical shared chemical action must be the shared action of 2 single agents with chemotherapeutic effect. Unexpectedly, the differences between composition groups 3, 5, 7 and group 1 were statistically significant (P<0.05), and their respective actual/expected ratios (q) were all greater than 1.15, all showing short-term synergy. . This short-term synergy should include local chemical synergy.
实施例4：Ig局部活性提供的中长期协同作用药理研究Example 4: Pharmacological study of mid- and long-term synergy provided by Ig topical activity
以裸小鼠为实验动物，以人肝癌HepG2细胞为移植瘤建模细胞,按公知方法建模,在每只动物右侧腋部皮下注射1×10 5个癌细胞。移植瘤体生长至平均体积127mm 3时将荷瘤动物随机分为3个系列A、B、C,且每个系列分别被随机分为5组(组别0、1、2、3、4),其中各系列中：组0均为阴性对照组，给予生理盐水；组1均为化学消融对照组，给予无水乙醇；组2均为共用物组，给予20％甘氨酸/10％乙酸；组3均为免疫球蛋白组，给予4％人免疫球蛋白；组4均为组合物组，给予4％人免疫球蛋白/20％甘氨酸/10％乙酸。药物均为水溶液，研究药物按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的人IgG。 Nude mice were used as experimental animals, and human hepatoma HepG2 cells were used as transplanted tumor model cells. According to the well-known method, 1×10 5 cancer cells were subcutaneously injected into the right armpit of each animal. When the transplanted tumor grew to an average volume of 127 mm, the tumor-bearing animals were randomly divided into 3 series A, B, and C, and each series was randomly divided into 5 groups (group 0, 1, 2, 3, and 4). , in each series: group 0 is a negative control group, given normal saline; group 1 is a chemical ablation control group, given anhydrous ethanol; group 2 is a shared substance group, given 20% glycine/10% acetic acid; Group 3 is an immunoglobulin group, given 4% human immunoglobulin; group 4 is a composition group, given 4% human immunoglobulin/20% glycine/10% acetic acid. The medicines are all aqueous solutions, and the research medicines are prepared according to the preparation method of Example 1, and the human immunoglobulin in it is the human IgG in Table 1.
动物每2日用药一次，一共3次,系列A各组行瘤内注射(75μl/只/次)，系列C各组行尾静脉注射(75μl/只/次),系列B各组行瘤内注射(75μl/只/次)和尾静脉注射(75μl/只/次),其中组别4瘤内注射20％甘氨酸/10％乙酸和尾静脉注射4％人免疫球蛋白。第一次用药第7日和第21日，分别测定各系列各组别动物的瘤体体积，并相对于各系列阴性对照组分别计算1-4组的7日体积抑瘤率(r7)和21日体积抑瘤率(r21)，结果示于下表4。Animals were administered once every 2 days for a total of 3 times. Each group in series A received intratumoral injection (75 μl/animal/time), each group in series C received tail vein injection (75 μl/animal/time), and each group in series B received intratumoral injection Injection (75μl/animal/time) and tail vein injection (75μl/animal/time), including group 4 intratumoral injection of 20% glycine/10% acetic acid and tail vein injection of 4% human immunoglobulin. On the 7th and 21st days of the first medication, the tumor volume of animals in each series and each group was measured, and the 7-day volume tumor inhibition rate (r7) and The volume tumor inhibition rate (r21) on the 21st day, the results are shown in Table 4 below.
表4各系列不同药物组别对应的不同期体积抑瘤率(r 7、r 21)结果 Table 4 Results of volume tumor inhibition rate (r 7 , r 21 ) in different stages corresponding to different drug groups in each series
Figure PCTCN2022076823-appb-000005
Figure PCTCN2022076823-appb-000005
在上表中，无水乙醇(组1)和甘氨酸/乙酸(组2)均在现有技术中显示一定的细胞毒活性,但它们在系列C中的r 7和r 21均为<5％,说明它们可用作常规活性的任何活性(包括该细胞毒活性)均无短期和中长期抑瘤作用；它们的局部活性在系列A中的短期药效(r 7)远远高于中长期药效(r 21),合乎现有技术中“局部活性均不可持续”的预期；而它们在系列B的药效与系列A相似,说明其可用作常规活性的任何活性(包括细胞毒活性)并未增强其局部活性。免疫球蛋白(组3)在系列A、B、C中的中长期药效也如同短期药效(r 7和r 21均为<5％),说明其任 何可用作常规活性或局部活性的活性(包括免疫调节活性)、以及它们的联合使用均无短期和中长期抑瘤作用。总之,免疫球蛋白与其共用组分单用时,它们各自可用作常规活性或局部活性的任何活性、以及这些常规活性和局部活性的联合作用均未显示抑瘤的中长期作用。 In the above table, both absolute ethanol (group 1) and glycine/acetic acid (group 2) show some cytotoxic activity in the prior art, but their r 7 and r 21 in series C are both <5% , indicating that any activity (including this cytotoxic activity) that they can be used as conventional activity has no short-term and medium- and long-term tumor inhibitory effects; the short-term efficacy (r 7 ) of their local activity in series A is much higher than that in the medium and long term Pharmacodynamics (r 21 ), in line with the prior art expectation that "local activity is unsustainable"; and their pharmacodynamics in series B are similar to series A, indicating that they can be used as any activity of conventional activity (including cytotoxic activity ) did not enhance its local activity. The mid- and long-term efficacy of immunoglobulins (group 3) in series A, B, and C was also similar to the short-term efficacy (<5% for both r 7 and r 21 ), indicating that any of its available for routine or topical activity. Activity (including immunomodulatory activity), and their combined use have no short-term or long-term tumor inhibitory effect. In conclusion, when the immunoglobulins and their shared components are used alone, any of the activities that each of them can be used as conventional or topical activities, and the combined effects of these conventional and topical activities do not show mid- and long-term tumor-suppressive effects.
在上表中，免疫球蛋白组合物(组4)在系列C中的r 7和r 21均为<5％,说明组合物各组分可用作常规活性的任何活性(包括该细胞毒活性)的联合使用均无短期和中长期抑瘤作用；在系列B中其药效与共用组分(组2)相似,说明免疫球蛋白可用作常规活性的任何活性(包括免疫调节活性)与共用组分局部活性的联合使用并未超过该局部活性的短期活性预期(q 7>1.00)和中长期活性预期(q 21>1.00)。总之,免疫球蛋白可用作常规活性的任何活性均未显示对其共用组分的任何常规活性或局部活性增强的中长期作用。 In the table above, the immunoglobulin composition (group 4) has both r7 and r21 in series C < 5 %, indicating that the components of the composition can be used as any activity for conventional activities (including this cytotoxic activity) ) had no short-term and mid-term and long-term tumor inhibitory effects; in series B, its efficacy was similar to that of the common component (group 2), indicating that any activity (including immunomodulatory activity) that immunoglobulins can be used as conventional activities is comparable to The combined use of the topical activity of the common components did not exceed the short-term activity expectations (q 7 >1.00) and medium and long-term activity expectations (q 21 >1.00) for this topical activity. In conclusion, none of the activities for which the immunoglobulins can be used as a conventional activity have shown any conventional or locally enhanced mid- to long-term effects on their shared components.
然而,在系列A中,通过组2-4的r 7计算出的Ig组合物的q 7>1.15,说明Ig局部活性提供了如以上实施例2中所述的超过局部加和作用预期的短期协同作用,该短期协同作用是免疫球蛋白组合物的主要短效药理；通过组2-4的r 21计算出的q 21>q 7>1.15,说明Ig局部活性提供了协同性(或超预期性)超过短期协同作用的中长期协同作用。该中长期协同作用很可能主要是Ig局部活性与其共用组分局部活性的联合使用的次生协同作用,也是免疫球蛋白组合物的主要中长效药理。这些结果还说明,免疫球蛋白与其它成分共用能形成中长期协同作用的药理条件似乎非常严格：免疫球蛋白只有在可提供局部活性的的条件下才能与其短期协同物产生该中长期协同作用。 However, in Series A , the q7 of the Ig composition calculated by the r7 of groups 2-4 was >1.15, indicating that the Ig local activity provided a short - term excess of the local additive effect expected as described in Example 2 above. Synergy, this short-term synergy is the main short-acting pharmacology of immunoglobulin compositions; q 21 >q 7 >1.15 calculated from r 21 of groups 2-4, indicating that the local activity of Ig provides synergy (or more than expected) sex) mid- and long-term synergies over short-term synergies. This mid- and long-term synergy is probably mainly the secondary synergy of the combined use of the local activity of Ig and the local activities of its shared components, and is also the main mid- and long-term pharmacology of the immunoglobulin composition. These results also suggest that the pharmacological conditions under which immunoglobulins and other components can form mid- and long-term synergy appear to be very stringent: immunoglobulins can only produce this mid- and long-term synergy with their short-term synergists under conditions that provide local activity.
以下实施例5和6进一步研究在局部组织反应器中可以与上述Ig局部活性发生有治疗意义的药理反应的反应物。Examples 5 and 6 below further investigate reactants that can have a therapeutically meaningful pharmacological response to the above-described local activity of Ig in a local tissue reactor.
实施例5：Ig局部活性的反应物研究：泛实体肿瘤Example 5: Reactant Studies for Local Activity of Ig: Pan-Solid Tumors
以裸小鼠为实验动物，分为如下表所示的4个研究系列(A、B、C、D),系列A、B、C、D分别以下列癌细胞进行移植瘤建模：人胰腺癌Panc-1细胞、人结肠癌Lovo细胞、人肝癌HepG2细胞、人胃癌BGC823细胞。建模按公知方法进行,每只动物右侧腋部皮下注射1×10 5个癌细胞,各系列分别生长至如下平均体积时为建模成功:109±21mm 3、118±18mm 3、106±23mm 3、114±27mm 3。成功建模的每个系列分别被随机分为如下表所示的5组(组0、1、2、3、4),其中：组0均为阴性对照组，给予生理盐水；组1均为化学消融对照物组，给予无水乙醇；组2均为共用物组，给予2％亚甲蓝；组3均为免疫球蛋白组，给予4％人免疫球蛋白；组4均为组合物组，给予4％人免疫球蛋白/2％亚甲蓝。药物均为水溶液，研究药物按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。各系列各组动物均每2日用药一次，一共3次，行瘤内注射，注射量75μl/只。给药时观察实验动物的药物局部刺激强度(例如挣扎强度)，分别以生理盐水和无水乙醇的局部刺激现象为1和5进行强度估算。第一次用药第7日和第21日，分别测定各系列各组动物的瘤体体积，并相对于各系列阴性对照组分别计算1-4组的7日体积抑瘤率(r7)和21日体积抑瘤率(r21)，结果示于下表5中。 Nude mice were used as experimental animals and were divided into 4 research series (A, B, C, D) as shown in the table below. Series A, B, C, and D were modeled with the following cancer cells: human pancreas. Cancer Panc-1 cells, human colon cancer Lovo cells, human liver cancer HepG2 cells, human gastric cancer BGC823 cells. The modeling was carried out according to the known method. 1×10 5 cancer cells were subcutaneously injected into the right axilla of each animal, and the modeling was successful when each series grew to the following average volumes: 109±21 mm 3 , 118±18 mm 3 , 106± 23mm 3 , 114±27mm 3 . Each series that was successfully modeled was randomly divided into 5 groups (groups 0, 1, 2, 3, 4) as shown in the table below, where: group 0 was a negative control group and was given normal saline; group 1 was a negative control group. The chemical ablation control group was given anhydrous ethanol; the group 2 was the common substance group, which was given 2% methylene blue; the group 3 was the immunoglobulin group, which was given 4% human immunoglobulin; the group 4 was the composition group , given 4% human immunoglobulin/2% methylene blue. The medicines are all aqueous solutions, and the research medicines are prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Animals in each series and each group were given medication once every 2 days for a total of 3 times, and were injected intratumorally with an injection volume of 75 μl per animal. The local irritation intensity (such as struggling intensity) of the experimental animals was observed during administration, and the intensity was estimated by taking the local irritation phenomena of normal saline and absolute ethanol as 1 and 5, respectively. On the 7th and 21st days of the first medication, the tumor volume of each series of animals in each group was measured, and the 7-day volume tumor inhibition rate (r7) and 21 of the 1-4 groups were calculated relative to the negative control group of each series. Daily volume tumor inhibition rate (r21), the results are shown in Table 5 below.
表5各系列不同药物组别对应的体积抑瘤率(r 7、r 21)结果 Table 5 Results of volume tumor inhibition rate (r 7 , r 21 ) corresponding to different drug groups in each series
Figure PCTCN2022076823-appb-000006
Figure PCTCN2022076823-appb-000006
在上表各系列中,通过组2-4的r7和r21计算出的人免疫球蛋白/亚甲蓝组合物的q7和q21均为>1.15,说明Ig局部活性提供了如以上实施例2-4中所述的超过局部加和作用预期的短期协同作用和中长期协同作用。In each series of the above table, the q7 and q21 of the human immunoglobulin/methylene blue composition calculated by the r7 and r21 of groups 2-4 are both >1.15, indicating that the local activity of Ig provides as above Example 2- Short-term synergy and mid- to long-term synergy beyond the expected local additive effect as described in 4.
在上表各系列中,相同药物药效的不同系列差别甚至在同系列不同实验可以接受的范围内(偏差≦20％)。具体而言，组别1均显示出非常接近的短期药效(73.6％≦r7≦89.4％)和中长期药效(21.1％≦r21≦25.8％)，合乎无水乙醇化学消融的非肿瘤细胞特异性预期；组别2也显示出较为接近的短期药效(38.2％≦r7≦47.9％)和中长期药效(20.3％≦r21≦29.2％)，合乎亚甲蓝局部作用的非肿瘤细胞特异性预期；组别3均未显示有冶疗意义的短期药效和中长期药效，其作用未见肿瘤细胞特异性；组别4也都显示出较为接近的短期药效(81.3％≦r7≦91.4％)和中长期药效(70.7％≦r21≦80.1％)。In each series in the table above, the difference between different series of efficacy of the same drug is even within the acceptable range of different experiments in the same series (deviation≤20%). Specifically, group 1 showed very similar short-term efficacy (73.6%≦r7≦89.4%) and medium and long-term efficacy (21.1%≦r21≦25.8%), which were in line with non-tumor cells chemically ablated by absolute ethanol. Specificity is expected; group 2 also showed relatively similar short-term efficacy (38.2%≦r7≦47.9%) and medium and long-term efficacy (20.3%≦r21≦29.2%), in line with the local effect of methylene blue on non-tumor cells Specificity is expected; group 3 did not show short-term efficacy and mid-term and long-term efficacy of therapeutic significance, and its effect was not specific to tumor cells; group 4 also showed relatively similar short-term efficacy (81.3%≤ r7≦91.4%) and medium and long-term efficacy (70.7%≦r21≦80.1%).
本实施例中肿瘤细胞和以上实施例中其它肿瘤细胞构成一个细胞本质很宽泛的肿瘤细胞分布谱，本发明的组合物在其中显示出很类似的短期药效和中长期药效，说明Ig局部活性并无明显的肿瘤细胞特异性，可以与几乎任何实体肿瘤组织发生其药理反应，产生类似的短期协同作用和中长期协同作用。The tumor cells in this example and other tumor cells in the above examples constitute a tumor cell distribution spectrum with a very broad cellular nature, and the composition of the present invention shows very similar short-term efficacy and medium and long-term efficacy in it, indicating that Ig local The activity has no obvious tumor cell specificity, and can have its pharmacological response with almost any solid tumor tissue, producing similar short-term synergy and mid- and long-term synergy.
实验中还发现，各系列组别2的局部刺激强度在组别0和1之间，估值约为2.0；各系列组别3的局部刺激强度在组别0和2之间，估值约为1.0；各系列组别4的局部刺激强度在组别2和3之间，估值约为1.2。药物联合使用产生的毒理作用(或副作用)具有高度不确定性，可以按照上述药理作用的方法进行判断，即依据以下实际副作用/预期副作用比Q判断进行：Q＝Q1/Q2，其中Q1为实际共用副作用，Q2为Ig及其共用物的实际单用副作用得出的预期相加副作用。通常认为，药物副作用的单纯相加是互相独立的副作用的相加。若Q＝1时说明实际共用副作用符合预期，即显示为副作用相加作用；当Q＜1时说明实际共用副作用不及预期，即显示为副作用之间产生了拮抗作用，或产生了安全性协同；当Q＞1时说明实际共用副作用超预期，即显示为副作用之间产生了协同作用，或产生了安全性拮抗。例如，预期相加副作用至少为Ig(A)及其共用物(B)的实际单用副作用中最大者(Q2＞Q2max(Ig，共用物))，于是当Ig(A)及其共用物(B)的实际共用副作用小于该最 大者(Q1＜Q2max(Ig，共用物))时，Q＝Q1/Q2为＜1，说明Ig及其共用物的共用产生了安全性协同。It was also found in the experiment that the local stimulation intensity of each series of group 2 was between groups 0 and 1, and the estimated value was about 2.0; the local stimulation intensity of each series of group 3 was between groups 0 and 2, and the estimated value was about 2.0. is 1.0; the local stimulus intensity of group 4 in each series is between groups 2 and 3, and is estimated to be about 1.2. The toxicological effects (or side effects) produced by the combined use of drugs are highly uncertain, and can be judged according to the above-mentioned methods of pharmacological effects, that is, according to the following actual side effects/expected side effects ratio Q: Q=Q1/Q2, where Q1 is Actual co-use side effects, Q2 is the expected additive side-effects derived from the actual mono-use side effects of Ig and its co-uses. It is generally believed that the mere addition of drug side effects is the addition of mutually independent side effects. If Q=1, it means that the actual side effects of shared use are in line with expectations, which means that the side effects are additive; when Q<1, it means that the actual side effects of shared use are less than expected, which means that there is an antagonism between the side effects, or a safety synergy; When Q>1, it means that the actual side effects of shared use exceed expectations, that is, it is shown that there is a synergy between the side effects, or a safety antagonism is produced. For example, the expected additive side effect is at least the largest of the actual single-use side effects of Ig(A) and its combination (B) (Q2 > Q2max(Ig, combination)), then when Ig(A) and its combination ( When the actual shared side effect of B) is less than the maximum (Q1<Q2max(Ig, co-use)), Q=Q1/Q2 is <1, indicating that the sharing of Ig and its co-use produces safety synergy.
实施例6：Ig局部活性的反应物研究:含成纤维细胞局部病变Example 6: Reactant studies for local activity of Ig: local lesions with fibroblasts
以裸小鼠为实验动物，分为如下表所示的2个研究系列(A、B),系列A、B分别以小鼠胚胎成纤维细胞3T3(0.5×10 4个细胞/只)和小鼠乳腺癌4T1细胞与小鼠胚胎成纤维细胞3T3的混合物(0.5×10 5个细胞/只，3T3与4T1的细胞数量比为1/10)进行药理反应物(结节)建模。建模按公知方法进行，每只动物右侧腋部皮下注射建模细胞,系列A、B分别选择生长至129±32mm3和134±27mm3的动物用作试验。使用肿瘤瘤体间质比测定的公知方法，测出系列B中动物瘤体的平均间质比为39.7％。 Nude mice were used as experimental animals, and they were divided into 2 research series (A, B) as shown in the table below. Series A and B were made of mouse embryonic fibroblasts 3T3 (0.5×10 4 cells/a) and small fibroblasts, respectively. A mixture of murine breast cancer 4T1 cells and mouse embryonic fibroblasts 3T3 (0.5 x 10 5 cells per mouse, 1/10 cell number ratio of 3T3 to 4T1) was used to model pharmacological reactants (nodules). The modeling was carried out according to the known method. The modeled cells were injected subcutaneously in the right axilla of each animal, and the animals of series A and B were selected to grow to 129±32mm3 and 134±27mm3 for the experiment. Using a well-known method for the determination of tumor-tumor-stromal ratio, the mean stromal ratio of the animals in series B was determined to be 39.7%.
成功建模的系列A、B分别被随机分为如下表所示的5组(组0、1、2、3、4)，其中：组0均为阴性对照组，给予生理盐水；组别1均为化学消融对照物组，给予无水乙醇；组2均为共用物组，给予20％甘氨酸/10％乙酸(pH3.9)；组3均为免疫球蛋白组，给予4％人免疫球蛋白；组4均为组合物组，给予4％人免疫球蛋白/20％甘氨酸/10％乙酸(pH3.9)。药物均为水溶液，测试药物按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。系列A、B各组动物均每2日给予一次，一共3次，行瘤内注射，注射量75μl/只。给药时观察实验动物的药物局部刺激强度(例如挣扎强度)，分别以生理盐水和无水乙醇的局部刺激现象为1和5进行强度估算。第一次用药第7日和第21日，分别测定各系列各组别动物的瘤体体积，并相对于各系列阴性对照组分别计算1-4组的7日体积抑瘤率(r7)和21日体积抑瘤率(r21)，结果示于下表6。The successfully modeled series A and B were randomly divided into 5 groups (groups 0, 1, 2, 3, and 4) as shown in the following table, among which: group 0 was a negative control group, given normal saline; group 1 All were chemical ablation control group, given absolute ethanol; group 2 were all shared substance group, given 20% glycine/10% acetic acid (pH3.9); group 3 were all immunoglobulin groups, given 4% human immune globulin Protein; group 4 is a composition group, administered with 4% human immunoglobulin/20% glycine/10% acetic acid (pH 3.9). The medicines are all aqueous solutions, and the test medicines are prepared according to the preparation method of Example 1, and the human immunoglobulins are the freeze-dried intravenous human immunoglobulins in Table 1. The animals in each group of series A and B were given once every 2 days for a total of 3 times. The local irritation intensity (such as struggling intensity) of the experimental animals was observed during administration, and the intensity was estimated by taking the local irritation phenomena of normal saline and absolute ethanol as 1 and 5, respectively. On the 7th and 21st days of the first medication, the tumor volume of animals in each series and each group was measured, and the 7-day volume tumor inhibition rate (r7) and The volume tumor inhibition rate (r21) on the 21st day, the results are shown in Table 6 below.
表6各系列不同药物组别对应的体积抑瘤率(r 7、r 21)结果 Table 6 Results of volume tumor inhibition rate (r 7 , r 21 ) corresponding to different drug groups in each series
Figure PCTCN2022076823-appb-000007
Figure PCTCN2022076823-appb-000007
在上表中，通过组2-4的r7和r21计算出的Ig组合物的q7和q21均为>1.15，说明Ig局部活性提供了有如以上实施例2-5在荷瘤动物中的所述超过局部加和作用预期的短期协同作用和中长期协同作用。In the above table, the q7 and q21 of the Ig composition calculated by the r7 and r21 of groups 2-4 are both >1.15, indicating that the local activity of Ig provides the same as described above in the tumor-bearing animals of Examples 2-5 Short-term synergy and mid- to long-term synergy beyond expectations of local additive effects.
在上表各系列中，Ig组合物的短期药效(49.6％≦r7≦71.2％)和中长期药效(60.1％≦r21≦61.5％)均相差不大，其药效的不同系列差别甚至在同系列不同实验可以接受的范围内(偏差≦20％)。Among the series in the table above, the short-term efficacy (49.6%≦r7≦71.2%) and the mid- and long-term efficacy (60.1%≦r21≦61.5%) of the Ig composition are not much different. Within the acceptable range of different experiments in the same series (deviation≤20%).
本实施例中包含成纤维细胞以及肿瘤细胞/成纤维细胞的结节构成一个细胞本质很宽泛的细胞分布谱，本发明的组合物在其中显示出很类似的短期药效和中长期药效，说明Ig局部活性并无明显的细胞特异性，可以与几乎任何包含成纤维细胞的病变组织发生其药理反应,产生类似的短期协同作用和中长期协同作用。In this example, the nodules containing fibroblasts and tumor cells/fibroblasts constitute a broad spectrum of cellular distribution in which the composition of the present invention shows very similar short-term efficacy and mid-term and long-term efficacy, It shows that the local activity of Ig has no obvious cell specificity, and can have its pharmacological reaction with almost any diseased tissue containing fibroblasts, producing similar short-term synergy and mid- and long-term synergy.
实验中还发现，各系列组别2的局部刺激强度在组别0和1之间，估值约为3.5；各系列组别3的局部刺激强度在组别0和2之间，估值约为1；各系列组别4的局部刺激强度在组别2和3之间，估值约为2.5。预期相加副作用至少为Ig(A)及其共用物(B)的实际单用副作用中最大者(Q2＞Q2max(Ig，共用物))，于是当Ig(A)及其共用物(B)的实际共用副作用小于该最大者(Q1＜Q2max(Ig，共用物))时，Q＝Q1/Q2为＜1，说明Ig及其共用物的共用产生了安全性协同。In the experiment, it was also found that the local stimulation intensity of each series of group 2 was between groups 0 and 1, and the estimated value was about 3.5; the local stimulation intensity of each series of group 3 was between groups 0 and 2, and the estimated value was about is 1; the local stimulus intensity of group 4 in each series is between groups 2 and 3, estimated to be about 2.5. The expected additive side effect is at least the largest of the actual single-use side effects of Ig(A) and its combination (B) (Q2>Q2max(Ig, combination)), so when Ig(A) and its combination (B) When the actual side effect of sharing is less than the maximum (Q1<Q2max(Ig, sharing substance)), Q=Q1/Q2 is <1, indicating that the sharing of Ig and its sharing substance produces safety synergy.
实施例7：Ig提供局部活性所需条件:局部协同物研究及优选Example 7: Conditions required for Ig to provide topical activity: topical synergistic studies and optimization
在一个试验中，试验动物为BALB/c小鼠，建模细胞为乳腺癌4T1细胞，以100ul(每毫升1×10 6个细胞)/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(瘤体平均体积324mm 3)随机分为1个阴性对照组(01)和7个药物研究组(1-7)。阴性对照物为生理盐水，研究药物如下表所示。药物均为水溶液，按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。各实验组均行瘤内注射，每3日用药一次，一共3次，注射量150μl/只/次。用药结束后3日，对动物进行安乐死，解剖后测定瘤重，并从阴性对照组计算抑瘤率，结果示于表7。 In one experiment, the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1×10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model. Successfully modeled experimental animals (with an average tumor volume of 324 mm 3 ) were randomly divided into 1 negative control group (01) and 7 drug study groups (1-7). The negative control is normal saline, and the research drugs are shown in the table below. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each experimental group received intratumoral injection, once every 3 days, for a total of 3 times, and the injection volume was 150 μl/only/time. Three days after the end of administration, the animals were euthanized, the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 7.
表7Table 7
Figure PCTCN2022076823-appb-000008
Figure PCTCN2022076823-appb-000008
在上表中，单药组3所用5-氟尿嘧啶是一种常规用药和瘤内用药均有效的肿瘤细胞抑制药物；单药组4所用二盐酸奎宁是一种瘤内用药浓度≥3时才能作为化学消融剂应用的常规抗疟疾药物；单药组2所用亚甲蓝是一种浓度≥1时才能作为化学消融剂应用的常规活体染料。单药组2、3、4的抑瘤率分别为13％、53％、58％。然而，在与单药组2、3、4对应的组合物组5、6、7之间，组合物组5的抑瘤率是另外两组的135％以上。In the above table, 5-fluorouracil used in single drug group 3 is an effective tumor cell inhibitory drug for both conventional and intratumoral drugs; quinine dihydrochloride used in single drug group 4 is an intratumoral drug concentration ≥3. Conventional antimalarial drugs used as chemical ablative agents; methylene blue used in single drug group 2 is a conventional vital dye that can only be used as chemical ablative agents when the concentration is greater than or equal to 1. The tumor inhibition rates of single drug groups 2, 3, and 4 were 13%, 53%, and 58%, respectively. However, among the composition groups 5, 6, and 7 corresponding to the single-agent groups 2, 3, and 4, the tumor inhibition rate of the composition group 5 was more than 135% of that of the other two groups.
根据上表的结果，组合物组5、6、7的实际/预期比q分别为>1.15、<1.15和<1.15,显示出明显不同的共用作用。显然，组合物组5、6、7之间的明显药效差异是有无明显协同药效所致。According to the results in the above table, the actual/expected ratios q of composition groups 5, 6, 7 were >1.15, <1.15 and <1.15, respectively, showing clearly different synergistic effects. Apparently, the significant difference in efficacy among the composition groups 5, 6, and 7 was caused by the presence or absence of obvious synergistic efficacy.
根据以上结果及其它类似结果，免疫球蛋白局部活性并未局部协同5-氟尿嘧啶那样的细胞毒抗肿瘤药物、或二盐酸奎宁那样的具有化学消融作用的抗疟药物，却可以局部协同在实验条件下并不显示明显化学消融作用的亚甲蓝。于是，本发明的组合物的局部协同药理并不必须免疫球蛋白具有局部药效，甚至于不必须免疫球蛋白协同物具有有效局部药效。免疫球蛋白协同物与是否显示抗肿瘤细胞有效活性、是否显示化学消融有效活性并无必然关系。该局部协同药理看来与己知的常规病理协同(例如免疫活性协同、抗肿瘤细胞协同、化学消融协同、抗疟协同、等等)均有所不同，而是一种尚待研究的新的协同药理。According to the above and other similar results, the local immunoglobulin activity does not locally synergize with cytotoxic antitumor drugs such as 5-fluorouracil or antimalarial drugs with chemical ablative effects such as quinine dihydrochloride, but can locally synergize in experimental Methylene blue, which does not show significant chemical ablation under conditions. Thus, the local synergistic pharmacology of the composition of the present invention does not necessarily require the immunoglobulin to have local efficacy, or even does not necessarily require the immunoglobulin synergist to have effective local efficacy. There is no necessary relationship between immunoglobulin synergist and whether it shows effective activity against tumor cells or whether it shows effective activity for chemical ablation. This local synergistic pharmacology appears to be different from the known conventional pathological synergy (eg, immune activity synergy, anti-tumor cell synergy, chemical ablation synergy, antimalarial synergy, etc.) Synergistic Pharmacology.
以上研究及更多的类似研究显示，免疫球蛋白提供局部活性或作为局部活性成分(局部协同物)应用的一个必要技术条件是:免疫球蛋白只能与非特定药理的个别药物产生所述局部协同作用。根据以上的结果，所述免疫球蛋白协同物优选为选自抗肿瘤或/和抗全身性病原体(寄生虫、细菌、病毒)的常规无效药物之一种或多种，更优选为选自以下组的一种或多种：氨基酸类营养素、酸消融剂、活体染料。以下实验对免疫球蛋白协同物进行进一步优选。The above studies and more similar studies have shown that a necessary technical condition for immunoglobulins to provide local activity or to be used as a local active ingredient (topical synergist) is that immunoglobulins can only produce said topical with individual drugs of non-specific pharmacology. synergy. According to the above results, the immunoglobulin synergist is preferably one or more conventional ineffective drugs selected from anti-tumor or/and anti-systemic pathogens (parasites, bacteria, viruses), more preferably selected from the following One or more of the group: amino acid nutrient, acid ablating agent, vital dye. The following experiments further optimize the immunoglobulin synergists.
在一个试验中，试验动物为BALB/c小鼠，建模细胞为乳腺癌4T1细胞，以100ul(每毫升1×10 6个细胞)/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(瘤体平均体积319mm 3)随机分为1个阴性对照组(0)和11个药物研究组(1-11)。阴性对照物为生理盐水，研究药物如下表所示。药物均为水溶液，按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。各组均每3日用药一次，一共3次，行瘤内注射，注射量150μl/只/次。用药结束后3日，对动物进行安乐死，解剖后测定瘤重，并从阴性对照组计算抑瘤率，结果示于表8。 In one experiment, the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1×10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model. Successfully modeled experimental animals (average tumor volume 319 mm 3 ) were randomly divided into 1 negative control group (0) and 11 drug study groups (1-11). The negative control is normal saline, and the research drugs are shown in the table below. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each group was administered once every 3 days for a total of 3 times. 3 days after the end of the administration, the animals were euthanized, the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 8.
表8Table 8
Figure PCTCN2022076823-appb-000009
Figure PCTCN2022076823-appb-000009
Figure PCTCN2022076823-appb-000010
Figure PCTCN2022076823-appb-000010
在上表中，组合物组7、8、9、10、11的实际/预期比q均为>1.15，均显示出明显的协同药效。在这些组合物组之间比较，研究组7、9、10、11显示出有效药效(抑瘤率大于40％)，而研究组8未显示出有效药效(抑瘤率小于40％)，且显示有效药效各组分别与研究组8之间的瘤重差异均有统计学意义(均为p<0.05)。In the above table, the actual/expected ratio q of composition groups 7, 8, 9, 10, and 11 are all >1.15, all showing obvious synergistic efficacy. Comparing between these composition groups, study groups 7, 9, 10, 11 showed effective efficacy (tumor inhibition rate greater than 40%), while study group 8 did not show effective efficacy (tumor inhibition rate less than 40%) , and showed that the difference in tumor weight between each group and the study group 8 was statistically significant (all p<0.05).
根据上述研究及更多的类似研究，本发明的免疫球蛋白/氨基酸类营养素组合物中的氨基酸类营养素似乎可选择包含的范围较宽，优选为选自包含以下组中的氨基酸或其盐或者包含或由以下氨基酸构成的寡肽和多肽：精氨酸、甘氨酸、半胱氨酸、丙氨酸、丝氨酸、谷氨酸。According to the above studies and more similar studies, it seems that the amino acid nutrients in the immunoglobulin/amino acid nutrient composition of the present invention can be selected from a wide range, preferably amino acids or their salts selected from the group consisting of: Oligopeptides and polypeptides comprising or consisting of the following amino acids: arginine, glycine, cysteine, alanine, serine, glutamic acid.
在一个试验中，试验动物为BALB/c小鼠，建模细胞为黑色素瘤细胞，以100ul(每毫升2.5×10 6个细胞)/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(瘤体平均体积238mm 3)随机分为1个阴性对照组(0)和9个药物研究组(1-9)。阴性对照物为生理盐水，研究药物如下表所示。药物均为水溶液，按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。各组均每3日用药一次，一共3次，行瘤内注射，注射量125μl/只/次。用药结束后3日，对动物进行安乐死，解剖后测定瘤重，并从阴性对照组计算抑瘤率，结果示于表9。 In one experiment, the experimental animals were BALB/c mice, and the model cells were melanoma cells, and 100ul (2.5×10 6 cells per milliliter) per animal were subcutaneously transplanted into the right armpit of the animal to model the tumor. Successfully modeled experimental animals (average tumor volume 238 mm 3 ) were randomly divided into 1 negative control group (0) and 9 drug study groups (1-9). The negative control is normal saline, and the research drugs are shown in the table below. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each group was administered once every 3 days for a total of 3 times. The animals were euthanized 3 days after the drug administration, and the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 9.
表9Table 9
组别group 药物drug 瘤重(x±s)(g)Tumor weight (x±s) (g) 抑瘤率tumor inhibition rate
00 生理盐水normal saline 2.38±0.262.38±0.26 00
11 4％人免疫球蛋白4% Human Immunoglobulin 2.50±0.242.50±0.24 -5％-5%
22 1％亚甲蓝1% methylene blue 2.02±0.292.02±0.29 15％15%
33 1％新亚甲蓝1% new methylene blue 2.04±0.232.04±0.23 14％14%
44 1％专利蓝1% patent blue 1.93±0.221.93±0.22 19％19%
55 1％孟加拉红1% Red Bengal 2.14±0.192.14±0.19 10％10%
66 4％人免疫球蛋白/1％亚甲蓝4% human immunoglobulin/1% methylene blue 0.69±0.140.69±0.14 71％71%
77 4％人免疫球蛋白/1％新亚甲蓝4% human immunoglobulin/1% new methylene blue 0.62±0.120.62±0.12 74％74%
88 4％人免疫球蛋白/1％专利蓝4% Human Immunoglobulin/1% Patent Blue 1.09±0.171.09±0.17 54％54%
99 4％人免疫球蛋白/1％孟加拉红4% Human Ig/1% Red Bengal 1.95±0.211.95±0.21 18％18%
在上表中，组合物组6、7、8、9的实际/预期比q均为>1.15，均显示出明显的协同药效。在这些组合物组之间比较，研究组6、7、8显示出有效药效(抑瘤率大于40％)，而研究组9未显示出有效药效(抑瘤率小于40％)，且显示有效药效各组分别与研究组9之 间的瘤重差异均有统计学意义(均为p<0.05)。In the above table, the actual/expected ratio q of composition groups 6, 7, 8, and 9 are all >1.15, all showing obvious synergistic efficacy. Comparing between these composition groups, study groups 6, 7, 8 showed effective efficacy (tumor inhibition rate greater than 40%), while study group 9 did not show effective efficacy (tumor inhibition rate less than 40%), and The difference in tumor weight between each group and study group 9 was statistically significant (all p<0.05).
根据上述研究及更多的类似研究，本发明的免疫球蛋白/活体染料组合物中的活体染料优选为选自亚甲蓝类活体染料，更优选为选自亚甲蓝及其衍生物。According to the above studies and more similar studies, the vital dye in the immunoglobulin/vital dye composition of the present invention is preferably selected from methylene blue type vital dyes, more preferably selected from methylene blue and its derivatives.
实施例8：Ig提供局部活性所需条件:Ig药理浓度研究及优选Embodiment 8: Ig provides the conditions required for local activity: Ig pharmacological concentration research and optimization
在一个试验中，试验动物为BALB/c小鼠，建模细胞为乳腺癌4T1细胞，以100ul(每毫升1×10 6个细胞)/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(瘤体平均体积386mm 3)随机分为1个阴性对照组(01)和14个药物研究组(1-14)。阴性对照物为生理盐水，研究药物如下表所示。药物均为水溶液，按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。各实验组均行瘤内注射，每3日用药一次，一共3次，每次注射量如下表10所示。用药结束后3日，对动物进行安乐死，解剖后测定瘤重，并从阴性对照组计算抑瘤率，结果示于表10。 In one experiment, the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1×10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model. The successfully modeled experimental animals (average tumor volume 386mm 3 ) were randomly divided into 1 negative control group (01) and 14 drug study groups (1-14). The negative control is normal saline, and the research drugs are shown in the table below. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each experimental group was injected intratumorally, once every 3 days, for a total of 3 times, and the amount of each injection was shown in Table 10 below. Three days after the end of administration, the animals were euthanized, the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 10.
表10Table 10
Figure PCTCN2022076823-appb-000011
Figure PCTCN2022076823-appb-000011
通常认为，药效取决于剂量而非用药浓度。临床上安全的血药浓度通常非常之低(例如小于10ug/ml)，远低于通常的制剂浓度。常规组合物通常以所含活性成分的量比来表征其组成即可。然而，在上表中，研究组5的抑瘤率是研究组6的1000％以上，研究组9的抑 瘤率是研究组10的200％以上，研究组13的抑瘤率是研究组14的660％以上。It is generally believed that efficacy depends on dose rather than drug concentration. Clinically safe plasma concentrations are usually very low (eg, less than 10 ug/ml), much lower than usual formulation concentrations. Conventional compositions can usually be characterized by the amount ratio of the active ingredients contained. However, in the above table, the tumor inhibition rate of study group 5 was more than 1000% of that of study group 6, the tumor inhibition rate of study group 9 was more than 200% of that of study group 10, and the tumor inhibition rate of study group 13 was more than that of study group 14. 660% or more.
更加重要的是，在局部用药时，相同剂量、相同量比的共用药物之间(例如组合物组5和6之间、9和10之间、13和14之间)可以显示出大不相同的共用效应。组合物组5、9、13的实际/预期比q均为>1.15，都显示出明显的协同药效。而组合物组6、10、14的实际/预期比q均为<1.15，均未显示出协同药效。这些结果说明，本发明的组合物在相同剂量相同量比时可以显示出大不相同的药效，于是有可能显示出大不相同的靶和药理。.More importantly, in topical administration, co-drugs of the same dose and ratio (eg, between composition groups 5 and 6, between 9 and 10, between 13 and 14) can show substantial differences sharing effect. The actual/expected ratio q of composition groups 5, 9 and 13 were all >1.15, all showing obvious synergistic efficacy. However, the actual/expected ratio q of composition groups 6, 10, and 14 were all <1.15, and none of them showed synergistic efficacy. These results indicate that the compositions of the present invention can exhibit greatly different pharmacological effects at the same dose and the same amount ratio, and thus may exhibit greatly different targets and pharmacology. .
实际上，常规用药和局部用药的组成就靶区药学而言具有完全不同的含义。同一药物混合物经常规用药(例如静脉注射)和局部用药(例如瘤內注射)后，将在同一靶区(例如瘤体)形成大不相同的药物混合物。常规用药分布在血液中再随血液到达靶区，在经历复杂的物理、化学、生物过程(例如不同药物组分在血液中的稀释、不同吸附、在某些器官中的不同滞留甚至代谢、等等)后，原混合物以经不复存在。不同的药物组分以不同以往的浓度(往往稀释百倍以上)、甚至不同的复合物而存在于新的血-药混合物中。而药物混合物的局部给药，则是依靠外力直接进入靶区，其在靶区的组成与其给药组成几乎相同。同一药物混合物在靶区中这两种大不相同的组成完全有可能各有其靶从而各有其药理。In fact, the composition of conventional and topical medications has completely different implications in terms of target pharmacy. After the same drug mixture is conventionally administered (eg, intravenous injection) and topical (eg, intratumoral injection), a very different drug mixture will be formed in the same target area (eg, tumor). Conventional drugs are distributed in the blood and then reach the target area with the blood. After undergoing complex physical, chemical and biological processes (such as dilution of different drug components in the blood, different adsorption, different retention in certain organs and even metabolism, etc. etc.), the original mixture no longer exists. Different drug components are present in the new blood-drug mixture in different concentrations (often more than a hundredfold dilution), and even different complexes. The local administration of the drug mixture relies on external force to directly enter the target area, and its composition in the target area is almost the same as its administration composition. It is entirely possible that these two very different compositions of the same drug mixture in the target area have their own targets and thus their own pharmacology.
以上研究及更多的类似研究显示，免疫球蛋白提供局部活性或作为局部活性成分(局部协同物)应用的一个必要技术条件是:所述免疫球蛋白的局部给药浓度大于其产生局部协同的阈值。在本发明申请中，该浓度阈值被称作局部协同浓度。以下试验对该组成条件进行进一步研究。The above studies and more similar studies have shown that a necessary technical condition for immunoglobulin to provide local activity or to be used as a local active ingredient (topical synergist) is that the immunoglobulin is administered locally at a concentration greater than it produces local synergy. threshold. In the present application, this concentration threshold is referred to as the local synergistic concentration. The following tests further investigate this compositional condition.
在一个试验中，试验动物为BALB/c小鼠，建模细胞为乳腺癌4T1细胞，以100ul(每毫升1×10 6个细胞)/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(瘤体平均体积297mm 3)随机分为1个阴性对照组(0)和7个药物研究组(1-7)。阴性对照物为生理盐水，研究药物如下表所示。药物均为水溶液，按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。各组均每3日用药一次，一共3次，行瘤内注射，每次注射量如下表11所示。用药结束后3日，对动物进行安乐死，解剖后测定瘤重，并从阴性对照组计算抑瘤率，结果示于表11。 In one experiment, the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1×10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model. Successfully modeled experimental animals (average tumor volume 297mm 3 ) were randomly divided into 1 negative control group (0) and 7 drug study groups (1-7). The negative control is normal saline, and the research drugs are shown in the table below. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each group was administered once every 3 days, a total of 3 times, and intratumoral injection was performed, and the amount of each injection was shown in Table 11 below. Three days after the end of administration, the animals were euthanized, the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 11.
表11Table 11
Figure PCTCN2022076823-appb-000012
Figure PCTCN2022076823-appb-000012
在上表中，在免疫球蛋白给药浓度为<2％的区间内，组合物组3、4、5与阴性对照组01之间的瘤重差异均无统计学意义(均为p>0.05)，免疫球蛋白未显示出与精氨酸的局部协同作用。In the above table, in the interval of immunoglobulin administration concentration <2%, there was no significant difference in tumor weight between the composition group 3, 4, 5 and the negative control group 01 (all p>0.05 ), immunoglobulins did not show local synergy with arginine.
然而，当组合物中免疫球蛋白给药浓度达到一个阈值(≥2％)时，组合物组6与阴性对照组01之间的瘤重差异有统计学意义(p<0.05)、且其实际/预期比q为>1.15，显示出明显协同药效。组合物组7进一步证实了该阈值的出现并非偶然。在相同的免疫球蛋白剂量下，其抑瘤率随免疫球蛋白浓度提高进一步上升，显示出更高的协同药效。However, when the administration concentration of immunoglobulin in the composition reached a threshold (≥2%), the difference in tumor weight between composition group 6 and negative control group 01 was statistically significant (p<0.05), and its actual The /expected ratio q is >1.15, showing a clear synergistic effect. Composition group 7 further confirms that the occurrence of this threshold is not accidental. At the same dose of immunoglobulin, the tumor inhibition rate further increased with the increase of the concentration of immunoglobulin, showing a higher synergistic effect.
在一个试验中，试验动物为BALB/c小鼠，建模细胞为乳腺癌4T1细胞，以0.5×10 5个细胞/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(瘤体平均体积259±32mm 3)随机分为1个阴性对照组(0)和7个药物研究组(1-7)。阴性对照物为生理盐水，研究药物如下表所示。药物均为水溶液，按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。各组均每3日用药一次，一共3次，行瘤内注射，每次注射量如下表所示。第一次用药第7日和第21日，分别测定各系列各组别动物的瘤体体积，并从各系列阴性对照组分别计算1-4组的7日体积抑瘤率(r 7)和21日体积抑瘤率(r 21)，结果示于下表12。 In one experiment, the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 0.5×10 5 cells per animal were subcutaneously transplanted into the right armpit of the animal to model. Successfully modeled experimental animals (average tumor volume 259±32 mm 3 ) were randomly divided into 1 negative control group (0) and 7 drug study groups (1-7). The negative control is normal saline, and the research drugs are shown in the table below. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each group was administered once every 3 days, a total of 3 times, and intratumoral injection was performed, and the amount of each injection was shown in the table below. On the 7th and 21st days of the first administration, the tumor volume of animals in each series and each group was measured, and the 7-day volume tumor inhibition rate (r 7 ) and The volume tumor inhibition rate (r 21 ) on the 21st day is shown in Table 12 below.
表12不同药物组别对应的体积抑瘤率(r 7、r 21)结果 Table 12 Results of volume tumor inhibition rate (r 7 , r 21 ) corresponding to different drug groups
Figure PCTCN2022076823-appb-000013
Figure PCTCN2022076823-appb-000013
在上表中,通过组别1、2、5,1、3、6,和1、4、7的r 7和r 21计算出的人免疫球蛋白/亚甲蓝组合物的q 7和q 21均为>1.15,说明高浓度Ig的局部活性提供了如以上实施例2-4中所述的超过局部加和作用预期的短期协同作用和中长期协同作用。此外,q 21随Ig浓度升高而有所上升,说明该中长期协同作用有一定的Ig浓度依赖性。 In the table above, q 7 and q of the human immunoglobulin/methylene blue composition calculated by r 7 and r 21 of groups 1, 2, 5, 1, 3, 6, and 1, 4, 7 21 are all >1.15, indicating that the local activity of high concentrations of Ig provides both short-term synergy and mid- to long-term synergy beyond the expectations of local additive effects as described in Examples 2-4 above. In addition, q 21 increased with the increase of Ig concentration, which indicated that the mid- and long-term synergy had a certain Ig concentration dependence.
根据上述研究及更多的类似研究，免疫球蛋白提供局部活性或作为本发明的组合物中的局部协同物的一个必要条件为：所述免疫球蛋白的浓度必须为>1.5％、优选为2.0％-30％或3.0％-30％。所述免疫球蛋白在局部给药浓度小于该协同浓度时可能作为常规的免疫药物、而不能作为本发明中的局部活性成分应用于制备用于治疗局部病变疾病的局部药物。According to the above studies and more similar studies, a necessary condition for an immunoglobulin to provide local activity or to act as a local synergist in the compositions of the present invention is that the concentration of the immunoglobulin must be >1.5%, preferably 2.0 %-30% or 3.0%-30%. When the local administration concentration is less than the synergistic concentration, the immunoglobulin may be used as a conventional immune drug, but cannot be used as a topical active ingredient in the present invention to prepare a topical drug for the treatment of local pathological diseases.
实施例9：Ig提供局部活性所需条件:共用物药理浓度研究及优选Embodiment 9: Ig provides the conditions required for local activity: the study and optimization of the pharmacological concentration of the concomitant
在一个试验中，试验动物为BALB/c小鼠，建模细胞为乳腺癌4T1细胞，以100ul(每毫升1×10 6个细胞)/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(瘤体平均体积311mm 3)随机分为1个阴性对照组(0)和9个药物研究组(1-9)。阴性对照物为生理盐水，研究药物如下表所示。药物均为水溶液，按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。各组均每3日用药一次，一共3次，行瘤内注射，每次注射量如下表所示。用药结束后3日，对动物进行安乐死，解剖后测定瘤重，并从阴性对照组计算抑瘤率，结果示于表13。 In one experiment, the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1×10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model. Successfully modeled experimental animals (average tumor volume 311 mm 3 ) were randomly divided into 1 negative control group (0) and 9 drug study groups (1-9). The negative control is normal saline, and the research drugs are shown in the table below. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each group was administered once every 3 days, a total of 3 times, and intratumoral injection was performed, and the amount of each injection was shown in the table below. Three days after the end of administration, the animals were euthanized, the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 13.
表13Table 13
Figure PCTCN2022076823-appb-000014
Figure PCTCN2022076823-appb-000014
Figure PCTCN2022076823-appb-000015
Figure PCTCN2022076823-appb-000015
在上表中，在还原型谷胱甘肽给药浓度为<8％的区间内，组合物组6和7与阴性对照组01之间的瘤重差异均无统计学意义(均为p>0.05)，免疫球蛋白未显示出与还原型谷胱甘肽的局部协同作用。In the above table, in the interval where the administration concentration of reduced glutathione was <8%, there was no significant difference in tumor weight between the composition groups 6 and 7 and the negative control group 01 (both p> 0.05), the immunoglobulin did not show local synergy with reduced glutathione.
然而，当组合物中还原型谷胱甘肽给药浓度达到一个阈值(≥8％)时，组合物组8与阴性对照组01之间的瘤重差异有统计学意义(p<0.05)、且其实际/预期比q为>1.15，显示出明显协同药效。组合物组9进一步证实了该阈值的出现并非偶然。在相同的还原型谷胱甘肽剂量下，其抑瘤率随还原型谷胱甘肽浓度提高进一步上升，显示出更高的协同药效。此外,在其它实施例中还显示所述免疫球蛋白与超过上述浓度阈值(例如15％、20％等)的其它氨基酸(例如精氨酸、甘氨酸等)的组合物的局部协同作用。However, when the administration concentration of reduced glutathione in the composition reached a threshold (≥8%), the difference in tumor weight between composition group 8 and negative control group 01 was statistically significant (p<0.05), And its actual/expected ratio q is >1.15, showing obvious synergistic effect. Composition group 9 further confirms that the occurrence of this threshold is not accidental. At the same dose of reduced glutathione, the tumor inhibition rate further increased with the increase of the concentration of reduced glutathione, showing a higher synergistic effect. In addition, local synergistic effects of the immunoglobulin compositions with other amino acids (eg, arginine, glycine, etc.) above the aforementioned concentration thresholds (eg, 15%, 20%, etc.) are also shown in other examples.
根据上述研究及更多的类似研究，免疫球蛋白提供局部活性使得氨基酸类营养素作为其局部协同物的一个必要条件为免疫球蛋白和氨基酸类营养素的浓度比而非量比满足下述条件：所述免疫球蛋白的浓度必须如以上实施例8中的优选浓度,而所述氨基酸类营养素的浓度必须为>7％、≥8％、优选为8-30％或10-30％。所述氨基酸类营养素在局部给药浓度小于该协同浓度时可能作为免疫球蛋白的助剂(增溶剂、稳定剂、等等)、而不能作为本发明中的免疫球蛋白的局部协同物应用于制备用于治疗局部病变疾病的局部药物。According to the above studies and many more similar studies, a necessary condition for immunoglobulin to provide local activity such that amino acid nutrients act as their local synergists is that the concentration ratio, not the quantitative ratio, of immunoglobulin and amino acid nutrients satisfies the following conditions: The concentration of the immunoglobulin must be the preferred concentration in Example 8 above, and the concentration of the amino acid nutrients must be >7%, ≥8%, preferably 8-30% or 10-30%. The amino acid nutrients may be used as adjuvants (solubilizers, stabilizers, etc.) of immunoglobulin when the local administration concentration is less than the synergistic concentration, but cannot be used as the local synergistic agent of immunoglobulin in the present invention. Preparation of topical medicaments for the treatment of local pathological diseases.
在一个试验中，试验动物为BALB/c小鼠，建模细胞为乳腺癌4T1细胞，以100ul(每毫升1×10 6个细胞)/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(瘤体平均体积307mm 3)随机分为1个阴性对照组(0)和9个药物研究组(1-9)。阴性对照物为生理盐水，研究药物如下表所示。药物均为水溶液，按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。各组均每3日用药一次，一共3次，行瘤内注射，每次注射量如下表所示。用药结束后3日，对动物进行安乐死，解剖后测定瘤重，并从阴性对照组计算抑瘤率，结果示于表14。 In one experiment, the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1×10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model. Successfully modeled experimental animals (with an average tumor volume of 307 mm 3 ) were randomly divided into 1 negative control group (0) and 9 drug study groups (1-9). The negative control is normal saline, and the research drugs are shown in the table below. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each group was administered once every 3 days, a total of 3 times, and intratumoral injection was performed, and the amount of each injection was shown in the table below. Three days after the end of administration, the animals were euthanized, the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 14.
表14Table 14
Figure PCTCN2022076823-appb-000016
Figure PCTCN2022076823-appb-000016
在上表中，在亚甲蓝给药浓度为<0.35％的区间内，组合物组6和7与阴性对照组01之间的瘤重差异均无统计学意义(均为p>0.05)，免疫球蛋白未显示出与亚甲蓝的局部协同作用。In the above table, in the interval of methylene blue administration concentration <0.35%, there was no significant difference in tumor weight between composition groups 6 and 7 and negative control group 01 (both p>0.05), Immunoglobulin did not show local synergy with methylene blue.
然而，当组合物中亚甲蓝给药浓度达到一个阈值(≥0.35％)时，组合物组8与阴性对照组01之间的瘤重差异有统计学意义(p<0.05)、且其实际/预期比q为>1.15，显示出明显协同药效。组合物组9进一步证实了该阈值的出现并非偶然。在相同的亚甲蓝剂量下，其抑瘤率随亚甲蓝浓度提高进一步上升，显示出更高的协同药效。However, when the administration concentration of methylene blue in the composition reached a threshold (≥0.35%), the difference in tumor weight between composition group 8 and negative control group 01 was statistically significant (p<0.05), and its actual The /expected ratio q is >1.15, showing a clear synergistic effect. Composition group 9 further confirms that the occurrence of this threshold is not accidental. At the same methylene blue dose, the tumor inhibition rate further increased with the increase of methylene blue concentration, showing a higher synergistic effect.
在一个试验中，试验动物为裸小鼠，建模细胞为人淋巴Raji细胞，以1×10 6个细胞/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(瘤体平均体积127±19mm 3)随机分为1个阴性对照组(0)和5个药物研究组(1-5)。阴性对照物为生理盐水，研究药物如下表所示。药物均为水溶液，按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。各组均每3日用药一次，一共3次，行瘤内注射，每次注射量如下表所示。第一次用药第7日和第21日，分别测定各系列各组别动物的瘤体体积，并从各系列阴性对照组分别计算1-4组的7日体积抑瘤率(r 7)和21日体积抑瘤率(r 21)，结果示于下表15。 In one experiment, the experimental animals were nude mice, the model cells were human lymphoid Raji cells, and 1×10 6 cells per animal were used to model the transplanted tumor subcutaneously in the right armpit of the animal. Successfully modeled experimental animals (average tumor volume 127±19 mm 3 ) were randomly divided into 1 negative control group (0) and 5 drug study groups (1-5). The negative control is normal saline, and the research drugs are shown in the table below. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each group was administered once every 3 days, a total of 3 times, and intratumoral injection was performed, and the amount of each injection was shown in the table below. On the 7th and 21st days of the first administration, the tumor volume of animals in each series and each group was measured, and the 7-day volume tumor inhibition rate (r 7 ) and The volume tumor inhibition rate (r 21 ) on the 21st day is shown in Table 15 below.
表15不同药物组别对应的体积抑瘤率(r 7、r 21)结果 Table 15 Results of volume tumor inhibition rate (r 7 , r 21 ) corresponding to different drug groups
组别group 药物drug r 7 r 7 r 21 r 21
11 10％人免疫球蛋白10% Human Immunoglobulin -1.3％-1.3% 2.4％2.4%
22 3％亚甲蓝3% methylene blue 61.3％61.3% 22.7％22.7%
33 5％亚甲蓝5% methylene blue 71.5％71.5% 24.1％24.1%
44 10％人免疫球蛋白/3％亚甲蓝10% human immunoglobulin/3% methylene blue 72.5％72.5% 51.5％51.5%
55 10％人免疫球蛋白/5％亚甲蓝10% human immunoglobulin/5% methylene blue 78.3％78.3% 67.3％67.3%
在上表中,通过组别1、2、4,和1、3、5的r 7和r 21计算出的人免疫球蛋白/亚甲蓝组合物的q 7和q 21均为>1.15,说明高浓度Ig的局部活性提供了如以上实施例2-4中所述的超过局部加和作用预期的短期协同作用和中长期协同作用。此外，q 21随Ig浓度升高而有所上升，说明该中长期协同作用有一定的Ig浓度依赖性。 In the above table, the q7 and q21 of the human immunoglobulin/methylene blue composition calculated by the r7 and r21 of groups 1 , 2 , 4, and 1, 3, 5 are both >1.15, It is demonstrated that the local activity of high concentrations of Ig provides both short-term synergy and mid- to long-term synergy that exceeds the expectations of local additive effects as described in Examples 2-4 above. In addition, q 21 increased with the increase of Ig concentration, indicating that the mid- and long-term synergy had a certain Ig concentration dependence.
根据上述研究及更多的类似研究，免疫球蛋白提供局部活性使得活体染料作为其局部协同物的一个必要条件为免疫球蛋白和活体染料的浓度比而非量比满足下述条件：所述免疫球蛋白的浓度必须如以上实施例8中的优选浓度,而所述活体染料的浓度必须为>0.3％、≥0.35％、优选为0.35-10％。所述活体染料在局部给药浓度小于该协同浓度时可能作为免疫球蛋白的助剂、而不能作为本发明中的免疫球蛋白的局部协同物应用于制备用于治疗局部病变疾病的局部药物。According to the above studies and many more similar studies, a necessary condition for immunoglobulin to provide local activity such that vital dye acts as its local synergist is that the concentration ratio of immunoglobulin and vital dye, rather than the amount ratio, satisfies the following condition: The concentration of globulin must be the preferred concentration in Example 8 above, while the concentration of the vital dye must be >0.3%, ≥0.35%, preferably 0.35-10%. When the local administration concentration is less than the synergistic concentration, the vital dye may be used as an adjuvant of immunoglobulin, but cannot be used as a local synergist of immunoglobulin in the present invention to prepare a topical drug for the treatment of local pathological diseases.
实施例10：Ig提供局部活性所需条件:多组分局部协同物研究及优选Example 10: Conditions required for Ig to provide topical activity: research and optimization of multi-component topical synergists
百年来科学界进行了数不清的组合物研究，而临床上可用的混合型组合物则是少之又少。通常认为，以单药与单药混合在一起去获得抗肿瘤协同药效是一个小概率事件。进一步，显示出该协同药效的组合物与单药进行二次组合以获得新的抗肿瘤协同药效则更是难之又难，犹其是这些单药还局限于选自先前的协同物中。Countless composition studies have been carried out by the scientific community for hundreds of years, and clinically available hybrid compositions are few and far between. It is generally believed that it is a small probability event that a single drug is mixed with a single drug to obtain an anti-tumor synergistic effect. Further, it is even more difficult to obtain a new anti-tumor synergistic effect by combining the composition showing this synergistic effect twice with a single drug, even though these single drugs are still limited to selected from the previous synergists. middle.
在一个试验中，试验动物为BALB/c小鼠，建模细胞为乳腺癌4T1细胞，以100ul(每毫升1×10 6个细胞)/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(瘤体平均体积320mm 3)随机分为1个阴性对照组(0)和9个药物研究组(1-9)。阴性对照物为生理盐水，研究药物如下表所示。药物均为水溶液，按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。各组均每3日用药一次，一共3次，行瘤内注射，每次注射量如下表所示。用药结束后3日，对动物进行安乐死，解剖后测定瘤重，并从阴性对照组计算抑瘤率，结果示于表16。 In one experiment, the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1×10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model. Successfully modeled experimental animals (average tumor volume 320mm 3 ) were randomly divided into 1 negative control group (0) and 9 drug study groups (1-9). The negative control is normal saline, and the research drugs are shown in the table below. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each group was administered once every 3 days, a total of 3 times, and intratumoral injection was performed, and the amount of each injection was shown in the table below. Three days after the end of administration, the animals were euthanized, the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 16.
表16Table 16
Figure PCTCN2022076823-appb-000017
Figure PCTCN2022076823-appb-000017
Figure PCTCN2022076823-appb-000018
Figure PCTCN2022076823-appb-000018
在上表中，在乙酸给药浓度为<2％的区间内，组合物组6和7与阴性对照组01之间的瘤重差异均无统计学意义(均为p>0.05)，免疫球蛋白未显示出与甘氨酸/乙酸的局部协同作用。In the above table, in the interval of acetic acid administration concentration of <2%, there was no significant difference in tumor weight between the composition groups 6 and 7 and the negative control group 01 (both p>0.05). The protein did not show local synergy with glycine/acetate.
然而，当组合物中乙酸给药浓度达到一个阈值(≥2％)时，组合物组8与阴性对照组01之间的瘤重差异有统计学意义(p<0.05)、且其实际/预期比q为>1.15，显示出明显协同药效。组合物组9进一步证实了该阈值的出现并非偶然。在相同的甘氨酸/乙酸剂量下，其抑瘤率随乙酸浓度提高进一步上升，显示出更高的协同药效。However, when the administration concentration of acetic acid in the composition reached a threshold (≥2%), the difference in tumor weight between composition group 8 and negative control group 01 was statistically significant (p<0.05), and its actual/expected The ratio q is >1.15, showing obvious synergistic effect. Composition group 9 further confirms that the occurrence of this threshold is not accidental. At the same dose of glycine/acetic acid, the tumor inhibition rate further increased with the increase of the concentration of acetic acid, showing a higher synergistic effect.
根据上述研究及更多的类似研究，氨基酸类营养素/酸消融剂作为本发明的组合物中的免疫球蛋白的局部协同物的一个必要条件为：所述氨基酸类营养素的浓度为其在实施例7中的所述局部协同浓度，所述酸消融剂必须为大于其单药即可显示出化学消融药效的浓度，例如强酸为≥0.5％、弱酸或弱碱(含氮弱碱、弱酸强碱盐)为≥2.0％或≥2.5％。所述酸消融剂为弱酸时，其在本发明的组合物中的浓度为为≥2.0％、≥3.0％、优选为3.0-15％或5-15％。According to the above studies and more similar studies, a necessary condition for the amino acid nutrient/acid ablative agent as a topical synergist for immunoglobulins in the compositions of the present invention is that the amino acid nutrient is at a concentration as described in the Examples For the local synergistic concentration in 7, the acid ablating agent must be a concentration greater than that of its single drug to show chemical ablation efficacy, such as strong acid ≥ 0.5%, weak acid or weak base (nitrogen-containing weak base, weak acid strong base salt) is ≥ 2.0% or ≥ 2.5%. When the acid ablating agent is a weak acid, its concentration in the composition of the present invention is ≥2.0%, ≥3.0%, preferably 3.0-15% or 5-15%.
更多的类似研究结果说明，而本发明的组合物的协同作用可以在其中的酸消融剂浓度在远小于(例如<50％、<30％、优选为20％-30％)其单药临床常用浓度(例如乙酸为50％、氢氧化钠为7％、盐酸为5％)条件下获得相同、甚至更高的药效。因而，本发明的组合物的协同作用既包括药效协同作用又包括安全性协同作用。The results of more similar studies show that the synergistic effect of the composition of the present invention can be achieved in that the concentration of the acid ablating agent is much lower than (eg <50%, <30%, preferably 20%-30%) of its single-agent clinical The same or even higher efficacy is obtained at common concentrations (eg, 50% acetic acid, 7% sodium hydroxide, and 5% hydrochloric acid). Thus, the synergistic effect of the composition of the present invention includes both pharmacodynamic synergy and safety synergy.
在一个试验中，试验动物为BALB/c小鼠，建模细胞为乳腺癌4T1细胞，以100ul(每毫升1×10 6个细胞)/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(瘤体平均体积307mm 3)随机分为1个阴性对照组(0)和10个药物研究组(1-10)。阴性对照物为生理盐水，研究药物如下表所示。药物均为水溶液，按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。各组均每3日用药一次，一共3次，行瘤 内注射，每次注射量100μl/只。用药结束后3日，对动物进行安乐死，解剖后测定瘤重，并从阴性对照组计算抑瘤率，结果示于表17。 In one experiment, the experimental animals were BALB/c mice, and the model cells were breast cancer 4T1 cells, and 100ul (1×10 6 cells per milliliter) per animal were transplanted subcutaneously in the right armpit of the animal to model. Successfully modeled experimental animals (average tumor volume 307 mm 3 ) were randomly divided into 1 negative control group (0) and 10 drug study groups (1-10). The negative control is normal saline, and the research drugs are shown in the table below. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each group was administered once every 3 days for a total of 3 times. Three days after the end of the administration, the animals were euthanized, the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group. The results are shown in Table 17.
表17Table 17
Figure PCTCN2022076823-appb-000019
Figure PCTCN2022076823-appb-000019
在上表中，组合物组4和5与阴性对照组01之间的瘤重差异均有统计学意义(均为p<0.05)、且其实际/预期比q均为>1.15，均显示出明显协同药效。In the above table, the difference in tumor weight between the composition groups 4 and 5 and the negative control group 01 was statistically significant (both p<0.05), and the actual/expected ratio q was >1.15, all showing that Significant synergistic effect.
在一个试验中，试验动物为BALB/c小鼠，建模细胞为乳腺癌4T1细胞，以100ul(每毫升0.5×10 6个细胞)/只在动物右侧腋部皮下进行移植瘤建模。成功建模的试验动物(瘤体平均体积237±26mm 3)随机分为1个阴性对照组(0)和7个药物研究组(1-7)。阴性对照物为生理盐水，研究药物如下表所示,其中的Ig共用物为亚甲蓝与聚多糖的组合物,所述聚多糖的模型药物分别为羧甲基葡聚糖(酵母菌源)、水溶性β-D-葡聚糖(酵母菌源)、和右旋糖酐。药物均为水溶液，按实施例1的制备方法配置而成,其中的人免疫球蛋白为表1中的冻干静注人免疫球蛋白。各组均每2日用药一次，一共2次，行瘤内注射，每次注射量为100ul。第一次用药第7日和第21日，分别测定各系列各组别动物的瘤体体积，并相对于各系列阴性对照组分别计算1-4组的7日体积抑瘤率(r 7)和34日体积抑瘤率(r 21)，结果示于下表18。 In one experiment, the experimental animals were BALB/c mice, the model cells were breast cancer 4T1 cells, and 100ul (0.5×10 6 cells per milliliter) per animal was used to model the transplanted tumor subcutaneously in the right armpit of the animal. Successfully modeled experimental animals (average tumor volume 237±26 mm 3 ) were randomly divided into 1 negative control group (0) and 7 drug study groups (1-7). The negative control is normal saline, the research drugs are shown in the following table, the Ig compound is the composition of methylene blue and polysaccharide, and the model drug of the polysaccharide is carboxymethyl glucan (yeast source) , water-soluble beta-D-glucan (yeast source), and dextran. The medicines are all aqueous solutions, prepared according to the preparation method of Example 1, and the human immunoglobulins in them are the freeze-dried intravenous human immunoglobulins in Table 1. Each group was administered once every 2 days, a total of 2 times, intratumoral injection, each injection volume was 100ul. On the 7th and 21st days of the first administration, the tumor volume of animals in each series and each group was measured, and the 7-day volume tumor inhibition rate (r 7 ) of groups 1-4 was calculated relative to the negative control group of each series. and 34-day volume tumor inhibition rate (r 21 ), the results are shown in Table 18 below.
表18Table 18
组别group 药物drug r 7 r 7 r 34 r 34
11 8％人免疫球蛋白8% Human Immunoglobulin 0.4％0.4% -0.9％-0.9%
22 2％亚甲蓝/10％羧甲基葡聚糖2% methylene blue/10% carboxymethyl dextran 79.8％79.8% 32.1％32.1%
33 2％亚甲蓝/15％水溶性β-D-葡聚糖2% methylene blue/15% water soluble beta-D-glucan 72.6％72.6% 28.9％28.9%
44 2％亚甲蓝/1.5％右旋糖酐702% methylene blue/1.5% dextran 70 58.6％58.6% 31.5％31.5%
55 8％人免疫球蛋白/2％亚甲蓝/10％羧甲基葡聚糖8% human immunoglobulin/2% methylene blue/10% carboxymethyl dextran 65.7％65.7% 48.6％48.6%
66 8％人免疫球蛋白/2％亚甲蓝/15％水溶性β-D-葡聚糖8% Human Immunoglobulin/2% Methylene Blue/15% Water Soluble Beta-D-Glucan 61.5％61.5% 43.8％43.8%
77 8％人免疫球蛋白/2％亚甲蓝/1.5％右旋糖酐708% Human Immunoglobulin/2% Methylene Blue/1.5% Dextran 70 60.4％60.4% 38.2％38.2%
在上表中,通过组1、2、5的r 7和r 21计算出的组别5组合物的q 7和q 21分别为<1.00和>1.15；通过组1、3、6的r 7和r 21计算出的组别6组合物的q 7和q 21分别为<1.00和>1.15；通过组1、 4、7的r 7和r 21计算出的组别7组合物的q 7和q 21分别为<1.00和>1.15,说明Ig局部活性在未能提供短期协同作用的条件下提供了超过中长期加和作用预期的中长期协同作用。 In the table above, the q7 and q21 of the Group 5 composition calculated by the r7 and r21 of the Groups 1 , 2, 5 are <1.00 and >1.15, respectively; by the r7 of the Groups 1, 3, 6 Calculated q7 and q21 for Group 6 compositions were <1.00 and >1.15, respectively; q7 and q21 for Group 7 compositions calculated from r7 and r21 for Groups 1, 4 , 7 q 21 were <1.00 and >1.15, respectively, indicating that Ig local activity provided mid- and long-term synergy beyond that expected for mid- and long-term additive effects under conditions that failed to provide short-term synergy.
以上研究及更多的类似研究显示，免疫球蛋白作为局部协同物应用的一个优选技术条件是:所述组合物包含免疫球蛋白和2种或2种以上的多种免疫球蛋白协同物。The above studies and more similar studies have shown that a preferred technical condition for the application of immunoglobulin as a topical synergist is: the composition comprises immunoglobulin and two or more multiple immunoglobulin synergists.
除本文中描述的那些外，根据前述描述，本发明的多种修改对本领域技术人员而言会是显而易见的。这样的修改也意图落入所附权利要求书的范围内。本申请中所引用的各参考文献(包括所有专利、专利申请、期刊文章、书籍及任何其它公开)均以其整体援引加入本文。Various modifications of the invention, in addition to those described herein, will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference cited in this application, including all patents, patent applications, journal articles, books, and any other publications, is incorporated by reference in its entirety.

Claims (26)

  1. 免疫球蛋白作为局部活性成分在制备可致靶区组织坏死的药物组合物中的应用,其中所述药物组合物还包含所述免疫球蛋白的局部协同物、以及药物学可接受的合适载体。The application of immunoglobulin as a topical active ingredient in the preparation of a pharmaceutical composition that can cause tissue necrosis in a target area, wherein the pharmaceutical composition further comprises a topical synergist of the immunoglobulin and a suitable pharmaceutically acceptable carrier.
  2. 可致靶区组织坏死的药物组合物，其包含可提供局部活性的免疫球蛋白、所述免疫球蛋白的局部协同物、以及药物学可接受的合适载体。A pharmaceutical composition capable of causing tissue necrosis in a target area, comprising an immunoglobulin that provides local activity, a local synergist of the immunoglobulin, and a suitable pharmaceutically acceptable carrier.
  3. 抗局部病变疾病方法，其包括向受试者局部病变施用可致靶区组织坏死的药物组合物,其中所述药物组合物包含可提供局部活性的免疫球蛋白、所述免疫球蛋白的局部协同物、以及药物学可接受的合适载体。Anti-local pathological disease method, which comprises administering to the local lesions of a subject a pharmaceutical composition that can cause tissue necrosis in a target area, wherein the pharmaceutical composition comprises an immunoglobulin that can provide local activity, a local synergy of the immunoglobulin substance, and a suitable pharmaceutically acceptable carrier.
  4. 根据权利要求1-3之一的应用、药物组合物或方法，其中所述药物组合物为液体剂型,优选地，所述载体包括水。The use, pharmaceutical composition or method according to any one of claims 1-3, wherein the pharmaceutical composition is in a liquid dosage form, preferably the carrier comprises water.
  5. 根据权利要求1-4之一的应用、药物组合物或方法，其中所述药物组合物如此制备、组成或施用以满足所述免疫球蛋白提供所述局部活性所需的以下条件：所述免疫球蛋白以浓度(w/v)为≥2.0，优选地2.0-35％，更优选地3.0-30％进入所述靶区。The use, pharmaceutical composition or method according to any one of claims 1 to 4, wherein said pharmaceutical composition is prepared, composed or administered so as to satisfy the following conditions required for said immunoglobulin to provide said local activity: said immune The globulin enters the target zone at a concentration (w/v) of > 2.0, preferably 2.0-35%, more preferably 3.0-30%.
  6. 根据权利要求5的应用、药物组合物或方法，其中所述免疫球蛋白为人IgG或其工程类似物。The use, pharmaceutical composition or method according to claim 5, wherein the immunoglobulin is human IgG or an engineered analog thereof.
  7. 根据权利要求5或6的应用、药物组合物或方法，其中所述免疫球蛋白提供所述局部活性所需的条件还包括：免疫球蛋白的局部协同物选自针对所述靶区的常规无效药物之一种或多种,且所述免疫球蛋白与所述局部协同物以以下重量比(W 局部协同物/W 免疫球蛋白)进入所述靶区：W 局部协同物/W 免疫球蛋白为≥(0.5-39)/2.0、(0.5-39)/(2.0-35)、或(1-39)/(2.0-35)。 The use, pharmaceutical composition or method according to claim 5 or 6, wherein the conditions required for the immunoglobulin to provide the local activity further comprise: the local synergist of the immunoglobulin is selected from conventional ineffective against the target region One or more of the drugs, and the immunoglobulin and the topical synergist enter the target area in the following weight ratio (W topical synergist /W immunoglobulin ): W topical synergist /W immunoglobulin is >(0.5-39)/2.0, (0.5-39)/(2.0-35), or (1-39)/(2.0-35).
  8. 根据权利要求5或7的应用、药物组合物或方法，其中所述常规无效药物为选自包括以下组的一种或多种：氨基酸类营养素、酸消融剂、活体染料、聚多糖,且所述免疫球蛋白提供所述局部活性所需条件还包括：所述局部协同物以以下给药浓度(W 局部协同物/V 组合物)进入所述靶区：W 局部协同物/V 组合物为>0.35％-39％、≥0.5％-39％、0.5％-39％、或1％-39％。 The application, pharmaceutical composition or method according to claim 5 or 7, wherein the conventional ineffective drug is one or more selected from the group consisting of amino acid nutrients, acid ablating agents, vital dyes, polysaccharides, and all The conditions required for the immunoglobulin to provide the topical activity also include: the topical synergist enters the target area at the following administration concentration (with the topical synergist /V composition ): with the topical synergist /V composition as >0.35%-39%, >0.5%-39%, 0.5%-39%, or 1%-39%.
  9. 根据权利要求8的应用、药物组合物或方法，其中所述常规无效药物为选自以下之组的一种或多种：氨基酸类营养素/酸消融剂、氨基酸类营养素/活体染料、活体染料/酸消融剂、活体染料/氨基酸类营养素/酸消融剂、活体染料/聚多糖。The use, pharmaceutical composition or method according to claim 8, wherein the conventional ineffective drug is one or more selected from the group consisting of amino acid nutrients/acid ablating agents, amino acid nutrients/vital dyes, vital dyes/ Acid ablatives, vital dyes/amino acid nutrients/acid ablatives, vital dyes/polysaccharides.
  10. 根据权利要求8或9的应用、药物组合物或方法，其中所述氨基酸类营养素的施用浓度(w/v)为≥8％、优选为8-30％、更优选为10-20％。The use, pharmaceutical composition or method according to claim 8 or 9, wherein the amino acid nutrient is administered at a concentration (w/v) of ≥8%, preferably 8-30%, more preferably 10-20%.
  11. 根据权利要求8或9的应用、药物组合物或方法，其中所述活体染料的施用浓度(w/v)为≥0.35％、优选为0.35-10％、更优选为1％-5％。The use, pharmaceutical composition or method according to claim 8 or 9, wherein the vital dye is administered at a concentration (w/v) of > 0.35%, preferably 0.35-10%, more preferably 1%-5%.
  12. 根据权利要求8或9的应用、药物组合物或方法，其中所述酸消融剂的施用浓度(w/v)为≥0.5％、优选为0.5-10％或3.5-10％。The use, pharmaceutical composition or method according to claim 8 or 9, wherein the acid ablating agent is administered at a concentration (w/v) of > 0.5%, preferably 0.5-10% or 3.5-10%.
  13. 根据权利要求8-10之一的应用、药物组合物或方法，其中所述氨基酸类营养素包 括或选自具有营养保健效应的以下氨基酸类化合物之一种或多种：氨基酸、氨基酸盐、寡肽和多肽；优选为选自以下组中的氨基酸或其盐或者包含或由以下氨基酸构成的寡肽和多肽：丙氨酸、缬氨酸、亮氨酸、异亮氨酸、苯丙氨酸、脯氨酸、色氨酸、酪氨酸、丝氨酸、半胱氨酸、蛋氨酸、天冬酰胺、谷氨酰胺、苏氨酸、赖氨酸、精氨酸、组氨酸、天冬氨酸、谷氨酸、β-丙氨酸、牛磺酸、γ氨基丁酸(GABA)、茶多酚(茶氨酸)、南瓜子氨基酸(3-氨基-3-羧基吡烷酸)、谷氨酰胺、瓜氨酸、鸟氨酸；更优选为选自以下组中的氨基酸或其盐或者包含或由以下氨基酸构成的寡肽和多肽：精氨酸、赖氨酸、甘氨酸、半胱氨酸、丙氨酸、丝氨酸、谷氨酸。The application, pharmaceutical composition or method according to any one of claims 8 to 10, wherein the amino acid nutrients include or are selected from one or more of the following amino acid compounds with nutritional health effects: amino acids, amino acid salts, oligopeptides and polypeptides; preferably amino acids or their salts selected from the group consisting of amino acids or oligopeptides and polypeptides comprising or consisting of the following amino acids: alanine, valine, leucine, isoleucine, phenylalanine, Proline, tryptophan, tyrosine, serine, cysteine, methionine, asparagine, glutamine, threonine, lysine, arginine, histidine, aspartic acid, Glutamic acid, beta-alanine, taurine, gamma aminobutyric acid (GABA), tea polyphenols (theanine), pumpkin seed amino acid (3-amino-3-carboxypyranoic acid), glutamine , citrulline, ornithine; more preferably amino acids or salts thereof selected from the group consisting of oligopeptides and polypeptides comprising or consisting of the following amino acids: arginine, lysine, glycine, cysteine, Alanine, serine, glutamic acid.
  14. 根据权利要求8-10之一或13的应用、药物组合物或方法，其中所述氨基酸类营养素包括甘氨酸。The use, pharmaceutical composition or method according to one of claims 8-10 or 13, wherein the amino acid nutrient comprises glycine.
  15. 根据权利要求8、9或11的应用、药物组合物或方法，其中所述活体染料为选自亚甲蓝类活体染料之一种或多种，其中所述亚甲蓝类活体染料包括或选自以下之一种或多种:亚甲蓝、专利蓝、异硫蓝及其衍生物。The use, pharmaceutical composition or method according to claim 8, 9 or 11, wherein the vital dye is one or more selected from methylene blue-based vital dyes, wherein the methylene blue-based vital dye includes or is selected from One or more of the following: methylene blue, patent blue, isothiocyanate and derivatives thereof.
  16. 根据权利要求6、7或10的应用、药物组合物或方法，其中所述酸消融剂为选自：弱酸、强酸。The use, pharmaceutical composition or method according to claim 6, 7 or 10, wherein the acid ablating agent is selected from the group consisting of: weak acid, strong acid.
  17. 根据权利要求16的应用、药物组合物或方法，其中The use, pharmaceutical composition or method according to claim 16, wherein
    所述酸消融剂为弱酸，且其给药浓度(w/v)为≥2.0％、≥3.0％或≥5％、优选3.0-10％或5-20％；或The acid ablating agent is a weak acid and its administration concentration (w/v) is ≥2.0%, ≥3.0% or ≥5%, preferably 3.0-10% or 5-20%; or
    所述酸消融剂为强酸，且其给药浓度(w/v)为≥0.5％、≥0.75％或≥1％、优选0.5-3％或0.5-3％。The acid ablating agent is a strong acid, and its administration concentration (w/v) is ≥0.5%, ≥0.75% or ≥1%, preferably 0.5-3% or 0.5-3%.
  18. 根据权利要求17或18的应用、药物组合物或方法，其中所述弱酸包括或选自以下之一种或多种：碳酸、乙酸、羟基乙酸、丙酸、丙二酸、丁酸、丁二酸、乳酸(2-羟基丙酸)、柠檬酸(2-羟基-1，2，3-丙三羧酸)、苹果酸(2-羟基丁二酸)、酒石酸、草酸、葡萄糖酸，优选为乙酸；所述强酸例如包括盐酸、硫酸、硝酸、高氯酸、硒酸、氢溴酸、氢碘酸，优选为盐酸。The use, pharmaceutical composition or method according to claim 17 or 18, wherein the weak acid comprises or is selected from one or more of the following: carbonic acid, acetic acid, glycolic acid, propionic acid, malonic acid, butyric acid, succinic acid acid, lactic acid (2-hydroxypropionic acid), citric acid (2-hydroxy-1,2,3-propanetricarboxylic acid), malic acid (2-hydroxysuccinic acid), tartaric acid, oxalic acid, gluconic acid, preferably Acetic acid; for example, the strong acid includes hydrochloric acid, sulfuric acid, nitric acid, perchloric acid, selenic acid, hydrobromic acid, and hydroiodic acid, preferably hydrochloric acid.
  19. 根据权利要求14或18的应用、药物组合物或方法，其中所述局部协同物包括甘氨酸和乙酸。The use, pharmaceutical composition or method according to claim 14 or 18, wherein the topical synergist comprises glycine and acetic acid.
  20. 根据权利要求8、9或15的应用、药物组合物或方法，其中所述局部协同物包括亚甲蓝和聚多糖,其中所述聚多糖包括或选自：羧甲基葡聚糖、水溶性β-D-葡聚糖、和右旋糖酐。The use, pharmaceutical composition or method according to claim 8, 9 or 15, wherein the topical synergist comprises methylene blue and a polysaccharide, wherein the polysaccharide comprises or is selected from the group consisting of: carboxymethyl dextran, water soluble Beta-D-glucan, and Dextran.
  21. 根据权利要求1-20之一的应用、药物组合物或方法，其中所述靶区组织为包含肿瘤细胞或/和成纤维细胞的局部病变组织。The use, pharmaceutical composition or method according to any one of claims 1-20, wherein the target tissue is a local diseased tissue comprising tumor cells or/and fibroblasts.
  22. 根据权利要求21的药物组合物、应用或方法，其中所述局部病变包括肿瘤、非瘤肿大、局部炎症、分泌腺功能异常和皮肤病,其中所述肿瘤包括恶性肿瘤和非恶性肿瘤。The pharmaceutical composition, use or method according to claim 21, wherein said local lesions include tumors, non-tumoral enlargements, local inflammation, abnormal secretion of glands and skin diseases, wherein said tumors include malignant tumors and non-malignant tumors.
  23. 根据权利要求22的应用、药物组合物或方法，其中所述恶性肿瘤包括淋巴瘤和实体肿瘤，如乳腺癌、胰腺癌、甲状腺癌、鼻咽癌、***癌、肝癌、肺癌、肠癌、口腔癌、食道癌、胃癌、喉癌、睾丸癌、***癌、子宫癌、卵巢癌。The use, pharmaceutical composition or method according to claim 22, wherein said malignancies include lymphomas and solid tumors, such as breast cancer, pancreatic cancer, thyroid cancer, nasopharyngeal cancer, prostate cancer, liver cancer, lung cancer, bowel cancer, oral cancer Cancer, esophageal cancer, stomach cancer, throat cancer, testicular cancer, vaginal cancer, uterine cancer, ovarian cancer.
  24. 根据权利要求22的应用、药物组合物或方法，其中所述非恶性肿瘤包括实体肿瘤，如乳腺瘤、胰腺、甲状腺瘤、***瘤、肝瘤、肺瘤、肠瘤、口腔瘤、食道瘤、胃瘤、鼻咽瘤、喉瘤、睾丸瘤、***瘤、子宫瘤、输卵管瘤、卵巢瘤。The use, pharmaceutical composition or method according to claim 22, wherein said non-malignant tumor comprises solid tumors, such as breast tumor, pancreas, thyroid tumor, prostate tumor, liver tumor, lung tumor, intestinal tumor, oral tumor, esophageal tumor, Gastric tumor, nasopharyngeal tumor, laryngeal tumor, testicular tumor, vaginal tumor, uterine tumor, fallopian tube tumor, ovarian tumor.
  25. 根据权利要求3-24之一的方法，其中所述方法包括在局部用药所述药物组合物之前、期间或之后还任选进行一种或多种其它治疗，例如化疗、免疫疗法、放射疗法、手术、物理消融。The method according to any one of claims 3-24, wherein said method comprises optionally one or more other treatments, such as chemotherapy, immunotherapy, radiation therapy, prior to, during or after topical administration of said pharmaceutical composition. Surgery, physical ablation.
  26. 一种用于治疗局部病变疾病的装置，其包含根据权利要求1-24之一的药物组合物。A device for the treatment of local pathological diseases, comprising a pharmaceutical composition according to any one of claims 1-24.
PCT/CN2022/076823 2021-02-18 2022-02-18 Pharmaceutical composition comprising immunoglobulin, and use thereof WO2022174812A1 (en)

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