Formulations containing N,N-dimethylbiguanide, and pharmaceutical combinations, compositions, kits, uses, and methods of preparation containing N,N-dimethylbiguanide and a drug.

Abstract

This disclosure relates to formulations comprising N,N-dimethylbiguanide and their use in the preparation of interventional drugs for lesions. This application also relates to pharmaceutical combinations comprising N,N-dimethylbiguanide and synergistic agents, pharmaceutical compositions comprising such pharmaceutical combinations, formulations, kits, methods of preparation thereof, and uses thereof.

Description

Technical Field

[0001] This application discloses novel uses of metformin compounds, and their advantages based on these novel uses (reduced dosage and enhanced efficacy, reduced biological barrier function, improved heterogeneity, etc.) in combination with different components (Z) and their applications. Z includes or is selected from at least one of the following: cell-responsive antitumor drugs (abbreviated as B), (injectable) weakly basic sodium salts (abbreviated as C), methylene blue dyes (abbreviated as D), and immunomodulators (abbreviated as E), such as injectable immunomodulators. Prior Technology

[0002] Many substances exhibit antitumor activity in cell experiments, but even those with strong activity often show a low probability of demonstrating the expected effect in animal experiments. Conversely, those with good activity often show a low probability of demonstrating the expected therapeutic effect in clinical trials. Only a very small number of top-performing substances are approved for indications for the treatment of the most preferred tumors. However, even after careful selection, clinical practice has found that even within the scope of indications, efficacy varies from person to person, and the efficacy is often unsatisfactory. The key limitations are as follows:

[0003] Biological barrier issues. These cell-reactive drugs or their active forms can only produce an anti-tumor response upon contact with tumor cells. Cell suspensions are too simple compared to tumor tissue in animal models, while tumor tissue in animal patients is much more complex, and the degree of biological barriers (tissue barriers, cell membrane barriers, etc.) is also significantly increased, becoming a key control step for their cellular response.

[0004] Heterogeneity is a significant issue. Heterogeneity includes tissue heterogeneity, spatial atypia, and safety heterogeneity. Different tumors of the same type, or different regions of the same tumor, can vary greatly in tissue barrier function, morphological distribution, and safety impact. If drug trials are based solely on cell or animal studies without adequately considering these heterogeneity issues, antitumor drugs, even those administered directly to tumor sites, will struggle to be administered according to individual tumor characteristics, making it difficult to achieve the desired therapeutic effect.

[0005] Dose-response issues. The efficacy of cell-responsive drugs is highly dependent on their antitumor concentration, while their dosage is often limited by their side effects. Many drugs come at the cost of reduced safety, often causing patients to suffer from side effects during treatment, and the efficacy (time-bound, effectiveness, long-lasting, and heterogeneous adaptation) provided at higher doses still needs improvement. For example, due to the superposition of biological barriers and heterogeneity, the amount of drug delivered to different regions and the duration of maintaining that amount can vary greatly. If the dose-response effect is also weak, the antitumor effect in some regions may be insufficient to effectively inhibit tumor cells, and these regions may become sources of tumor resistance and recurrence. Thus, the weak dose-response effect of the drug includes low heterogeneity even at high doses.

[0006] In existing technologies, metformin (usually C4H11N5.HCl) is indicated for lowering blood glucose levels in type 2 diabetes. Research into novel uses for metformin is ongoing, and all reported antitumor effects of metformin (hereinafter referred to as known metformin antitumor effects) are against tumor cells, such as influencing tumor cell proliferation through the activation of protein kinase pathways via AMP (Morales DR, Morris A D. Metformin in Cancer Treatment and Prevention[J]. Annual Review of Medicine, 2015.). Its half-maximal effective concentration (EC50) against selected sensitive cells in cell experiments is in the mM range, more than 100 times higher than the μM range of standard antitumor drugs such as 5-FU, indicating that even if metformin has antitumor effects at the cellular level, they are very weak.

[0007] Compared to approved drugs indicated for treating solid tumors, metformin exhibits more pronounced limitations in the three key areas mentioned above (biological barrier issues, heterogeneity issues, and dose-response issues). In summary, standard metformin drugs or formulations are characterized by high dosage and low efficacy (weak dose-response), requiring long-term (e.g., continuous administration for more than 30 days in mouse experiments) high-dose administration (e.g., 1000-2000 mg daily) to produce still very low efficacy. For example, in five metformin clinical studies using Ki-67 as a tumor proliferation marker, only two studies showed a significant decrease in Ki-67, two studies showed no significant change in Ki-67, and one study found an increase in Ki-67 levels (Vancura A, Bu P, Bhagwat M, et al. Metformin as an Anticancer Agent[J]. Trends Pharmacol. Sci., 2018., Vol. 39, No. 10, pp. 867-878).

[0008] The co-use of standard metformin with other drugs (such as cell-responsive antitumor drugs) essentially utilizes their interactions at the cellular level (Kan Guanting, Yu Jiandong, Xiong Yang, Research Progress on Metformin Combined with Chemotherapy Drugs for Antitumor Treatment, Chinese Journal of Modern Applied Pharmacy, August 2020, Vol. 37, No. 16, pp. 2025-2030). This co-use does not alter the antitumor pharmacology of each active component, the aforementioned weak dose-response of standard metformin remains, and the problems to be solved do not involve the aforementioned biological barriers and heterogeneity issues.

[0009] This application relates to local lesions, taking tumors as an example, and aims to address the aforementioned problems of standard metformin and its use with other drugs (such as cell-reactive antitumor drugs) (Problem 1: weak dose-response, especially under conditions of biological barriers and heterogeneity; Problem 2: when used with other drugs, it can only provide the cytotoxic effects limited by Problem 1, without helping to solve the biological barrier and heterogeneity problems of the latter). The goal is to endow metformin and its use with antitumor drugs with new therapeutic value (such as new indications) that are not available in the existing technology but are urgently needed in clinical practice, so as to meet various clinical needs that cannot be met by the existing technology. Summary of the Invention

[0010] The inventors of this invention unexpectedly discovered that metformin hydrochloride aqueous solution and N,N-dimethylbiguanide aqueous solution, which are usually regarded as pharmaceutical equivalents, showed differences in intratumoral cell morphology after one hour of intervention in tumors containing drug-resistant tumor cells, and differences in tumor inhibition rate at the experimental endpoint. These results clearly contradict the existing structure-activity concept that "same basic chemical structure, basically the same efficacy" (which is also the basis for the common use of metformin salt as metformin in the prior art), and exceed the expectations of the typical antitumor effect of metformin.

[0011] Therefore, according to a first aspect of the invention, a product specifically for interventional procedures is provided, specifically a formulation comprising N,N-dimethylbiguanide (abbreviated as R) and optionally a medium, such as a solvent, wherein the formulation does not contain an acid capable of converting R into its salt, such as an acidic pH adjuster required to meet the safety requirements of cell culture or standard injection, wherein the concentration of the N,N-dimethylbiguanide is ≥ its application concentration, wherein the application concentration (w / w) is 0.5% ≤ CR ≤ 51%, preferably 1%-50%, more preferably 3%-50% or 5%-30%.

[0012] More specifically, the formulations of the present invention are preferably used to treat lesions such as tumors or nodules in subjects, and preferably, the treatment is a local treatment.

[0013] In some embodiments, the tumor includes a solid tumor.

[0014] In some embodiments, the above-mentioned formulation includes a medium such as a solvent, preferably water, more preferably water for injection.

[0015] In some embodiments, the above formulation does not include a salt of N,N-dimethylbiguanide (referred to as metformin salt, or R').

[0016] In some embodiments, the concentration (w / w) of N,N-dimethylbiguanide salt in the above formulation is ≤30%, preferably ≤25%, preferably ≤10%, more preferably ≤5%, ≤4% or ≤3%, even more preferably ≤2%, particularly preferably ≤1%, such as 1% or 0.5%, and most preferably 0, that is, the formulation does not contain N,N-dimethylbiguanide salt.

[0017] In some embodiments, the amount of N,N-dimethylbiguanide used in the above formulation is: In the formula, nR represents the number of times R is applied to the tumor in one course of treatment, qR(i) represents the dosage of R in the i-th application (i=1, ..., nR), and: In the formula, nRi is the number of application points of R at the i-th application, such as the number of injection points, cR(ii) and vR(ii) are the concentration and volume of R applied at the ii-th application point, such as the injection point, respectively, where vR(ii) ≦ 3 times the target volume of application point ii (abbreviated as vtarget(ii)), and CR is 0.5-51%, where 0.5% is the common concentration threshold of R and 51% is the upper limit of the dissolution threshold of R.

[0018] In some embodiments, the formulation is a fast-acting formulation used only to rapidly reduce the size of lesions.

[0019] In some embodiments, the formulation is a low-frequency effective formulation intended only for low-frequency administration.

[0020] In some embodiments, the formulation is a liquid provider, wherein the liquid is a microvolume liquid, and the microvolume is 2-1000 μl, 5-500 μl, 5-100 μl, 5-50 μl, or 5-20 μl.

[0021] In some embodiments, the salts of N,N-dimethylbiguanide include hydrochloride, sulfate, acetate, phosphate, citrate, lactate, nitrate, carbonate, etc.

[0022] In some embodiments, the acidic pH adjuster includes or is selected from strong acids such as hydrochloric acid and sulfuric acid; or weak acids such as acetic acid, citric acid, phosphoric acid, malic acid, tartaric acid, lactic acid, and carbonic acid.

[0023] In some embodiments, N,N-dimethylbiguanide is an organic compound (a) with the chemical formula C4H11N5, having the following formula:

[0024] In some embodiments, the metformin drug used as the standard drug is typically a complex of N,N-dimethylbiguanide and an acid (referred to as metformin salt, or R'), with the chemical formula C4H11N5.acid(a+b). The most commonly used metformin drug is metformin hydrochloride (C4H11N5.HCl), i.e., metformin hydrochloride, which has the following formula 2:

[0025] In some embodiments, the formulation comprises N,N-dimethylbiguanide and water. In one embodiment, the application concentration (CR, w / w) of N,N-dimethylbiguanide in the formulation can be 0.75%, 1%, 2%, 3%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, or 50%, or any range therebetween, such as 0.75% to 20%, 1% to 20%, 2% to 20%, 3% to 20%, 5% to 20%, 5% to 40%, etc. In another embodiment, the application concentration (CR, w / w) of N,N-dimethylbiguanide in the formulation is 20%, 30%, 35%, or 40% to a maximum solubility such as 51%, or can be in the range of 0.5%-51%, 0.5-50%, 1%-50%, 5%-50%, 15%-50%, 25%-50%, 35%-50%.

[0026] In some embodiments, the formulation does not contain an acid that would cause N,N-dimethylbiguanide in the aqueous solution to be significantly converted to metformin salt. Specifically, the acid is an acidic pH adjuster required to meet the safety requirements of cell culture or standard injection, including or selected from, for example, hydrochloric acid, sulfuric acid, acetic acid, citric acid, phosphoric acid, malic acid, tartaric acid, lactic acid, and carbonic acid.

[0027] In some embodiments, the formulation is specifically designed for interventional treatment of lesions, preferably for treating lesions such as tumors or nodules in a subject.

[0028] According to a second aspect of the invention, a formulation specifically for interventional treatment of lesions is provided, comprising N,N-dimethylbiguanide (abbreviated as R) and optionally a solvent, wherein the formulation does not contain an acid capable of converting said R into its salt, such as an acidic pH adjuster required to meet the safety requirements of cell culture or standard injection, wherein the concentration of said N,N-dimethylbiguanide is ≥ its interventional concentration, wherein said interventional concentration (w / w) is 0.5% ≤ C ≤ 51%, preferably 1%-50%, more preferably 3%-40% or 5%-35%, preferably used for interventional treatment of lesions such as tumors or nodules in subjects.

[0029] In some embodiments, the formulation may contain a solvent, preferably water, and more preferably water for injection.

[0030] In some embodiments, the above formulation may not include metformin salt.

[0031] In some embodiments, the N,N-dimethylbiguanide in the above-described formulation has the structure described in the first aspect (Formula 1), and its concentration (w / w) can be ≤51%, ≤50%, ≤40%, ≤30%, ≤25%, ≤20%, or ≤10%, ≤5%, ≤2%, such as 1%, 0.75%, or 0.5%.

[0032] In some embodiments, the concentration (w / w) of N,N-dimethylbiguanide(R) in the formulation may be 0.5%-51%, 0.5-50%, 1%-40%, 5%-50%, 5%-40%, 5%-30%, 5%-25%, 5%-20%, 1%-30%, 1%-25%, 1%-20%, 15%-50%, 25%-50%, 35%-50%, 5%-20%, 0.5%-10%, or 1%-5%, or ≤2%, such as 1%, 0.75%, or 0.5%.

[0033] In some embodiments, the acidic pH adjuster may include or be selected from hydrochloric acid, sulfuric acid, acetic acid, citric acid, phosphoric acid, malic acid, tartaric acid, lactic acid, and carbonic acid.

[0034] In some embodiments, the formulation consists of N,N-dimethylbiguanide (R) and water.

[0035] In some embodiments, the formulation does not contain an acid that would cause N,N-dimethylbiguanide(R) in the aqueous solution to be significantly converted into metformin salt.

[0036] According to a third aspect of the invention, a pharmaceutical composition is provided comprising N,N-dimethylbiguanide (R) and a component (Z), wherein the N,N-dimethylbiguanide is administered at a concentration of 0.1%-51% by weight (w / w), preferably 0.2%-50%, and the component (Z) comprises or is selected from at least one of the following: a cell-responsive antitumor drug (abbreviated as B), a near-neutral or weakly basic sodium salt (abbreviated as C), a methylene blue dye (abbreviated as D), and an immunomodulator (abbreviated as E). In some embodiments, in the above-described pharmaceutical composition, N,N-dimethylbiguanide (R) has the structure described in the first aspect (Formula 1).

[0037] In some implementations, drug combinations are provided that can be used to treat lesions such as tumors or nodules in the subject.

[0038] In some embodiments, in the above-described drug combination, the N,N-dimethylbiguanide is defined by the following to provide an active form that distinguishes it from standard metformin (whose active ingredient is R'): 1) Do not mix with acids that cause R to be significantly converted to R'; 2) Application to local lesions; 3) The dosage for one course of treatment is: In the formula, QR < 50% or 25% of the equivalent dosage of R', nR is the number of times R is applied to the tumor in one course of treatment, qR(i) is the dosage of R in the i-th application (i = 1, ..., nR), and: In the formula, nRi is the number of application points of R at the i-th application, such as the number of injection points, cR(ii) and vR(ii) are the concentration and volume of R applied at the ii-th application point, such as the injection point, respectively, wherein the concentration threshold of cR(ii) (W / W) is 0.1% or 0.2%, and the upper limit of the concentration threshold is 51%, and the volume threshold of vR(ii) is 1 / 800 or 1 / 300 of the target volume (abbreviated as vtarget(ii)), and the upper limit of the volume threshold is 3 times vtarget(ii).

[0039] The active form of R' is characterized by complete complexation with acid in solution; while the active form of R is characterized by less complexation with acid in solution, mainly existing in a form of very little or partial complexation. Thus, through the transient local action of this different active form, it provides the activity that R' cannot provide (abbreviated as activity A), including at least one of the following: the effect of effectively reducing the function of biological barriers (abbreviated as activity A1), the effect of effectively damaging diseased tissue (abbreviated as activity A2), and / or the effect of effectively releasing diseased immune substances and / or inflammatory signals within the lesion (abbreviated as activity A3).

[0040] In some embodiments, the drug combination is a drug combination for treating solid tumors, comprising said R and B, wherein: - The activity A of R includes at least the effective reduction of biological barrier function (activity A1); - The cell-responsive antitumor drug (B) provides the cell response (abbreviated as active B) by means of the following, one of the key control steps of which is crossing the aforementioned biological barrier: 1) Intratumoral application and / or systemic application; 2) The dosage for one course of treatment is: In the formula, QB is the standard dosage of B in sensitive tumors, nB and qB(j) are the number of systemic administrations of B in one course of treatment and the dosage of the jth administration (j=1, ..., nB), respectively, and nB', cB(j'), and vB(j') are the number of tumor administrations of B in one course of treatment and the concentration and volume of the j'th administration (j'=1, ..., nB'), respectively.

[0041] In some embodiments, in the above-described drug combination, the drug combination includes said R, C, and / or D, wherein: - The activity A of R includes at least the effect of effectively damaging diseased tissue (activity A2); - The C is used to provide an effect that improves the local efficacy-toxicity ratio of the R (abbreviated as active C) by the following definition: 1) Apply in combination with the R; 2) The mixing ratio of C to R (QC / QR) is 0.5 to 4.0; - The D provides pharmacological efficacy that enhances the drug effect of the R by the following limitations: 1) Application to lesions, including application in combination with or separately from the R; 2) The dosage QE for one course of treatment is ≤ the effective dosage for transient local effects.

[0042] In some embodiments, the above-described drug combination comprises the R and E, wherein: - The activity A of R includes at least the function of effectively reducing the function of biological barriers (activity A1) and / or the function of effectively releasing disease immune substances and / or inflammatory signals within the lesion (activity A3); - The E is defined by the following functions for providing immunomodulatory effects benefiting from the effects of the active A1, and / or optimizing local and / or systemic immune responses by utilizing the effects of the active A3 (abbreviated as active E): 1) Intralesional application and / or systemic application; 2) The dosage for one course of treatment is: In the formula, QE is the standard dosage of E in the anti-lesion application, nE and qE(k) are the number of systemic applications of E in one course of treatment and the dosage of the kth application (j=1, ...nk), respectively, and nE', cE(k') and vE(k') are the number of tumor applications of E in one course of treatment and the concentration and volume at the k'th application (k'=1, ...nk'), respectively.

[0043] In some embodiments, in the above-described drug combination, the tumor is a solid tumor suitable for intratumoral administration. In some embodiments, in the above-described drug combination, component (Z) includes a cell-responsive antitumor drug (B), wherein R provides activities (abbreviated as activity A) that are not provided by standard metformin (the active ingredient being a metformin salt, abbreviated as R'), including effectively reducing biological barrier function (abbreviated as activity A1): 1) Application to the tumor; 2) Application rate: In the formula, nR represents the number of times R is applied to the tumor in one course of treatment, qR(i) represents the dosage of R in the i-th application (i=1, ..., nR), and: In the formula, nRi is the number of injection points for R at the i-th application, cR(ii) and vR(ii) are the concentration and volume of R applied at the ii-th injection point, respectively. The concentration threshold of cR(ii) (W / W) is 0.1% or 0.2%, and the upper limit of the concentration threshold is 51%. The volume threshold of vR(ii) is 1 / 800 or 1 / 300 of the target volume (abbreviated as vtarget(ii)), and the upper limit of the volume threshold is 3 times vtarget(ii). - The B provides one of its key control steps (abbreviated as active B) through the following definition, which enables its specific antitumor action across the tumor tissue barrier: 1) Intratumoral application and / or systemic application; 2) Application rate: In the formula, QB is the standard dosage of B in tumors sensitive to B, nB and qB(j) are the number of systemic administrations of B in one course of treatment and the dosage of the j-th administration (j=1, ..., nB), respectively, and nB', cB(j') and vB(j') are the number of tumor administrations of B in one course of treatment and the concentration and volume of the j'th administration (j'=1, ..., nB'), respectively. - The type of combination is a synergistic combination in which the drug effects of activity A of R and activity B of B mutually enhance each other.

[0044] In some embodiments, in the above-described drug combination, component (Z) includes a near-neutral or weakly basic sodium salt (C); R is limited to lesion application, and C improves the efficacy-toxicity ratio of R for lesion application by mixing with said R; and the mixing ratio of C to R (QC / QR) is 0.5 to 4.0.

[0045] In some embodiments, in the above-described drug combination, component (Z) includes the methylene blue dye (D); R and D are limited to lesion application, and D enhances the efficacy of R by mixing with R or by applying it independently.

[0046] In some embodiments, in the above-described drug combination, component (Z) includes an immunomodulator (E), wherein: -R is defined as providing activities (abbreviated as Activity A) that cannot be provided by standard metformin (the active ingredient is a metformin salt, abbreviated as R'), including effectively reducing biological barrier function (abbreviated as Activity A1), and / or effectively releasing disease immune substances and / or inflammatory signals within the lesion (abbreviated as Activity A3): 1) Application to the lesion; 2) The dosage for one course of treatment is: ,

[0047] In the formula, nR represents the number of times R is applied to the lesion in one course of treatment, qR(i) represents the dosage of R in the i-th application (i=1, ..., nR), and: In the formula, nRi is the number of application points of R at the i-th application, such as the number of injection points, cR(ii) and vR(ii) are the concentration and volume of R applied at the ii-th application point, such as the injection point, respectively, wherein the concentration threshold of cR(ii) (W / V) is 0.1% or 0.2%, and the upper limit of the concentration threshold is 51%, and the volume threshold of vR(ii) is 1 / 800 or 1 / 300 of the target volume (abbreviated as vtarget(ii)), and the upper limit of the volume threshold is 3 times vtarget(ii); - The E is defined as having the function of providing immunomodulatory effects that benefit from reduced biological barrier function, and / or optimizing local and / or systemic immune responses by utilizing diseased immune substances and / or inflammatory signals effectively released within the lesion (abbreviated as Active E): 1) Intralesional application and / or systemic application; 2) The dosage for one course of treatment is: In the formula, QE is the standard dosage of E in the anti-lesion application, nE and qE(k) are the number of systemic applications of E in one course of treatment and the dosage of the kth application (j=1, ...nk), respectively, and nE', cE(k') and vE(k') are the number of tumor applications of E in one course of treatment and the concentration and volume at the k'th application (k'=1, ...nk'), respectively.

[0048] In some implementations, the concentration threshold of cR(ii) (W / W) can be 0.1% (synergistic threshold) or 0.2% (effective threshold), and the upper limit of the concentration threshold is 51%; the volume threshold of vR(ii) can be 1 / 800 or 1 / 300 of the target volume (abbreviated as vtarget(ii)), and the upper limit of the volume threshold is 3 times vtarget(ii).

[0049] In some embodiments, in the above-described drug combination, N,N-dimethylbiguanide (R) comprises <30% (w / w), preferably ≤5% metformin salt (R'), or more preferably excluding metformin salt (R'). Preferably, N,N-dimethylbiguanide (R) is in the form of a powder for injection.

[0050] In some embodiments, the above-described drug combination is applicable to local lesions such as tumors. Preferably, the drug combination includes B and / or E for refractory malignant tumors. Preferably, the refractory malignant tumor includes or is selected from at least one of the following groups: chemotherapy-resistant tumors, tumors with unfavorable microenvironments, tumors that have been discontinued from antitumor drugs, tumors for which there are no effective chemotherapy drugs, and tumors carried by patients with contraindications or inappropriate conditions for standard chemotherapy drugs.

[0051] In some embodiments, the above-described drug combination is applicable to heterogeneous tumors, wherein the heterogeneity includes at least one of tissue heterogeneity, spatial heterogeneity, and safety heterogeneity, and the heterogeneous tumor is a tumor in which differentiated treatment is beneficial by setting different tumors of the same type or different regions of the same tumor as different target areas based on the heterogeneity.

[0052] In some embodiments, in the above-described drug combination, component (Z) includes an antitumor drug, which includes or is selected from cell-reactive drugs (B) whose key control step is to cross the tumor tissue barrier.

[0053] In some embodiments, the cell-responsive drugs in the above-described drug combination include cytotoxic drugs and targeted antitumor drugs.

[0054] In some embodiments, the cytotoxic drugs in the above-described drug combination include DNA damaging agents, antimetabolites, microtubule inhibitors, and topoisomerase inhibitors.

[0055] In some embodiments, representative compounds of DNA-damaging agents in the above-described drug combination include cisplatin, carboplatin, oxaliplatin, ifosfamide, and doxorubicin.

[0056] In some embodiments, representative compounds of antimetabolites in the above-described drug combination include fluorouracil, gemcitabine, and methotrexate.

[0057] In some embodiments, representative compounds of the microtubule inhibitors in the above-described drug combination include paclitaxel and vincristine.

[0058] In some embodiments, the targeted antitumor drugs in the above-described drug combination include gefitinib, osimertinib, larotrectinib, pazopanib, trastuzumab and their derivatives.

[0059] In some embodiments, in the above-described drug combination, drug B includes or is selected from at least one of the following: cisplatin, cyclophosphamide, carboplatin, doxorubicin, ifosfamide, nitrogen mustard, fluorouracil, capecitabine, gemcitabine, methotrexate, pemetrexed, fludarabine, paclitaxel, docetaxel, vincristine, vinca, vindesin, etoposide, irinotecan, topotecan, daunorubicin, mitoxantrone, gefitinib, osimertinib, larotrectinib, pazopanib, trastuzumab and its derivatives.

[0060] In some embodiments, in the above-described drug combination, the near-neutral or weakly basic sodium salt (C) includes or is selected from one or more of the following: sodium chloride, sodium bicarbonate, sodium dihydrogen phosphate, sodium lactate, sodium acetate, etc. Preferably, the weakly basic sodium salt (C) includes an injectable weakly basic sodium salt, such as sodium bicarbonate.

[0061] In some embodiments, in the above-described drug combination, the methylene blue dye (D) includes or is selected from one or more of the following: methylene blue, patent blue, isothiocyanate, and neomethylene blue.

[0062] In some embodiments, in the above-described drug combination, the immunomodulator (E) includes immunomodulators and / or immunosuppressants. Specifically, the immunomodulator may include or be selected from one or more of the following: biological macromolecular immunomodulators (abbreviated as E1), immune cell immunomodulators (abbreviated as E2), and vaccine-type immunomodulators (abbreviated as E3).

[0063] In some embodiments, the biomolecular immunomodulator may include, for example, immune checkpoint inhibitors, cytokines, and TLR agonists. In one embodiment, the immune checkpoint inhibitor may include Anti-PD-1 (CD279), the cytokine may include interleukin-15, and the TLR agonist may include CpG ODN 1826 (abbreviated as CpG).

[0064] In some implementations, immune cell-based immunomodulators include, for example, CAR-T cell therapy, DC cell therapy, TIL cell therapy, etc.

[0065] In some implementations, vaccine-type immunomodulators include BCG.

[0066] In some embodiments, the application concentration (w / w) of dimethyl biguanide (R) in the drug combination can be 0.5%-51%, 0.5-50%, 0.5-45%, 1%-45%, 5%-45%, 5%-40%, 5%-30%, 5%-25%, 5%-20%, 1%-30%, 1%-25%, 1%-20%, 0.5%-10%, 1%-5%, 15%-50%, 25%-50%, 35%-50%, 5%-20%, or ≤2%, such as 1%, 0.75%, or 0.5%.

[0067] In some embodiments, the above-mentioned drug combination includes dimethyl biguanide (R) and doxorubicin (DOX), wherein the dimethyl biguanide (R) can be administered at a concentration of 0.2%-50%, and the doxorubicin (DOX) can be administered at a concentration of 0.025%-1%. Preferably, the dimethyl biguanide (R) is administered at a concentration of 0.5%-50%, and the doxorubicin (DOX) is administered at a concentration of 0.1%-1%. Optionally, both dimethyl biguanide (R) and doxorubicin (DOX) are administered to the tumor.

[0068] In some embodiments, the drug combination includes dimethylbiguanide(R) and ifosfamide (IFO), wherein the dimethylbiguanide(R) can be administered at a concentration of 0.2%-50%, and the ifosfamide (IFO) can be administered at a concentration of 0.1%-15%. Preferably, the dimethylbiguanide(R) is administered at a concentration of 0.2%-45%, and the ifosfamide (IFO) is administered at a concentration of 0.1%-10%. More preferably, the dimethylbiguanide(R) is administered at a concentration of 5%-45%, and the ifosfamide (IFO) is administered at a concentration of 0.5%-10%.

[0069] In some embodiments, the drug combination includes dimethylbiguanide (R) and cisplatin (DDP), wherein the dimethylbiguanide (R) may be administered at a concentration of 0.2%-50%, and the cisplatin (DDP) may be administered at a concentration of 0.01%-0.4%. Preferably, the dimethylbiguanide (R) is administered at a concentration of 0.5%-50%, and the cisplatin (DDP) is administered at a concentration of 0.01%-0.04%.

[0070] In some embodiments, the drug combination includes dimethylbiguanide (R) and 5-fluorouracil (5-FU), wherein the dimethylbiguanide (R) can be administered at a concentration of 0.2%-50%, and the 5-fluorouracil (5-FU) can be administered at a concentration of 0.05%-9%. Preferably, the dimethylbiguanide (R) is administered at a concentration of 0.2%-45%, and the 5-fluorouracil (5-FU) is administered at a concentration of 0.5%-9%. More preferably, the dimethylbiguanide (R) is administered at a concentration of 5%-45%, and the 5-fluorouracil (5-FU) is administered at a concentration of 0.9%-9%.

[0071] In some embodiments, the drug combination includes dimethyl biguanide (R), cisplatin (DDP), and gemcitabine (GEM), wherein the dimethyl biguanide (R) may be administered at a concentration of 0.2%-50%, the cisplatin (DDP) may be administered at a concentration of 0.05%-1%, and the gemcitabine (GEM) may be administered at a concentration of 0.5%-2%, preferably at a concentration of 1%.

[0072] In some embodiments, the drug combination includes dimethyl biguanide (R), paclitaxel (PTX), and 5-fluorouracil (5-FU), wherein the dimethyl biguanide (R) can be administered at a concentration of 0.2%-50%, the 5-fluorouracil (5-FU) can be administered at a concentration of 0.1%-10%, and the paclitaxel can be administered at a concentration of 0.02-1%, preferably at a concentration of 0.03%.

[0073] In some embodiments, the drug combination includes bimethyl biguanide (R) and osimertinib, wherein the bimethyl biguanide (R) may be administered at a concentration of 0.2%-50%, and the osimertinib may be administered at a concentration of 0.05%-2%. Preferably, the bimethyl biguanide (R) is administered at a concentration of 5%, and the osimertinib is administered at a concentration of 0.15%.

[0074] In some embodiments, the drug combination includes dimethylbiguanide (R) and gefitinib (GEF), wherein the dimethylbiguanide (R) may be administered at a concentration of 0.2%-50%, and the gefitinib (GR) may be administered at a concentration of 0.1%-2%. Preferably, the dimethylbiguanide (R) is administered at a concentration of 1%-50%, and the gefitinib (GR) is administered at a concentration of 0.5%.

[0075] In some embodiments, the drug combination includes dimethylbiguanide (R), cisplatin (DDP), and gefitinib (GR), wherein the dimethylbiguanide (R) is administered at a concentration of 5%, the gefitinib (GR) is administered at a concentration of 0.5%, and the cisplatin (DDP) is administered at a concentration of 0.15%.

[0076] In some embodiments, the drug combination includes dimethylbiguanide (R), ifosfamide (IFO), and gefitinib (GR), wherein the dimethylbiguanide (R) is administered at a concentration of 5%, the ifosfamide (IFO) is administered at a concentration of 1%, and the gefitinib (GR) is administered at a concentration of 0.5%.

[0077] In some embodiments, the drug combination includes dimethyl biguanide (R), doxorubicin (DOX), and gefitinib (GR), wherein the dimethyl biguanide (R) is administered at a concentration of 5%, the doxorubicin (DOX) is administered at a concentration of 0.05%, and the gefitinib (GR) is administered at a concentration of 0.5%.

[0078] In some embodiments, the drug combination comprises dimethyl biguanide (R) and sodium bicarbonate (SB), wherein dimethyl biguanide (R) may be administered at a concentration of 0.5%-4%, and sodium bicarbonate (SB) may be administered at a concentration of 0.5%-15%, preferably 0.5%-10%, more preferably 1.5%-9%; preferably dimethyl biguanide (R) is administered at a concentration of 1%-3%, and sodium bicarbonate (SB) is administered at a concentration of 1%-9%; more preferably, dimethyl biguanide (R) is administered at a concentration of 3%, and sodium bicarbonate (SB) is administered at a concentration of 3%-9%. Optionally, the drug combination further comprises ifosfamide (IFO) at a concentration of 0.5%, 5-fluorouracil (5-FU) at a concentration of 0.4%, or osimertinib at a concentration of 0.3%.

[0079] In some embodiments, the drug combination includes dimethylbiguanide (R) and a methylene blue dye (D) such as methylene blue (MB). In one embodiment, the drug combination includes dimethylbiguanide (R) and methylene blue (MB), wherein the dimethylbiguanide (R) can be administered at a concentration of 1%-50%, and the MB can be administered at a concentration of 0.3%-5%, preferably 0.5%-5%; more preferably, the dimethylbiguanide (R) is administered at a concentration of 5%-50%, and the MB is administered at a concentration of 0.5%-5%; preferably, the dimethylbiguanide (R) is administered at a concentration of 5%-50%, and the MB is administered at a concentration of 0.5%-1%. Optionally, the drug combination further includes one or more of ifosfamide (IFO) at a concentration of 0.5%-1%, 5-fluorouracil (5-FU) at a concentration of 0.4%-1%, and osimertinib at a concentration of 0.3%. Preferably, the drug combination comprising bimethyl biguanide (R) and methylene blue (MB) can be administered intratumorally and / or systemically. In some embodiments, the drug combination comprises bimethyl biguanide (R), MB, and any one of IFO, 5-FU, and osimertinib, wherein bimethyl biguanide (R) is administered at a concentration of 5%, MB at a concentration of 1%, IFO at a concentration of 0.5%, 5-FU at a concentration of 0.4%, and osimertinib at a concentration of 0.3%. In some embodiments, the drug combination comprises bimethyl biguanide (R), SB, MB, and any one of IFO, 5-FU, and osimertinib, wherein bimethyl biguanide (R) is administered at a concentration of 5%, SB at a concentration of 5%-10%, MB at a concentration of 0.5%, IFO at a concentration of 0.5%, 5-FU at a concentration of 0.4%, and osimertinib at a concentration of 0.3%.

[0080] In some embodiments, the drug combination includes dimethyl biguanide (R) and an immunomodulator (E), wherein the immunomodulator includes or is selected from macromolecular immunomodulators (E1), cellular immunomodulators (E2), and vaccine-type immunomodulators (E3). In some embodiments, the macromolecular immunomodulator may be, for example, an immune checkpoint inhibitor, a cytokine, and a TLR agonist. In one embodiment, the immune checkpoint inhibitor may be Anti-PD-1 (CD279), the cytokine may include interleukin-15, and the TLR agonist may include CpG ODN 1826 (CpG), and the vaccine-type immunomodulator may be BCG.

[0081] In some embodiments, the drug combination comprises dimethylbiguanide(R) and CpG ODN 1826 (CpG), wherein dimethylbiguanide(R) may be administered at a concentration of 1%-50%, and CpG may be administered at a concentration of 0.1%; preferably, dimethylbiguanide(R) is administered at a concentration of 2.5%-50%, and CpG is administered at a concentration of 0.1%. Preferably, the drug combination further comprises MB at a concentration of 1%, IFO at a concentration of 0.5%, and / or SB at a concentration of 10%. In some embodiments, the drug combination comprises dimethylbiguanide(R) and CpG and any one or two of the following: IFO, MB, and SB, wherein dimethylbiguanide(R) is administered at a concentration of 2.5%-50%, IFO at a concentration of 0.5%, CpG at a concentration of 0.1%, MB at a concentration of 1%, and SB at a concentration of 10%.

[0082] In some embodiments, the drug combination includes bimethyl biguanide (R) and an immune checkpoint inhibitor, Anti-PD-1 (CD279), such as RMP1-14, wherein bimethyl biguanide (R) may be administered at a concentration of 0.2%-50% and RMP1-14 may be administered at a concentration of 0.1%; preferably, bimethyl biguanide (R) is administered at a concentration of 1%-50% and RMP1-14 is administered at a concentration of 0.1%.

[0083] In some embodiments, the drug combination comprises dimethylbiguanide(R) and BCG, wherein dimethylbiguanide(R) may be administered at a concentration of 1%-50%, preferably 2.5%-50%, more preferably 5%-50%, and BCG may be administered at an amount of 1×10⁶ CFU; preferably, dimethylbiguanide(R) is administered at a concentration of 50%, and BCG is administered at an amount of 1×10⁶ CFU. Optionally, the drug combination further comprises either ifosfamide (IFO) at a concentration of 0.5% or SB at a concentration of 10%. In some embodiments, the drug combination comprises dimethylbiguanide(R) and BCG and either IFO or SB, wherein dimethylbiguanide(R) is administered at a concentration of 5%-50%, BCG is administered at an amount of 1×10⁶ CFU, IFO is administered at a concentration of 0.5%, and SB is administered at a concentration of 10%.

[0084] In some embodiments, the dimethyl biguanide (R) and component (Z) in the drug combination may be administered simultaneously, sequentially, or at intervals.

[0085] In some embodiments, both dimethyl biguanide (R) and component (Z) in the drug combination are administered to the tumor (combination relationship I); R and Z are administered to the tumor and systemically, respectively (combination relationship II); or R and a portion of Z are administered to the tumor, and another portion of Z is administered systemically (combination relationship III).

[0086] In some embodiments, the drug combination can be used to treat lesions such as tumors or nodules in the subject.

[0087] According to a fourth aspect of the invention, a pharmaceutical combination is provided comprising formulation I and formulation II, wherein formulation I comprises N,N-dimethylbiguanide (R) or R and a fractional component (Z), and formulation II comprises Z but does not contain R, wherein: 1) In formulation I, the N,N-dimethylbiguanide (R) comprises ≤5% or does not comprise a salt of N,N-dimethylbiguanide; 2) The method of sharing formulation I and formulation II is as follows: Formulation I is for intratumoral administration only and cannot be used for systemic administration. Formulation II can be used for intratumoral and / or systemic administration; 3) The combined amount of Formulation I and Formulation II is limited by the combined amount of Z and R: (1). The application rate of R is: In the formula, nR represents the number of times R is applied to the tumor in one course of treatment, qR(i) represents the dosage of R in the i-th application (i=1, ..., nR), and: In the formula, nRi is the number of application points of R at the i-th application, such as the number of injection points; cR(ii) and vR(ii) are the concentration and volume of R applied at the ii-th application point, such as the injection point, respectively, where vR(ii) ≤ 3 times the target volume of application point ii (abbreviated as vtarget(ii)); and cR is 0.2-51%, where 0.2% is the common concentration threshold of R and 51% is the upper limit of the dissolution threshold of R. (2). The application rate of Z is: In the formula, QZ is the standard dosage of ≦Z, nZ and qZ(j) are the number of systemic applications of Z in one course of treatment and the dosage of the jth application (j=1, ..., nB), respectively, and nZ', cZ(j') and vZ(j') are the number of tumor applications of Z in one course of treatment and the concentration and volume of the j'th application (j'=1, ..., nZ'), respectively.

[0088] In some embodiments, in the above-described drug combination, component Z comprises or is selected from at least one of the following: cisplatin, cyclophosphamide, carboplatin, doxorubicin, ifosfamide, nitrogen mustard, fluorouracil, capecitabine, gemcitabine, methotrexate, pemetrexed, fludarabine, paclitaxel, docetaxel, vincristine, vinca, vindesin, etoposide, irinotecan, topotecan, daunorubicin, and mitoxantrone.

[0089] In some embodiments, in the above drug combinations, the contraindication for mixing R is any one of the following groups: 1) A salt of N,N-dimethylbiguanide (R'); 2) An acidifying agent that significantly converts R into R', wherein the acidifying agent includes acidic pH adjusters and other acidic substances; 3) Standard metformin drug excipients with a dosage ratio (Q excipient / QR) >3.0 or >4.0, including their standard sustained-release carriers, such as gel carriers and nanocarriers; 4) Adjuvants that are unstable in strongly alkaline aqueous solutions, including, for example, reducing sugars, cellulose, liposomes, carbomer, and polyethylene glycol-modified nanocarriers.

[0090] The aforementioned mixing taboo refers to the need to avoid mixing with it throughout the entire process of formulation, preparation, product manufacturing, and use.

[0091] In some embodiments, the sharing method in the above-described drug combination includes: 1) R and Z are mixed for intralesional or intratumoral application. 2) R and Z are used for intralesional or intratumoral administration, respectively. 3) R and Z are used for intralesional or intratumoral administration and systemic administration, respectively. 4) R and some Z are mixed or used separately for intralesional or intratumoral administration, while other Z are used for systemic administration.

[0092] In some embodiments, the shared threshold in the above drug combination is 0.5%, 1%, 3.3%, 5% or 10%.

[0093] According to a fifth aspect of the invention, a pharmaceutical composition is provided comprising N,N-dimethylbiguanide (R) or any of the above-described pharmaceutical combinations of the invention.

[0094] According to a sixth aspect of the invention, there is provided an formulation comprising any of the above-described pharmaceutical combinations or compositions according to the invention.

[0095] According to a seventh aspect of the present invention, the use of N,N-dimethylbiguanide (R) and antitumor drugs in the preparation of medicaments, drug combinations or kits for treating tumors is provided.

[0096] According to an eighth aspect of the invention, the use of the above-described formulations, pharmaceutical combinations, or compositions of the invention in the preparation of a medicament, pharmaceutical combination, or kit for treating lesions such as tumors or nodules in a subject is provided.

[0097] Specifically, the above-described formulations, drug combinations, or compositions of the present invention can be used to treat lesions such as tumors or nodules in subjects.

[0098] In the formulations, pharmaceutical combinations, or compositions of the present invention, R is used to provide transient activity, wherein the transient activity refers to the local activity provided during a transient period before the active form of R is converted to the active form of R'.

[0099] In some embodiments, in the above-described uses, the application concentration (w / w) of the dimethyl biguanide is ≤51%, ≤50%, ≤40%, ≤30%, ≤25%, ≤20%, ≤10%, ≤5%, or ≤2%, such as 1%, 0.75%, or 0.5%.

[0100] In some embodiments, in the above-described uses, the application concentration (w / w) of the N,N-dimethylbiguanide(R) is 0.2%-51%, 0.5-51%, 0.5-50%, 1%-40%, 5%-50%, 5%-40%, 5%-30%, 5%-25%, 5%-20%, 1%-30%, 1%-25%, 1%-20%, 0.5%-10%, 1%-5%, or ≤2%, such as 1%, 0.75%, or 0.5%.

[0101] In some embodiments, in the above-described uses, the concentration (w / w) of the N,N-dimethylbiguanide salt contained in the formulation, pharmaceutical combination or composition is ≤30%, preferably ≤20%, preferably ≤10%, more preferably ≤5%, ≤4%, or ≤3%, more preferably ≤2%, particularly preferably ≤1%, such as 1% or 0.5%, and most preferably 0%, that is, the formulation, pharmaceutical combination or composition does not contain the N,N-dimethylbiguanide salt.

[0102] In some embodiments, the formulation of the present invention comprises N,N-dimethylbiguanide and optionally a solvent, the formulation not containing any additives required to meet the safety requirements for cell culture or standard injection, the concentration of N,N-dimethylbiguanide in the formulation being ≥ its interventional concentration, wherein the administration or interventional concentration (w / w) is 0.5% ≤ C ≤ 51%, preferably 1-51%, 2-50%, 3-50%, 4-50%, 5-50%, 6-50%, 7-50%, 8-50%, 9-50%, 10-50%, 15-50%, 20-50%, 25-50%, 30-50%, 35-50%, 40-50%, 1-40%, 5-50%, 10-40%, 15-40%, 20-40%, 25-40%, 30-40%, 35-40%, or 1-35%, 5%-30% %, etc., or any value and range thereof, such as 1.5%, 2%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, 16.5%, 17%, 17.5%, 18%, 18.5%, 19%, 19.5%, 20%, 21%, 21.5%, 22%, 22.5%, 23%, 23.5%, 24%, 24.5%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 45%, 50%.

[0103] In some embodiments, the formulation comprises a solvent, such as a pharmaceutically acceptable carrier, preferably water, and more preferably (sterile) water for injection.

[0104] In some embodiments, the formulation does not include a salt of N,N-dimethylbiguanide.

[0105] In some embodiments, the concentration (w / w) of the N,N-dimethylbiguanide salt contained in the formulation is ≤30%, ≤25%, preferably ≤10%, preferably ≤5%, more preferably ≤4%, more preferably ≤3%, even more preferably ≤2%, particularly preferably ≤1%, and most preferably 0, that is, the formulation does not contain the N,N-dimethylbiguanide salt.

[0106] In one embodiment, the additive includes an acidic pH adjuster.

[0107] In one embodiment, the pH adjuster includes or is selected from hydrochloric acid, sulfuric acid, acetic acid, citric acid, phosphoric acid, malic acid, tartaric acid, lactic acid, carbonic acid, etc.

[0108] In some embodiments, the pharmaceutical combination or composition used above includes an antitumor drug or agent, said antitumor drug or agent including or selected from at least one of the following: cisplatin, cyclophosphamide, carboplatin, doxorubicin, ifosfamide, nitrogen mustard, fluorouracil, capecitabine, gemcitabine, methotrexate, pemetrexed, fludarabine, paclitaxel, docetaxel, vincristine, vinca, vindesin, etoposide, irinotecan, topotecan, daunorubicin, and mitoxantrone.

[0109] In some embodiments, in the above-described uses, the antitumor drug is doxorubicin (DOX), wherein the dimethyl biguanide (R) can be administered at a concentration of 0.2%-50% (w / w), and the doxorubicin (DOX) can be administered at a concentration of 0.025%-1% (w / w). Preferably, the dimethyl biguanide (R) is administered at a concentration of 5%-50% (w / w), and the doxorubicin (DOX) is administered at a concentration of 0.1%-1% (w / w).

[0110] In some embodiments, in the above-described uses, the antitumor drug is ifosfamide (IFO), wherein the dimethyl biguanide (R) can be applied at a concentration of 0.2%-50% (w / w), and the ifosfamide (IFO) can be applied at a concentration of 0.1%-10% (w / w). Preferably, the dimethyl biguanide (R) is applied at a concentration of 0.2%-45% (w / w), and the ifosfamide (IFO) is applied at a concentration of 0.1%-10% (w / w). More preferably, the dimethyl biguanide (R) is applied at a concentration of 5%-45% (w / w), and the ifosfamide (IFO) is applied at a concentration of 0.5%-10% (w / w).

[0111] In some embodiments, in the above-described uses, the antitumor drug is cisplatin (DDP), wherein the dimethyl biguanide (R) can be administered at a concentration (w / w) of 0.2%-50%, and the cisplatin (DDP) can be administered at a concentration (w / w) of 0.01%-0.4%. Preferably, the dimethyl biguanide (R) is administered at a concentration (w / w) of 0.5%-50%, and the cisplatin (DDP) is administered at a concentration (w / w) of 0.01%-0.04%.

[0112] In some embodiments, in the above-described uses, the antitumor drug is 5-fluorouracil (5-FU), wherein the dimethyl biguanide (R) can be administered at a concentration of 0.2%-50%, and the 5-fluorouracil (5-FU) can be administered at a concentration (w / w) of 0.05%-9%. Preferably, the dimethyl biguanide (R) is administered at a concentration (w / w) of 0.2%-45%, and the 5-fluorouracil (5-FU) is administered at a concentration (w / w) of 0.5%-9%. More preferably, the dimethyl biguanide (R) is administered at a concentration (w / w) of 5%-45%, and the 5-fluorouracil (5-FU) is administered at a concentration (w / w) of 0.9%-9%.

[0113] In some embodiments, in the above-described uses, the antitumor drug includes cisplatin (DDP) and gemcitabine (GEM), wherein the dimethyl biguanide (R) can be administered at a concentration (w / w) of 0.2%-50%, the cisplatin (DDP) can be administered at a concentration (w / w) of 0.05%-1%, and the gemcitabine (GEM) can be administered at a concentration (w / w) of 0.5-2%, preferably at a concentration (w / w).

[0114] In some embodiments, in the above-described uses, the antitumor drug includes paclitaxel (PTX) and 5-fluorouracil (5-FU), wherein the dimethyl biguanide (R) can be administered at a concentration of 0.2%-50% (w / w), the 5-fluorouracil (5-FU) can be administered at a concentration of 0.1%-10% (w / w), and the paclitaxel can be administered at a concentration of 0.02-1% (w / w), preferably at a concentration of 0.03% (w / w).

[0115] In some embodiments, in the above-described uses, the antitumor drug is osimertinib, wherein the dimethyl biguanide (R) can be administered at a concentration (w / w) of 0.2%-50%, and the osimertinib can be administered at a concentration (w / w) of 0.05%-2%. Preferably, the dimethyl biguanide (R) is administered at a concentration (w / w) of 5%, and the osimertinib is administered at a concentration (w / w) of 0.15%.

[0116] In some embodiments, in the above-described uses, the antitumor drug is gefitinib (GR), wherein the dimethyl biguanide (R) can be administered at a concentration (w / w) of 0.2%-50%, the gefitinib (GR) can be administered at a concentration (w / w) of 0.1%-2%, preferably, the dimethyl biguanide (R) is administered at a concentration (w / w) of 1%-50%, and the gefitinib (GR) is administered at a concentration (w / w) of 0.5%.

[0117] In some embodiments, in the above-described uses, the antitumor drug comprises cisplatin (DDP) and gefitinib (GR), wherein the dimethyl biguanide (R) is administered at a concentration of 5% (w / w), the gefitinib (GR) is administered at a concentration of 0.5% (w / w), and the cisplatin (DDP) is administered at a concentration of 0.15% (w / w).

[0118] In some embodiments, in the above-described uses, the antitumor drug comprises ifosfamide (IFO) and gefitinib (GR), wherein the dimethyl biguanide (R) is administered at a concentration of 5% (w / w), the ifosfamide (IFO) is administered at a concentration of 1% (w / w), and the gefitinib (GR) is administered at a concentration of 0.5% (w / w).

[0119] In some embodiments, in the above-described uses, the antitumor drug includes doxorubicin (DOX) and gefitinib (GR), wherein the dimethyl biguanide (R) is administered at a concentration of 5% (w / w), the doxorubicin (DOX) is administered at a concentration of 0.05% (w / w), and the gefitinib (GR) is administered at a concentration of 0.5% (w / w).

[0120] In some embodiments, in the above-described uses, dimethyl biguanide and the antitumor drug can be administered simultaneously, sequentially, or at intervals.

[0121] In some embodiments, in the above-described uses, both dimethyl biguanide and the antitumor drug are administered intralesionally or intratumorally (combination relationship I); R and B are administered intralesionally or intratumorally and systemically, respectively (combination relationship II); or R and part of B are administered intralesionally or intratumorally, and another part of B is administered systemically (combination relationship III).

[0122] In some embodiments, in the above-described uses, the N,N-dimethylbiguanide is used to provide a morphological expression distinct from that of standard metformin (whose active ingredient is R'), wherein the active form of R' is a form in which it is completely complexed with acid in solution; while the active form of R is a form in which it is less complexed with acid in solution, mainly existing in a form with very little or partial complexation, thereby providing an activity (abbreviated as activity A) that is difficult for R' to provide through the transient local action of this distinct active form, including the effect of effectively reducing the function of biological barriers (abbreviated as activity A1), the effect of effectively damaging diseased tissue (abbreviated as activity A2), and / or the effect of effectively releasing diseased immune substances and / or inflammatory signals within the lesion (abbreviated as activity A3).

[0123] In some embodiments, N,N-dimethylbiguanide is used to provide an effect that maximizes the efficacy of the antitumor drug, including the effect of effectively reducing the tumor tissue barrier function (abbreviated as Activity A).

[0124] In some embodiments, the antitumor drug is used to provide an antitumor effect that maximizes the transient effect of the N,N-dimethylbiguanide, including an antitumor effect induced at the tumor cell level (abbreviated as Activity B).

[0125] In some embodiments, the N,N-dimethylbiguanide (R) must not be mixed with a sufficient amount of R' or an acidifying agent capable of converting the R into a sufficient amount of R', wherein a sufficient amount means an amount such that the ratio of the amount of R' to R (R' / R) is ≥1 / 3.

[0126] In some embodiments, in the above-described uses, a synergistic effect occurs between N,N-dimethylbiguanide and the antitumor drug, which is a dose-reduction synergistic effect relative to the standard co-occurrence combination (R' / B combination), manifested as providing the following better effect even when the metformin dosage is at least halved (QR ≦ 1 / 2QR') and the B dosage is at least reduced by 20%: 1) Significantly improved drug performance, including adaptability to tumor heterogeneity, manifested by an improvement of at least one drug adjustability index of at least 100%, wherein drug adjustability refers to the ability of the drug to provide differentiated administration parameters to different target areas while maintaining efficacy; and / or 2) The efficacy of the drug is significantly enhanced, as evidenced by an increase of at least 25% in one of the drug efficacy indicators.

[0127] In some embodiments, in the above-described uses, the synergy further constitutes a further enhancement relative to the strongest monotherapy (single drug) regimen among R and B, manifested as an increase of at least 10% in at least one of the drug adjustability indicators or / and an enhancement of at least 10% in at least one of the drug efficacy indicators.

[0128] In some embodiments, the drug adjustability index includes at least one of the following tumor administration adjustability coefficients: 1) A concentration-adjustable coefficient of R, which is the ratio of the highest value of cR to the threshold (highest value of cR / threshold), wherein the increase in the adjustable index is the increase in the coefficient, preferably the coefficient is increased to >10, 50, 100 or 150, which allows R to provide highly variable concentrations to different regions while maintaining its stable activity, including, for example, individualized drug administration for heterogeneous tumors; 2) The target volume adjustable coefficient of R is the ratio of the highest value of the target volume (v_target) to the threshold (highest value of v_target / threshold), and the increase of the adjustable index is the increase of the coefficient, preferably the coefficient is increased to >2, 10, 50, or 100, which allows R to select different v_targets for different treatment situations, including, for example, individualized drug administration for heterogeneous tumors; 3) The adjustable volume ratio coefficient of R, which is the ratio of the highest value of the ratio of the administered volume (vR) to its target volume (vtarget) to a threshold (vR / vtarget maximum value / threshold), wherein the increase in the adjustable index is the increase in the coefficient, preferably the coefficient is increased to >5, 10, or 100, which allows R to provide a highly variable volume ratio to different regions while still maintaining its stable activity, including, for example, individualized drug delivery for heterogeneous tumors; 4) The single-dose adjustment coefficient of R, which is the ratio of the highest value of the single-dose (cRi×vRi) to the threshold (highest value of cRi×vRi / threshold), wherein the increase in the adjustable index is the increase in the coefficient, preferably the coefficient is increased to >2, 6, or 10, which allows R to be selected with different single-dose doses for different treatment situations, including, for example, individualized treatment for heterogeneous tumors; and 5) The dosing point density adjustable coefficient of R is the ratio of the highest value of the dosing point density (nii / v target) to the threshold (highest value of nii / v target / threshold), and wherein the increase in the adjustable index is the increase in the coefficient, preferably the coefficient is increased to >3, 10, 50, or 100, which allows R to provide highly variable densities to different regions while maintaining its activity stability, including, for example, individualized dosing for heterogeneous tumors.

[0129] In some embodiments, the drug efficacy indicator includes at least one of the following: 1) Timeliness indicator, expressed as the time to significant effect in mouse trials (the day when tumor proliferation rate is <42%), and the time to significant effect is reduced by at least 25% compared to the R' regimen, the R' / B combined regimen, and the B monotherapy regimen, which makes the combination a rapid-acting or even more rapid-acting combination, especially suitable for tumors that require rapid reduction in tumor size; 2) Efficacy indicators, expressed as tumor inhibition rate or objective response rate in mouse trials, wherein the tumor inhibition rate or objective response rate is at least 25% higher than the control regimen (the R' regimen, the regimen in which R' is used in combination with B, D, and / or E, or the regimen in which B, D, and / or E are monotherapy), which makes the combination a highly effective or even more highly effective combination, especially suitable for tumors that require maximum inhibition of tumor size; 3) Long-acting indicators, expressed as progression-free time or survival rate in mouse trials, wherein the progression-free time or survival rate is improved by at least 25% compared to the control regimen (the R' regimen, the regimen of R' combined with B, D, or / and E, and the regimen of B, D, or / and E monotherapy), which makes the combination a long-acting or further long-acting combination, especially suitable for tumors in which it is necessary to prolong the progression-free time as much as possible.

[0130] According to a ninth aspect of the invention, a method for treating cells of a lesion, such as a tumor, is provided, comprising applying or intervening the above-described preparations, pharmaceutical combinations, or compositions of the invention into the lesion, penetrating the lesion tissue, and contacting the cells within the tissue.

[0131] According to a tenth aspect of the present invention, a method for treating a tumor is provided, comprising: administering the above-described preparations, drug combinations, and compositions of the present invention to the tumor, preferably, the administration comprising administering the preparations of the present invention to the tumor, and / or administering the metformin to the tumor and administering the drug intratumorally or systemically.

[0132] According to an eleventh aspect of the present invention, a method for treating a tumor in a subject is provided, comprising administering the above-described preparations or drug combinations of the present invention to the subject.

[0133] In the above method, the formulation comprises dimethyl biguanide and optionally a solvent, wherein the formulation does not contain an acidic pH adjuster required to meet the safety requirements of cell culture or standard injection, wherein the concentration of N,N-dimethyl biguanide is ≥ its intervention concentration, wherein the intervention concentration (w / w) is 0.5% ≤ C ≤ 51%, preferably 1%-50%, more preferably 3%-40% or 5%-35%.

[0134] In some embodiments, the formulation may contain a solvent, preferably water, more preferably water for injection.

[0135] In some embodiments, the formulation may not include metformin salt.

[0136] In some embodiments, the concentration (w / w) of the dimethyl biguanide in the formulation can be ≤51%, ≤50%, ≤40%, ≤30%, ≤25%, ≤20%, ≤10%, ≤5%, or ≤2%, such as 1%, 0.75%, or 0.5%.

[0137] In some embodiments, the concentration (w / w) of N,N-dimethylbiguanide(R) in the formulation may be 0.2%-51%, 0.2%-50%, 0.5-50%, 1%-40%, 5%-50%, 5%-40%, 5%-30%, 5%-25%, 5%-20%, 1%-30%, 1%-25%, 1%-20%, 1-10%, 0.5%-10%, 1%-5%, or ≤2%, such as 1%, 0.75%, or 0.5%.

[0138] In some embodiments, the acidic pH adjuster may include or be selected from hydrochloric acid, sulfuric acid, acetic acid, citric acid, phosphoric acid, malic acid, tartaric acid, lactic acid, and carbonic acid.

[0139] In some embodiments, in the above method, R is contained in a powder.

[0140] In some embodiments, in the above method, the powder consists of a dry powder containing R and a solvent.

[0141] In some embodiments, in the above method, the dry powder is R dry powder, and the solvent is water for injection.

[0142] In some embodiments, the formulation described above does not contain an acid that would cause N,N-dimethylbiguanide(R) in the aqueous solution to be significantly converted into metformin salt.

[0143] In the above method, the applied drug combination or composition comprises N,N-dimethylbiguanide (R) and component (Z), wherein the application concentration of N,N-dimethylbiguanide is 0.1%-51% by weight / w / w, preferably 0.2%-50%, and the drug includes a cell-responsive antitumor drug (abbreviated as B).

[0144] In some embodiments, in the above method, R in the drug combination is defined as being used to provide activities (abbreviated as activity A) that are difficult to provide by standard metformin (the active ingredient being a metformin salt, abbreviated as R'), including the effect of effectively damaging diseased tissue (abbreviated as activity A2); 1) Application to the lesion; 2) The dosage for one course of treatment is: In the formula, nR represents the number of times R is applied to the lesion in one course of treatment, qR(i) represents the dosage of R in the i-th application (i=1, ..., nR), and: In the formula, nRi is the number of application points of R at the i-th application, such as the number of injection points, cR(ii) and vR(ii) are the concentration and volume of R applied at the ii-th application point, such as the injection point, respectively, wherein the concentration threshold of cR(ii) (W / V) is 0.1% or 0.2%, and the upper limit of the concentration threshold is 51%, and the volume threshold of vR(ii) is 1 / 800 or 1 / 300 of the target volume (abbreviated as vtarget(ii)), and the upper limit of the volume threshold is 3 times vtarget(ii); - The B provides the cellular response (abbreviated as active B) by means of the following, one of the key control steps of which is crossing the aforementioned biological barrier: 1) Intratumoral application and / or systemic application; 2) The dosage for one course of treatment is: In the formula, QB is the standard dosage of B in tumors sensitive to B, nB and qB(j) are the number of systemic administrations of B in one course of treatment and the dosage of the j-th administration (j=1, ..., nB), respectively, and nB', cB(j'), and vB(j') are the number of tumor administrations of B in one course of treatment and the concentration and volume of the j'-th administration (j'=1, ..., nB'), respectively. - The type of combination is a synergistic combination in which the drug effects of activity A of R and activity B of B mutually enhance each other.

[0145] In some embodiments, the N,N-dimethylbiguanide (R) comprises <30%, preferably ≤5% metformin salt (R'), or does not include metformin salt (R'). Preferably, the N,N-dimethylbiguanide (R) is in the form of a powder for injection.

[0146] In some embodiments, in the above-described method, the component (Z) in the drug combination or composition includes an antitumor drug selected from cell-reactive drugs that cross the tumor tissue barrier as one of their key control steps.

[0147] In some embodiments, the cell-responsive drug includes cytotoxic drugs and targeted antitumor drugs.

[0148] In some embodiments, the cytotoxic drug includes DNA damaging agents, antimetabolites, microtubule inhibitors, and topoisomerase inhibitors.

[0149] In some embodiments, representative compounds of the DNA damaging agent include cisplatin, carboplatin, oxaliplatin, ifosfamide, and doxorubicin.

[0150] In some embodiments, representative compounds of the antimetabolite include fluorouracil, gemcitabine, and methotrexate.

[0151] In some embodiments, representative compounds of the microtubule inhibitor include paclitaxel and vincristine.

[0152] In some embodiments, the targeted antitumor drug in the above method includes a kinase inhibitor.

[0153] In some embodiments, the kinase inhibitors in the above methods include gefitinib, osimertinib, larotrectinib, pazopanib, trastuzumab, and their derivatives.

[0154] In some embodiments, in the above method, B comprises or is selected from at least one of the following: cisplatin, cyclophosphamide, carboplatin, doxorubicin, ifosfamide, nitrogen mustard, fluorouracil, capecitabine, gemcitabine, methotrexate, pemetrexed, fludarabine, paclitaxel, docetaxel, vincristine, vinca, vindesin, etoposide, irinotecan, topotecan, daunorubicin, mitoxantrone, gefitinib, osimertinib, larotrectinib, pazopanib, trastuzumab and its derivatives.

[0155] In some embodiments, the application concentration (w / w) of the N,N-dimethylbiguanide (R) in the above method can be 0.2%-51%, 0.2%-50%, 0.5-50%, 0.5-45%, 1%-45%, 5%-45%, 5%-40%, 5%-30%, 5%-25%, 5%-20%, 1%-30%, 1%-25%, 1%-20%, 0.5%-10%, 1%-5%, or ≤2%, such as 1%, 0.75%, or 0.5%.

[0156] In some embodiments, in the above method, the drug combination or composition includes dimethyl biguanide (R) and doxorubicin (DOX), wherein the dimethyl biguanide (R) can be administered at a concentration of 0.2%-50% (w / w), and the doxorubicin (DOX) can be administered at a concentration of 0.025%-1% (w / w). Preferably, the dimethyl biguanide (R) is administered at a concentration of 5%-50% (w / w), and the doxorubicin (DOX) is administered at a concentration of 0.1%-1%. Optionally, both dimethyl biguanide (R) and doxorubicin (DOX) are administered to the tumor.

[0157] In one embodiment, the dimethyl biguanide (R) is administered to the tumor at a concentration of 0.2%-50% (w / w), the doxorubicin (DOX) is administered to the tumor at a concentration of 0.025%-1% (w / w), and the doxorubicin (DOX) is administered systemically at a concentration of 0.05%-0.06% (w / w), such as intraperitoneally.

[0158] In some embodiments, in the above method, the drug combination or composition includes dimethylbiguanide(R) and ifosfamide (IFO), wherein the dimethylbiguanide(R) can be applied at a concentration of 0.2%-50% (w / w), and the ifosfamide (IFO) can be applied at a concentration of 0.1%-10% (w / w). Preferably, the dimethylbiguanide(R) is applied at a concentration of 0.2%-45% (w / w), and the ifosfamide (IFO) is applied at a concentration of 0.1%-10% (w / w). More preferably, the dimethylbiguanide(R) is applied at a concentration of 5%-45% (w / w), and the ifosfamide (IFO) is applied at a concentration of 0.5%-10% (w / w). Optionally, both dimethylbiguanide(R) and ifosfamide (IFO) are applied to the tumor.

[0159] In one embodiment, the dimethyl biguanide (R) is administered to the tumor at a concentration of 5%-50% (w / w), the ifosfamide (IFO) is administered to the tumor at a concentration of 0.5%-10% (w / w), and the ifosfamide (IFO) is administered systemically at a concentration of 0.5% (w / w), such as intraperitoneally.

[0160] In some embodiments, in the above method, the drug combination or composition includes dimethyl biguanide (R) and cisplatin (DDP), wherein the dimethyl biguanide (R) can be administered at a concentration (w / w) of 0.2%-50%, and the cisplatin (DDP) can be administered at a concentration (w / w) of 0.01%-0.4%. Preferably, the dimethyl biguanide (R) is administered at a concentration (w / w) of 0.5%-50%, and the cisplatin (DDP) is administered at a concentration (w / w) of 0.01%-0.04%. Optionally, both dimethyl biguanide (R) and cisplatin (DDP) are administered to the tumor.

[0161] In one embodiment, the dimethyl biguanide (R) is administered to the tumor at a concentration of 0.5%-50% (w / w), and the cisplatin (DDP) is administered to the tumor at a concentration of 0.013%-0.4% (w / w). Preferably, the cisplatin (DDP) is administered to the tumor system at a concentration of 0.015% (w / w), such as intraperitoneally.

[0162] In some embodiments, in the above method, the drug combination or composition includes dimethylbiguanide (R) and 5-fluorouracil (5-FU), wherein the dimethylbiguanide (R) can be applied at a concentration (w / w) of 0.2%-50%, and the 5-fluorouracil (5-FU) can be applied at a concentration (w / w) of 0.05%-9%. Preferably, the dimethylbiguanide (R) is applied at a concentration (w / w) of 0.2%-45%, and the 5-fluorouracil (5-FU) is applied at a concentration (w / w) of 0.5%-9%. More preferably, the dimethylbiguanide (R) is applied at a concentration (w / w) of 5%-45%, and the 5-fluorouracil (5-FU) is applied at a concentration (w / w) of 0.9%-9%. Optionally, both dimethylbiguanide (R) and 5-fluorouracil (5-FU) are applied to the tumor.

[0163] In one embodiment, the dimethyl biguanide (R) is administered to the tumor at a concentration of 5%-45% (w / w), the 5-fluorouracil (5-FU) is administered to the tumor at a concentration of 0.9%-9% (w / w), and the 5-fluorouracil (5-FU) is administered systemically at a concentration of 0.5% (w / w), such as intraperitoneally.

[0164] In some embodiments, in the above methods, the drug combination or composition includes dimethylbiguanide (R), cisplatin (DDP), and gemcitabine (GEM), wherein the dimethylbiguanide (R) can be administered at a concentration of 0.2%-50% (w / w), the cisplatin (DDP) can be administered at a concentration of 0.05%-1% (w / w), and the gemcitabine (GEM) can be administered at a concentration of 0.5-2% (w / w), preferably at a concentration of 1% (w / w). Optionally, or dimethylbiguanide (R) and cisplatin (DDP) can be administered to the tumor and gemcitabine (GEM) can be administered systemically, such as intraperitoneally.

[0165] In some embodiments, in the above methods, the drug combination or composition includes dimethyl biguanide (R), paclitaxel (PTX), and 5-fluorouracil (5-FU), wherein the dimethyl biguanide (R) can be administered at a concentration of 0.2%-50% (w / w), the 5-fluorouracil (5-FU) can be administered at a concentration of 0.1%-10% (w / w), and the paclitaxel can be administered at a concentration of 0.02-1% (w / w), preferably at a concentration of 0.03% (w / w). Optionally, the dimethyl biguanide (R) and 5-fluorouracil (5-FU) are administered intratumorally and the paclitaxel (PTX) is administered systemically, such as intraperitoneally.

[0166] In some embodiments, in the above methods, the drug combination or composition includes bimethyl biguanide (R) and osimertinib, wherein the bimethyl biguanide (R) can be administered at a concentration (w / w) of 0.2%-50%, and the osimertinib can be administered at a concentration (w / w) of 0.05%-2%. Preferably, the bimethyl biguanide (R) is administered at a concentration (w / w) of 5%, and the osimertinib is administered at a concentration (w / w) of 0.15%. Optionally, the bimethyl biguanide (R) is administered via vomiting, and the osimertinib is administered systemically, such as via intraperitoneal administration.

[0167] In some embodiments, in the above methods, the drug combination or composition includes dimethylbiguanide (R) and gefitinib (GR), wherein the dimethylbiguanide (R) can be administered at a concentration (w / w) of 0.2%-50%, and the gefitinib (GR) can be administered at a concentration (w / w) of 0.1%-2%. Preferably, the dimethylbiguanide (R) is administered at a concentration (w / w) of 1%-50%, and the gefitinib (GR) is administered at a concentration (w / w) of 0.5%. Optionally, the dimethylbiguanide (R) is administered to the tumor, and the gefitinib (GR) is administered systemically, for example, intraperitoneally.

[0168] In some embodiments, in the above method, the drug combination includes dimethyl biguanide (R), cisplatin (DDP), and gefitinib (GR), wherein the dimethyl biguanide (R) is administered at a concentration of 5% (w / w) (e.g., in the tumor), the gefitinib (GR) is administered at a concentration of 0.5% (w / w) (systemically, e.g., intraperitoneally), and the cisplatin (DDP) is administered at a concentration of 0.15% (w / w) (e.g., in the tumor).

[0169] In some embodiments, in the above methods, the drug combination or composition includes dimethyl biguanide (R), ifosfamide (IFO), and gefitinib (GR), wherein the dimethyl biguanide (R) is administered at a concentration of 5% (w / w) (e.g., in the tumor), the ifosfamide (IFO) is administered at a concentration of 1% (w / w) (e.g., in the tumor), and the gefitinib (GR) is administered at a concentration of 0.5% (w / w) (systemically, e.g., intraperitoneally).

[0170] In some embodiments, in the above methods, the drug combination package or composition includes dimethyl biguanide (R), doxorubicin (DOX), and gefitinib (GR), wherein the dimethyl biguanide (R) is administered at a concentration of 5% (w / w) (e.g., in the tumor), the doxorubicin (DOX) is administered at a concentration of 0.05% (w / w) (e.g., in the tumor), and the gefitinib (GR) is administered at a concentration of 0.5% (w / w) (systemically, e.g., intraperitoneally).

[0171] In some embodiments, in the above method, the drug combination or composition includes dimethyl biguanide (R) and sodium bicarbonate (SB), wherein dimethyl biguanide (R) can be applied at a concentration of 0.5%-4%, and sodium bicarbonate (SB) can be applied at a concentration of 0.5%-15%, preferably 0.5%-10%, more preferably 1.5%-9%; preferably dimethyl biguanide (R) is applied at a concentration of 1%-3%, and sodium bicarbonate (SB) is applied at a concentration of 1%-9%; more preferably, dimethyl biguanide (R) is applied at a concentration of 3%, and sodium bicarbonate (SB) is applied at a concentration of 3%-9%. Optionally, the drug combination further includes ifosfamide (IFO) at a concentration of 0.5%, 5-fluorouracil (5-FU) at a concentration of 0.4%, or osimertinib at a concentration of 0.3%. Preferably, the above-mentioned drug combination containing dimethyl biguanide (R) and sodium bicarbonate (SB) is administered to the tumor.

[0172] In some embodiments, the pharmaceutical combination or composition described above includes dimethylbiguanide (R) and a methylene blue dye (D) such as methylene blue (MB). In one embodiment, the pharmaceutical combination includes dimethylbiguanide (R) and methylene blue (MB), wherein the dimethylbiguanide (R) can be applied at a concentration of 1%-50%, and the MB can be applied at a concentration of 0.3%-5%, preferably 0.5%-5%; more preferably, the dimethylbiguanide (R) is applied at a concentration of 5%-50%, and the MB is applied at a concentration of 0.5%-5%; preferably, the dimethylbiguanide (R) is applied at a concentration of 5%-50%, and the MB is applied at a concentration of 0.5%-1%. Optionally, the pharmaceutical combination further includes one or more of ifosfamide (IFO) at a concentration of 0.5%-1%, 5-fluorouracil (5-FU) at a concentration of 0.4%-1%, and osimertinib at a concentration of 0.3%. Preferably, the drug combination comprising bimethyl biguanide (R) and methylene blue (MB) can be administered intratumorally and / or systemically. In some embodiments, the drug combination comprises bimethyl biguanide (R), MB, and any one of IFO, 5-FU, and osimertinib, wherein bimethyl biguanide (R) is administered at a concentration of 5%, MB at a concentration of 1%, IFO at a concentration of 0.5%, 5-FU at a concentration of 0.4%, and osimertinib at a concentration of 0.3%. In some embodiments, in the above method, the drug combination comprises bimethyl biguanide (R), SB, MB, and any one of IFO, 5-FU, and osimertinib, wherein bimethyl biguanide (R) is administered at a concentration of 5%, SB at a concentration of 5%-10%, MB at a concentration of 0.5%, IFO at a concentration of 0.5%, 5-FU at a concentration of 0.4%, and osimertinib at a concentration of 0.3%.

[0173] In some embodiments, the drug combination in the above method includes dimethyl biguanide (R) and an immunomodulator (E), said immunomodulator including or selected from macromolecular immunomodulators (E1), cellular immunomodulators (E2), and vaccine-type immunomodulators (E3). In some embodiments, the macromolecular immunomodulator can be, for example, an immune checkpoint inhibitor, a cytokine, and a TLR agonist. In one embodiment, the immune checkpoint inhibitor can be Anti-PD-1 (CD279), the cytokine can be interleukin-15, and the TLR agonist can be CpG ODN 1826 (CpG), and the vaccine-type immunomodulator can be BCG.

[0174] In some embodiments, in the above method, the drug combination includes dimethylbiguanide(R) and CpG ODN 1826 (CpG), wherein dimethylbiguanide(R) can be administered at a concentration of 1%-50%, and CpG can be administered at a concentration of 0.1%; preferably, dimethylbiguanide(R) is administered at a concentration of 2.5%-50%, and CpG is administered at a concentration of 0.1%. Preferably, the drug combination or composition further includes MB at a concentration of 1%, IFO at a concentration of 0.5%, and / or SB at a concentration of 10%. In some embodiments, the drug combination or composition comprises dimethylbiguanide(R) and CpG and any one or two of the following: IFO, MB, and SB, wherein dimethylbiguanide(R) is administered at a concentration of 2.5%-50%, IFO at a concentration of 0.5%, CpG at a concentration of 0.1%, MB at a concentration of 1%, and SB at a concentration of 10%.

[0175] In some embodiments, in the above methods, the drug combination or composition includes dimethyl biguanide (R) and an immune checkpoint inhibitor Anti-PD-1 (CD279), such as RMP1-14, wherein dimethyl biguanide (R) may be administered at a concentration of 0.2%-50%, and RMP1-14 may be administered at a concentration of 0.1%; preferably, dimethyl biguanide (R) is administered at a concentration of 1%-50%, and RMP1-14 is administered at a concentration of 0.1%.

[0176] In some embodiments, in the above method, the drug combination or composition includes dimethylbiguanide(R) and BCG, wherein dimethylbiguanide(R) can be administered at a concentration of 1%-50%, preferably 2.5%-50%, more preferably 5%-50%, and BCG can be administered at an amount of 1×10⁶ CFU; preferably, dimethylbiguanide(R) is administered at a concentration of 50%, and BCG is administered at an amount of 1×10⁶ CFU. Optionally, the drug combination or composition further includes either ifosfamide (IFO) at a concentration of 0.5% or SB at a concentration of 10%. In some embodiments, the drug combination or composition comprises dimethylbiguanide(R) and BCG and either IFO or SB, wherein dimethylbiguanide(R) is administered at a concentration of 5%-50%, BCG is administered at an amount of 1×10⁶ CFU, IFO is administered at a concentration of 0.5%, and SB is administered at a concentration of 10%.

[0177] In some embodiments, in the above methods, the drug combination or composition includes dimethyl biguanide (R) and an immunomodulator (E), said immunomodulator including or selected from BCG, CpG ODN 1826 (CpG), and immune checkpoint inhibitors such as anti-PD-1.

[0178] In some embodiments, in the above method, the drug combination or composition includes dimethyl biguanide (R) and BCG, wherein dimethyl biguanide (R) can be administered at a concentration of 1%-50%, preferably 2.5%-50%, more preferably 5%-50%, and BCG can be administered at an amount of 1×10⁶ CFU; preferably, dimethyl biguanide (R) is administered at a concentration of 50%, and BCG is administered at an amount of 1×10⁶ CFU. Optionally, the drug combination or composition further includes any one of ifosfamide (IFO) at a concentration of 0.5%, MB at a concentration of 1%, and SB at a concentration of 10%. In some embodiments, the drug combination or composition comprises dimethylbiguanide (R) and BCG and any one of the following: IFO, MB, and SB, wherein dimethylbiguanide (R) is administered at a concentration of 50%, BCG is administered at an amount of 1 × 10⁶ CFU, MB is administered at a concentration of 1%, IFO is administered at a concentration of 0.5%, and SB is administered at a concentration of 10%.

[0179] In some embodiments, in the above methods, the drug combination or composition includes dimethylbiguanide(R) and CpG ODN 1826 (CpG), wherein dimethylbiguanide(R) can be administered at a concentration of 1%-50%, preferably 2.5%-50%, and CpG can be administered at a concentration of 0.1%; preferably, dimethylbiguanide(R) is administered at a concentration of 2.5%-50%, and CpG is administered at a concentration of 0.1%. Preferably, the drug combination or composition further includes MB at a concentration of 1% or SB at a concentration of 10%. In some embodiments, the drug combination or composition comprises dimethylbiguanide(R) and CpG and any one of the following: MB and SB, wherein dimethylbiguanide(R) is administered at a concentration of 50%, CpG at a concentration of 0.1%, MB at a concentration of 1%, and SB at a concentration of 10%.

[0180] Preferably, in the above method, the drug combination or composition containing dimethyl biguanide (R) and immunomodulators such as BCG, anti-PD-1 or CpG ODN 1826 (CpG) is administered intralesionally or to the tumor, or intralesionally or to the tumor and systemically, such as by intraperitoneal injection or intravenous injection or administration to the lesion or tumor.

[0181] In some preferred embodiments, the contact, application, or intervention in the above method may include micro-volume multi-point contact, application, or intervention. Preferably, it is micro-volume multi-point injection, wherein the micro-volume can be a drug application volume of ≤5% or 10% of the target volume, for example, including amounts of 5μL-1000μL, such as 5μL-500μL, 5μL-250μL, 5μL-200μL, 5μL-100μL, 5μL-100μL, etc. The dosage range is 75μL, 5μL-50μL, 5μL-25μL, 5μL-20μL, 10μL-200μL, 10μL-100μL, 10μL-75μL, 20μL-200μL, 20μL-100μL, 20μL-75μL, 30μL-200μL, 30μL-100μL, 40μL-200μL, 40μL-100μL, 50μL-200μL, 50μL-100μL, or 50μL-75μL, etc. Preferably, it is applied through micro-volume multi-point contact or administration; more preferably, it is injected through micro-volume multi-point injection.

[0182] In some implementations, the above-described treatment, intervention, or therapy may be sustained for at least one course of treatment. A course of treatment may last for approximately 7-40 days, for example, approximately 9 to approximately 37 days, such as approximately 15, 20, 25, 27, 28, 29, 30, 31, 32, 33, 34, or 35 days.

[0183] In one embodiment, during a course of treatment, contact with or application of the formulation of the present invention may be at a low frequency, such as about 1-6 times per course of treatment, preferably 1-5 times per course of treatment, 1-4 times per course of treatment, 1-3 times per course of treatment, or 1-2 times per course of treatment.

[0184] In some embodiments, in the above method, the dimethyl biguanide (R) and component (Z) in the drug combination can be administered simultaneously, sequentially, or at intervals.

[0185] In some embodiments, the drug combination, formulation, or composition can be applied topically in the above methods.

[0186] In some embodiments, in the above method, both dimethyl biguanide (R) and component (Z) in the drug combination are administered intralesionally or intratumorally (combination relationship I); R and Z are administered intralesionally or intratumorally and systemically, respectively (combination relationship II); or R and part of Z are administered intralesionally or intratumorally, and another part of Z is administered systemically (combination relationship III).

[0187] In some embodiments, in the above method, the tumor is a solid tumor suitable for application to the tumor body, preferably applied locally.

[0188] In some embodiments, in the above method, the tumor is selected from tumors whose tissue barrier function is unfavorable to the cell action of Z, preferably from refractory tumors of Z, including chemotherapy-resistant tumors, tumors with unfavorable microenvironments, tumors that have been discontinued from antitumor drugs, tumors for which there are no effective chemotherapy drugs, and tumors carried by patients with contraindications or incompatibilities to standard chemotherapy drugs.

[0189] In some embodiments, in the above method, the tumor is selected from heterogeneous tumors, wherein the heterogeneity includes at least one of tissue heterogeneity, spatial heterogeneity, and safety heterogeneity. The heterogeneous tumor is a tumor in which different tumors of the same type or different regions of the same tumor are set as different target areas for differentiated treatment based on the heterogeneity.

[0190] In some embodiments, the contact or application in the above method includes multi-point contact or application, preferably micro-volume multi-point contact or application, and more preferably micro-volume multi-point injection.

[0191] In some embodiments, the trace amount includes a microvolume.

[0192] In some embodiments, in the above methods, the formulation or drug combination is suitable for application to microvolume administration or intervention to reduce (Y) lesion volume (X), wherein the ratio of the administration volume to the target volume (V_administered / V_target) is approximately 0.02-0.056, 0.02-0.10, or 0.02-0.34.

[0193] In some embodiments, in the above method, when the lesion is ≥3.5cm³ or the maximum size is ≥2cm, the number of points is ≥5 points, such as 10, 15, 20, 25, 30, 35 or 50 points.

[0194] In some embodiments, the contact or application in the above method is low-frequency, preferably 1-6 times per treatment course, more preferably 1-5 times per treatment course, 1-4 times per treatment course, 1-3 times per treatment course, or 1-2 times per treatment course.

[0195] In some embodiments, in the above method, preferably, the application concentration CR of R is ≥5%, ≥10%, ≥20%, or ≤30%; and the number of application points is >3, 10, 15, 20, 25, 30, 35, or 100.

[0196] In some embodiments, in the above method, the metformin R and component Z may be provided by one or more formulations, and wherein: R and Z are applied within the lesion or tumor; R and Z refer to intralesional or systemic administration, respectively; or The R and part of the Z are administered intralesionally, and another part is administered via the Z system.

[0197] In some embodiments, in the above method, R and Z work together, and wherein - The synergy refers to the ability of the following effects to be provided relative to the standard combined regimen (R' / Z combination) when the metformin dosage is at least halved (QR ≦ 1 / 2QR') and the Z dosage is at least 20% reduced: 1) Improved drug performance, including adaptation to tumor heterogeneity, manifested as an improvement of at least one drug adjustability index by at least 100%, wherein drug adjustability refers to the ability of a drug to provide differentiated administration parameters to different target areas while maintaining efficacy; and / or 2) Enhanced drug efficacy, manifested by an increase of at least 40% in at least one drug efficacy indicator.

[0198] In some embodiments, the synergy in the above method further includes: at least one of the drug adjustability indicators is increased by at least 10% and / or at least one of the drug efficacy indicators is enhanced by at least 10% relative to the R and B.

[0199] In some embodiments, in the above uses and methods, the N,N-dimethylbiguanide (R) is defined by the following to provide an active form that distinguishes it from standard metformin (whose active ingredient is R'): 1) Do not mix with acids that cause R to be significantly converted to R'; 2) Application to local lesions; 3) The dosage for one course of treatment is: In the formula, QR < 50% or 25% of the equivalent dosage of R', nR is the number of times R is applied to the lesion or tumor in one course of treatment, qR(i) is the dosage of R in the i-th application (i = 1, ..., nR), and: In the formula, nRi is the number of application points of R at the i-th application, such as the number of injection points, cR(ii) and vR(ii) are the concentration and volume of R applied at the ii-th application point, such as the injection point, respectively, wherein the concentration threshold of cR(ii) (W / V) is 0.1% or 0.2%, and the upper limit of the concentration threshold is 51%; the volume threshold of vR(ii) is 1 / 800 or 1 / 300 of the target volume (abbreviated as vtarget(ii)), and the upper limit of the volume threshold is 3 times vtarget(ii).

[0200] The active form of R' is characterized by complete complexation with acid in solution; while the active form of R is characterized by less complexation with acid in solution, mainly existing in a form of very little or partial complexation. Thus, through the transient local action of this different active form, it provides the activity that R' cannot provide (abbreviated as activity A), including at least one of the following: the effect of effectively reducing the function of biological barriers (abbreviated as activity A1), the effect of effectively damaging diseased tissue (abbreviated as activity A2), and / or the effect of effectively releasing diseased immune substances and / or inflammatory signals within the lesion (abbreviated as activity A3).

[0201] In some embodiments, in the above uses and methods, R and drug (Z) are used together, wherein the application concentration (w / w) of R is 0.1%-51%, preferably 0.2%-50%, and the drug (Z) comprises at least one selected from the group consisting of: cell-responsive antitumor drugs (abbreviated as B), near-neutral or weakly basic sodium salts (abbreviated as C), methylene blue (abbreviated as D), and immunomodulators (abbreviated as E), wherein: - The cell-responsive antitumor drug (B) provides the cell response (abbreviated as active B) by means of the following, one of the key controlled steps of which is crossing the aforementioned biological barrier: 1) Intralesional application and / or systemic application; 2) The dosage for one course of treatment is: , In the formula, QB is the standard dosage ≤ in the sensitive tumors of B, nB and qB(j) are the number of systemic administrations of B in one course of treatment and the dosage of the j-th administration (j=1, ..., nB), respectively, and nB', cB(i'), and vB(i') are the number of tumor administrations of B in one course of treatment and the concentration and volume at the j'-th administration (j'=1, ..., nB'), respectively; - The C is used to provide an effect that improves the local efficacy-toxicity ratio of the R (abbreviated as active C) by the following definition: 1) Apply in combination with the R; 2) The mixing ratio of C to R (QC / QR) is 0.5 to 4.0; - The D provides pharmacological efficacy that enhances the drug effect of the R by the following limitations: 1) Application to lesions, including application in combination with or separately from the R; 2) The dosage QE for one course of treatment is ≤ the effective dosage for transient local effects. The C is used to provide an effect that improves the local efficacy-toxicity ratio of the R (abbreviated as Activity C) by the following limitations: 1) Apply in combination with the R; 2) The mixing ratio of C to R (QC / QR) is 0.5 to 4.0; - The D provides pharmacological efficacy that enhances the drug effect of the R by the following limitations: 1) Application to lesions, including application in combination with or separately from the R; 2) The dosage (QE) for one course of treatment is ≤ the effective dosage for transient local effects. - The E is defined by the following functions for providing immunomodulatory effects benefiting from the effects of the active A1, and / or optimizing local and / or systemic immune responses by utilizing the effects of the active A3 (abbreviated as active E): 1) Intralesional application and / or systemic application; 2) The dosage for one course of treatment is: In the formula, QE is the standard dosage of E in the anti-lesion application, nE and qE(k) are the number of systemic applications of E in one course of treatment and the dosage of the kth application (j=1, ...nk), respectively, and nE', cE(k') and vE(k') are the number of tumor applications of E in one course of treatment and the concentration and volume at the k'th application (k'=1, ...nk'), respectively.

[0202] In some embodiments, in the above uses and methods, R is used in conjunction with cell-responsive antitumor drugs (abbreviated as B) and immunomodulators (abbreviated as E), wherein: - The cell-responsive antitumor drug (B) provides the cell response (abbreviated as active B) by means of the following, one of the key controlled steps of which is crossing the aforementioned biological barrier: 1) Intralesional application and / or systemic application; 2) The dosage for one course of treatment is: , In the formula, QB is the standard dosage ≤ of B in sensitive tumors, nB and qB(j) are the number of systemic administrations of B in one course of treatment and the dosage of the j-th administration (j=1, ..., nB), respectively, and nB', cB(i'), and vB(i') are the number of tumor administrations of B in one course of treatment and the concentration and volume at the j'-th administration (j'=1, ..., nB'), respectively. - The immunomodulator (E) is defined by the following: to provide immunomodulatory effects benefiting from the effects of said active A1, and / or to optimize local and / or systemic immune responses by utilizing the effects of said active A3 (abbreviated as active E): 1) Intralesional application and / or systemic application; 2) The dosage for one course of treatment is: In the formula, QE is the standard dosage of E in the anti-lesion application, nE and qE(k) are the number of systemic applications of E in one course of treatment and the dosage of the kth application (j=1, ...nk), respectively, and nE', cE(k') and vE(k') are the number of tumor applications of E in one course of treatment and the concentration and volume at the k'th application (k'=1, ...nk'), respectively.

[0203] In some embodiments, the drug adjustability index in the above method includes at least one of the following tumor administration adjustability coefficients: 1) The concentration coefficient = the ratio of the highest to the lowest value of cR(ii), and the coefficient is >50, 100 or 150, which allows the combination to provide highly variable application concentrations of R to different tumors of the same type and / or different regions of the same tumor while maintaining its said activity A stable; 2) Volume ratio coefficient = the ratio of the highest to the lowest volume ratio of vR(ii) to vtarget(ii), and said coefficient is >5, 10, or 100, which allows the combination to provide highly variable application volume ratios of R to different tumors of the same type and / or different regions of the same tumor while maintaining its said activity A stable; and 3) Application point density coefficient = the ratio of the highest to the lowest value of nRi within the target (ii), and the coefficient is >5, 10 or 100, which enables the combination to provide highly variable needle point density R to different tumors of the same type or / and different regions of the same tumor while maintaining its active A stable.

[0204] In some embodiments, the drug efficacy indicator in the above method includes at least one of the following: 1) Timeliness indicator, expressed as the time to significant effect in mouse trials (the day when the tumor proliferation rate is <42%), and the time to significant effect is reduced by at least 25% compared to the R' regimen, the R' / Z combined regimen, and the Z monotherapy regimen, which makes the combination a rapid-acting or even more rapid-acting combination, especially suitable for tumors that require rapid reduction in tumor size; 2) Efficacy indicators, expressed as tumor inhibition rate or objective response rate in mouse trials, wherein the tumor inhibition rate or objective response rate is at least 25% higher than that of the R' regimen, the R' / Z co-treatment regimen, and the Z monotherapy regimen, making the combination a highly effective or even more highly effective combination, especially suitable for tumors that require maximum inhibition of tumor size; 3) Long-acting indicator, manifested as progression-free time in mouse trials, wherein the progression-free time is extended by at least 25% compared to the R' regimen, the R' / Z co-treatment regimen, and the Z monotherapy regimen, which makes the combination a long-acting or further long-acting combination, especially suitable for tumors in which the progression-free time needs to be extended as much as possible.

[0205] According to a twelfth aspect of the present invention, a kit is provided comprising the above-described formulations, pharmaceutical combinations or compositions of the present invention, and instructions for carrying out the above-described methods of the present invention.

[0206] In some embodiments, the kit further includes a microvolume intervention device.

[0207] In some embodiments, the microvolume intervention device in the above-described kit is suitable for micro-volume multi-point contact or application, and preferably includes a micro-injection device.

[0208] According to a thirteenth aspect of the present invention, a method for preparing a formulation is provided, comprising: selecting N,N-dimethylbiguanide with a metformin salt content of <30%, ≤25%, preferably ≤10%, more preferably ≤5% as a raw material; excluding and avoiding contact in the formulation with an acidic pH adjuster that would enable the N,N-dimethylbiguanide to meet the requirements of cell culture or standard injection safety; using a strong alkali resistant preparation system; and dispersing the N,N-dimethylbiguanide in a solvent.

[0209] In some embodiments, the solvent is water.

[0210] In some embodiments, the N,N-dimethylbiguanide is dissolved in water.

[0211] According to the fourteenth aspect of the invention, the use of N,N-dimethylbiguanide as a transient active ingredient in the preparation of formulations, pharmaceuticals, pharmaceutical combinations or kits for treating tumors or nodules is provided, wherein the N,N-dimethylbiguanide is used to provide a transient effect on diseased tissue and its contained cells.

[0212] In this application, the terms "transient activity" or "transient effect" refer to the activity or effect provided during a transient period before the active form of R is converted to the active form of R' in vivo.

[0213] In one embodiment, N,N-dimethylbiguanidine is an organic compound with the chemical formula C4H11N5 having Formula 1 as described in the first aspect above.

[0214] According to a fifteenth aspect of the present invention, a method for treating lesion cells in a subject is provided, comprising administering the above-described N,N-dimethylbiguanide of the present invention or an agent of the present invention into the lesion, penetrating the lesion, and contacting the lesion tissue cells, wherein the N,N-dimethylbiguanide serves as a transient active ingredient to provide a transient effect on the lesion tissue and its contained cells, the transient effect being, for example, a transient tissue penetration effect.

[0215] According to a sixteenth aspect of the present invention, a method for treating lesions in a subject is provided, comprising: applying the N,N-dimethylbiguanide or an agent of the present invention described above to the lesion, penetrating into the lesion tissue, and contacting with components of the lesion tissue, or applying the agent of the present invention to the lesion tissue. Preferably, the application or contact can be systemic or local, such as by injection into a tumor or tumor microenvironment.

[0216] According to a seventeenth aspect of the invention, a method for treating lesions such as tumors or nodules in a subject is provided, comprising intervening in or administering to the subject the pharmaceutical combination, composition, or formulation described in the invention as above.

[0217] In some embodiments, the intervention or application can be systemic or local. Preferably, the application or intervention can include intratumoral application or intervention, preferably local intratumoral intervention, preferably local injection application or intervention into the tumor or tumor microenvironment.

[0218] In some implementations, injection administration may include infusion, liquid particle injection, or implantation.

[0219] In some preferred embodiments, the formulations described above are suitable for micro-volume administration or interventional administration, wherein the ratio of the administration volume to the target volume (V_administered / v_target) is approximately 0.02-0.05, 0.02-0.10, or 0.02-0.34.

[0220] In some embodiments, the above-described contact, intervention, or application includes multi-point contact, intervention, or application, preferably local multi-point contact or intervention.

[0221] The aforementioned contact or application may include multiple (site) contact or application or intervention. The multiple sites refer to the number of micro-volume intervention sites where the total target area efficacy evaluation is observed to be effective, which is significantly higher than that of the standard metformin drug. Specifically, the multiple sites are, for example, multiple sites at the target area, such as ≥5, ≥10, ≥20, ≥30, ≥50 sites, such as at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15 or more sites, such as 10 to 20, 10-30, 10-40, 10-50, 20-30, 20-40, 20-50, 25-50, 30-50 or more sites. Preferably, the contact points or application sites or intervention points are evenly distributed, especially to areas where standard intervention cannot diffuse.

[0222] In some preferred embodiments, when the lesion is ≥3.5 cm3 or the maximum size is ≥2 cm, the number of sites is ≥5, such as 10, 15, 20, 25, 30, 35 or 50 sites.

[0223] In some preferred embodiments, the aforementioned contact, application, or intervention may include micro-volume multi-point contact, application, or intervention, preferably micro-volume multi-point injection, wherein the micro-volume is a drug application volume ≤5% or 10% of the target volume, for example, including amounts from 5 μL to 1000 μL, such as 5 μL-500 μL, 5 μL-250 μL, 5 μL-200 μL, 5 μL-100 μL, 5 μL-75 μL, 5 μL-50 μL, 5 μL -25μL, 5μL-20μL, 10μL-200μL, 10μL-100μL, 10μL-75μL, 20μL-200μL, 20μL-100μL, 20μL-75μL, 30μL-200μL, 30μL-100μL, 40μL-200μL, 40μL-100μL, 50μL-200μL, 50μL-100μL, or 50μL-75μL, etc. Preferably, it is applied by micro-volume multi-point contact or administration; more preferably, it is injected by micro-volume multi-point injection.

[0224] In some implementations, the above treatment can be sustained for at least one course of treatment. The duration of one course of treatment can be approximately 7 to 40 days, for example, approximately 9 to approximately 37 days, for example, approximately 15, 20, 25, 27, 28, 29, 30, 31, 32, 33, 34, or 35 days.

[0225] In one embodiment, during a course of treatment, contact with or application of the formulation of the present invention may be at a low frequency, such as about 1-6 times per course of treatment, preferably 1-5 times per course of treatment, 1-4 times per course of treatment, 1-3 times per course of treatment, or 1-2 times per course of treatment.

[0226] According to the eighteenth aspect of the present invention, a kit is provided comprising the formulation, pharmaceutical combination or composition as described above, and instructions for carrying out the treatment or treatment method as described above.

[0227] In one embodiment, the kit of the present invention further includes a micro-volume intervention device, preferably suitable for micro-volume multi-point contact or application, preferably including a micro-injection device such as a syringe.

[0228] In one embodiment, the volumetric intervention device may include, for example, a puncture needle, an injection needle, a catheter, or other devices with the same function.

[0229] In one embodiment, in the above-described uses, methods, and kits of the present invention, the dosage (QR) of R in the formulation of the present invention for one course of treatment is ≤20% or 50% of the dosage (QR') of standard metformin drug (R'), wherein R is a transient active ingredient that exceeds the expectation of "R and R' being pharmaceutically equivalent", characterized by a significant reduction in dosage but a significant increase in drug effect, including an increase of at least one drug adjustability indicator by at least 100% and an enhancement of at least one drug efficacy indicator by at least 40%.

[0230] In this application, the terms "transient activity" or "transient effect" refer to the local activity or local effect provided during a transient period before the active form of R is converted to the active form of R' in vivo.

[0231] In one embodiment, in the above-described uses, methods, and kits of the present invention, the transient dosage (QR and QZ) of the combination of R and Z (abbreviated as R / Z) in the formulation of the present invention is significantly less than the dosage (QR' and QZ') of the combination of standard metformin drug (R') and Z (abbreviated as R' / Z): QR ≤ 25% or 50% QR' and QZ not greater than QZ', wherein R / Z is a synergistic combination exceeding the expectation of "R / Z and R' / Z being a pharmaceutically equivalent combination", characterized by a significant reduction in dosage but a significant increase in drug effect, including an increase of at least one drug adjustability index by at least 100% and an enhancement of at least one drug efficacy index by at least 25%, wherein drug adjustability refers to the ability of a drug to provide differentiated dosing parameters to different target areas while maintaining efficacy.

[0232] In one embodiment, in the above-described uses, methods, and kits of the present invention, at least one of the amounts (QR and QB) of the combination of R and Z (abbreviated as R / Z) in the formulation of the present invention is lower than the amounts (QR0 and QZ0) of R and Z monotherapies: QR is not greater than QR0 and QZ ≤ 80% QB0, wherein the R / Z is a synergistic combination exceeding the expectations of the HSA or Bliss method based on the effects of R and Z monotherapies, characterized by a reduction in the amount but an increase in the drug effect, including an increase of at least 10% in at least one drug adjustability indicator and an enhancement of at least 10% in at least one drug efficacy indicator.

[0233] In one embodiment, in the above-described uses, methods, and kits of the present invention, the intervention or administration concentration (C, w / w) of N,N-dimethylbiguanide in the formulation of the present invention is 1.0% ≦C ≦30%, for example 1.0% ≦C ≦20%, preferably 3.0% ≦C ≦30%, more preferably 3.0% ≦C ≦20%, more preferably 5.0% ≦C ≦30%, more preferably 5.0% ≦C ≦20%, and particularly preferably 6.0% ≦C ≦20%.

[0234] In one embodiment, in the above-described uses, methods, and kits of the present invention, the intervention or administration concentration (C, w / w) of N,N-dimethylbiguanide in the formulation of the present invention is 20%≦C≦50%, preferably 25%≦C≦50%, preferably 30%≦C≦50%, more preferably 40%≦C≦50%, and particularly preferably 45%≦C≦50%.

[0235] In this invention, the indications (Y, X) are suitable for treating (Y) lesions or lesion symptoms (X), wherein X is the volume of the lesion, Y is an effective or rapid reduction, and wherein the lesion includes malignant lesions and non-malignant lesions.

[0236] In this invention, X represents the volume of the lesion, and Y represents an effective or rapid reduction. In this invention, the lesion is a lesion for which standard treatment is ineffective or has failed, including at least one selected from the group consisting of: tumors for which standard treatment is ineffective or has failed; malignant tumors refractory to standard chemotherapy drugs; and non-malignant nodules. Preferably, the lesion is a malignant tumor refractory to standard chemotherapy drugs.

[0237] In this invention, the refractory malignant tumor includes or is selected from at least one of the following groups: chemotherapy-resistant tumors, tumors with unfavorable microenvironments, tumors that have been discontinued, tumors for which there are no effective chemotherapy drugs, and tumors carried by patients who have contraindications or incompatibilities to standard chemotherapy drugs.

[0238] The non-malignant nodules are nodules containing at least one of the following non-malignant cells: connective tissue cells, secretory gland cells, and epithelial cells. In one embodiment, the nodules include or are selected from, for example: organ nodules rich in organ cells (e.g., breast nodules, thyroid nodules, pulmonary nodules), non-malignant tumors rich in connective tissue cells (lipomas, fibromas, etc.), connective tissue lesions rich in connective tissue cells (rheumatoid arthritis, systemic lupus erythematosus, polymyositis), and other nodules, preferably breast hyperplasia nodules.

[0239] In this invention, the lesion includes a tumor composed of non-drug-resistant malignant tumor cells.

[0240] In this invention, the malignant tumor cells are non-metformin cytotoxicity-specific tumor cells.

[0241] In this invention, the chemotherapy-resistant tumors include tumors resistant to standard chemotherapy drugs, including but not limited to those resistant to paclitaxel, 5-fluorouracil, cyclophosphamide, doxorubicin, and / or cisplatin, or those resistant to targeted antitumor drugs such as gefitinib, osimertinib, and / or trastuzumab. Examples include liver cancer tumors, breast cancer tumors, sarcoma tumors, pancreatic cancer, lung cancer, gastric cancer, and colon cancer tumors with unfavorable microenvironments. In one embodiment, the refractory malignant tumors include or are selected from chemotherapy-resistant liver cancer tumors, breast cancer tumors, sarcoma tumors, pancreatic cancer, lung cancer, gastric cancer, and colon cancer tumors. Preferably, the chemotherapy-resistant tumor is a 5-fluorouracil-resistant liver cancer.

[0242] In the above uses and methods of the present invention, the lesion includes tumors due to any pathology (malignant and / or non-malignant) and at any stage, including, for example, the following groups classified according to tumor cell type: epithelial cell tumors, sarcomas, lymphomas, germ cell tumors, germ cell tumors; and tumors named according to the organ or tissue in which the tumor cell concentration area is located, including, for example, tumors named according to the following organs or tissues: skin, bone, muscle, breast, kidney, liver, lung, gallbladder, pancreas, brain, esophagus, bladder, large intestine, small intestine, spleen, stomach, prostate, testicle, ovary, or uterus.

[0243] Specifically, the malignant tumors include, for example, breast cancer, pancreatic cancer, thyroid cancer, nasopharyngeal cancer, prostate cancer, liver cancer, lung cancer, intestinal cancer, oral cancer, esophageal cancer, stomach cancer, laryngeal cancer, testicular cancer, vaginal cancer, uterine cancer, ovarian cancer, sarcoma, etc.

[0244] Furthermore, the aforementioned malignant tumors include refractory tumors, such as tumors that have progressed after chemotherapy. Tumors that have progressed after chemotherapy include those that have progressed after treatment with chemotherapeutic agents such as paclitaxel, 5-fluorouracil, cyclophosphamide, doxorubicin, and cisplatin, or those that have progressed after treatment with targeted antitumor agents such as gefitinib, osimertinib, trastuzumab, and their derivatives.

[0245] 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, stomach tumors, nasopharyngeal tumors, laryngeal tumors, testicular tumors, vaginal tumors, uterine tumors, fallopian tube tumors, ovarian tumors, etc.

[0246] In the above uses and methods of the present invention, according to one embodiment, the malignant cells include or are selected from standard malignant cells that are ineffective alone by a standard metformin preparation, such as cancer cells other than liver cancer cells and sarcoma cells.

[0247] In the above uses and methods of the present invention, according to one embodiment, the non-malignant cells include or are selected from at least one of connective tissue cells and organ tissue cells.

[0248] In the above-described uses and methods of the present invention, according to one embodiment, the connective tissue cells include fibroblasts.

[0249] In the above-described uses and methods of the present invention, according to one embodiment, the organ tissue cells include or are selected from at least one of the following: hepatocytes, mammary gland cells, thyroid cells, and skin cells.

[0250] In the above uses and methods of the present invention, the non-malignant lesion includes or is selected from at least one of the following groups: benign tumors, nodules.

[0251] In the above-described uses and methods of the present invention, the non-malignant nodules include benign lesions other than benign tumors, including, for example, hyperplasia (e.g., hyperplasia of the breast, thyroid, parathyroid glands, prostate, etc.), cysts, abnormal venous masses (e.g., hemorrhoids), local inflammatory swelling, and swelling due to microbial infection. Hemorrhoids include internal hemorrhoids, external hemorrhoids, and mixed hemorrhoids.

[0252] In one embodiment, the benign lesion includes localized inflammation, particularly refractory inflammation. The refractory inflammation includes, but is not limited to, alterative inflammation, exudative inflammation, and proliferative inflammation, which can be any suitable type known to those skilled in the art, such as one or more of the following: arthritis, mastitis, pancreatitis, thyroiditis, prostatitis, hepatitis, pneumonia, enteritis, stomatitis, pharyngitis, periodontitis, esophagitis, gastritis, gastric ulcer, rhinitis, sinusitis, laryngitis, tracheitis, bronchitis, vaginitis, endometritis, salpingitis, oophoritis, etc.

[0253] In one embodiment, the benign lesion includes skin diseases, particularly refractory skin diseases. The refractory skin diseases include, but are not limited to, one or more of the following: skin cancer, non-malignant tumors of the skin, 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 tinea), sexually transmitted diseases (e.g., syphilis, gonorrhea, and condyloma acuminata), allergic and autoimmune skin diseases (e.g., contact dermatitis, eczema, urticaria), physical skin diseases (e.g., photodermatitis, chilblains, corns, chapped hands and feet, pressure sores), connective tissue diseases (e.g., lupus erythematosus), pigmentary disorders (e.g., freckles, nevi, various spots), diseases of skin appendages (e.g., acne, rosacea, seborrheic dermatitis, alopecia areata, hair loss, hyperhidrosis, and bromhidrosis), and cortical hyperplasia, such as spontaneous cortical hyperplasia.

[0254] In the above-described uses and methods of the present invention, the subject includes a mammal. Preferably, the subject includes a human. More preferably, the subject includes non-human mammals, such as non-human primates, livestock, and pets, including but not limited to, camels, pigs, cattle, sheep, horses, donkeys, goats, and sheep; and pets include various mammal pets such as dogs, cats, rabbits, monkeys, and orangutans.

[0255] In some embodiments, in the above-described formulations, pharmaceutical combinations, compositions, uses, methods, or kits, the intervention or administration concentration (C or CR, w / w) of the N,N-dimethylbiguanide is 1.0% ≦C ≦20%, preferably 3.0% ≦C ≦20%, more preferably 5.0% ≦C ≦20%, and particularly preferably 6.0% ≦C ≦20%.

[0256] In some embodiments, in the above-described formulations, drug combinations, compositions, uses, methods, or kits, the intervention or administration concentration (C or CR, w / w) of the N,N-dimethylbiguanide is 20%≦C≦50%, preferably 30%≦C≦50%, more preferably 40%≦C≦50%, and particularly preferably 45%≦C≦50%.

[0257] In some embodiments, in the above-described formulations, drug combinations, compositions, uses, methods, or kits, the tumor is a malignant tumor refractory to standard chemotherapy drugs. Preferably, the refractory malignant tumor includes or is selected from at least one of the following groups: chemotherapy-resistant tumors, tumors with unfavorable microenvironments, tumors that have been discontinued, tumors for which there are no effective chemotherapy drugs, and tumors carried by patients with contraindications or incompatibilities to standard chemotherapy drugs.

[0258] In some embodiments, in the above-described formulations, drug combinations, compositions, uses, methods, or kits, the nodule is a nodule comprising at least one of the following non-malignant cells: connective tissue cells, secretory gland cells, and epithelial cells.

[0259] In some embodiments, in the above-described formulations, drug combinations, compositions, uses, methods, or kits, the lesion includes a tumor containing non-drug-resistant malignant tumor cells. Simple Explanation of the Diagram

[0260] The following detailed description of specific embodiments of the present invention will be better understood when read in conjunction with the accompanying drawings. Exemplary embodiments are shown in the drawings for the purpose of illustrating the invention. However, it should be understood that the invention is not limited to the exact arrangement and means of the embodiments shown in the drawings.

[0261] Figure 1 (AC): HE staining pathological analysis (100×) of tumor samples from nude mouse tumor models (X, Y, Z groups) 1 hour after administration of metformin hydrochloride, N,N-dimethylbiguanide, and 5-FU containing human hepatocellular carcinoma 5-FU-resistant cells (Bel-7409 / 5-FU). X: Administration of 1.3% metformin hydrochloride; Y: Administration of 0.5% N,N-dimethylbiguanide; Z: Administration of 0.25% 5-FU (w / w).

[0262] Figure 2 (AC): Enlarged views (400×) of each HE staining pattern in Figures 1A-C.

[0263] Figure 3: Tumor samples from nude mouse tumor models 1 hour (AG) after administration of metformin hydrochloride, N,N-dimethylbiguanide and 5-FU to the nude mouse tumor model containing human hepatocellular carcinoma 5-FU-resistant cells (Bel-7409 / 5-FU) in Example 1.1.

[0264] Figure 4: Treatment effect of spontaneous cortical hyperplasia beagle in Example 4.2 (A: lesion before treatment, B: lesion after treatment).

[0265] Figure 5: Treatment effect in rats with spontaneous fibroma in Example 4.2 (A: lesions before treatment, B: lesions after treatment). Implementation

[0266] [definition]

[0267] In this application, the term "activity" refers to the ability of a compound to exert a specific biological or pharmacological effect. The term "active form" refers to the specific form in which a compound provides a specific activity, such as the specific form that binds to a specific target and triggers a biological effect. The term "active ingredient" refers to a chemical entity in a drug or formulation that can directly provide a specific active form. The original state of a compound may not necessarily possess direct activity; for example, salicylic acid, produced by the hydrolysis of aspirin, is its active form, while sulfonamide drugs need to dissociate into a free state to exert their antibacterial effect.

[0268] In this application, the term "standard metformin" (denoted as R' in this application) refers to metformin as used in the prior art. This metformin, defined by its key characteristics (systemic administration and the ability to provide the desired pharmacological effects through systemic administration), can only be a metformin salt. For example, in the catalogs of major suppliers like Sigma, the only compound under the metformin category is dimethyl biguanide hydrochloride. Any metformin sourced from Sigma, or any metformin that can be used in cell experiments, or that can be administered systemically (including intraperitoneally), or that has the same mechanism of action as systemic administration, can only be a metformin salt. In this application, the most commonly used term "metformin," namely dimethyl biguanide hydrochloride, is used as a representative of the standard metformin drug. In the prior art, metformin salt and metformin are often used interchangeably.

[0269] In this application, the term "N,N-dimethylbiguanide" (designated R in this application, and can be used interchangeably with metformin compounds) is a distinguishing compound of standard metformin (metformin salts). According to the methods of the International Union of Pure and Applied Chemistry (IUPAC), a metformin compound called N,N-dimethylbiguanide has the chemical formula C4H11N5, the simplified structural formula H3CNCNH3CNCH3, and a molecular weight of 129.2 (see Formula 1 below for specific results). Metformin salts are salts formed by N,N-dimethylbiguanide with various acids, with the chemical formula C4H11N5 and the simplified structural formula H3CNCNH3CNCH3. The specific IUPAC designation depends on the acid contained within. For example, the salt formed by N,N-dimethylbiguanide and hydrochloric acid has the chemical formula C4H11N5.HCl and the simplified structural formula H3CNCNH3CNCH3.HCl. IUPAC names it N,N-dimethylbiguanide hydrochloride, with a molecular weight of 165.6 (see Formula 2 below).

[0270] Different IUPAC standard nomenclatures correspond to distinct chemical structures. In N,N-dimethylbiguanide, "N,N-dimethyl" indicates that the molecule has two methyl groups (CH3-) attached to two nitrogen atoms, one for each. "Biguanide" refers to a derivative of biguanide, where the guanidinium group is free. In N,N-dimethylbiguanide hydrochloride, the guanidinium group adsorbs a chloride ion, forming a positively charged biguanide ion and its corresponding chloride ion pair. Other biguanide salts follow the same pattern. This structural difference is also reflected in their different preparation methods. N,N-dimethylbiguanide hydrochloride is relatively inexpensive to prepare; N,N-dimethylbiguanide is typically prepared from N,N-dimethylbiguanide hydrochloride as a raw material.

[0271] In fact, when N,N-dimethylbiguanide enters bodily fluids such as blood, it can form metformin salts with acid radicals therein. For example, the guanidinyl group in N,N-dimethylbiguanide combines with chloride ions (Cl-) to form an ion pair, resulting in N,N-dimethylbiguanide hydrochloride. In the prior art, the aforementioned structural differences in vitro are not expected to produce significant differences in biological activity; R and R' are considered pharmaceutically equivalent.

[0272] However, the results from the embodiments of this application unexpectedly show that this structural difference produces distinct biological activities under specific conditions, i.e., distinct structure-activity relationships. Based on these results, it is possible to more comprehensively evaluate whether N,N-dimethylbiguanide and metformin salts are different active ingredients. Assuming they are both administered systemically, dispersed in the bloodstream in the same form, and then exert the same effect, they are pharmaceutically equivalent and considered the same active ingredient. However, if they are administered locally, have different active forms in a specific solution (e.g., aqueous solution), and these different forms provide different core activities and exert different effects, then they are not pharmaceutically equivalent and their active ingredients must be defined separately according to their different active forms.

[0273] In fact, N,N-dimethylbiguanide (R) under the conditions of this application has a morphological manifestation that differs from that of standard metformin (whose active ingredient is R'), wherein the active form of R' is a form in which it is completely complexed with acid in solution (shown in Example 5 as being suitable for systemic administration); conversely, the active form of R is a form in which it is less complexed with acid in solution, and is mainly present in a form of very little or partial complexation (shown in Example 5 as being unsuitable for systemic administration and only suitable for local administration), thereby providing an activity (activity A) that R' cannot provide, which includes transient local effects.

[0274] In this application, the term "cell-responsive drug" (designated B) refers to a class of anti-tumor drugs that target tumor cells, primarily by regulating tumor cell growth, division, or death signaling pathways and intervening in key metabolic and biological processes of tumor cells to achieve anti-tumor effects. This class of drugs includes cytotoxic drugs and targeted anti-tumor drugs. The term "cytotoxic drug" (designated B1) refers to a class of anti-tumor drugs that kill tumor cells through non-specific mechanisms (directly damaging DNA, inhibiting RNA synthesis, or interfering with protein polymerization). The term "targeted anti-tumor drug" (designated B2) refers to a class of drugs that selectively inhibit tumor cell growth or induce tumor cell death by binding to tumor cell-specific targets (such as specific gene mutation products or abnormally expressed receptor proteins) to regulate cell signaling pathways. While cytotoxic drugs and targeted anti-tumor drugs are generally considered very different, they share the following common characteristics: inhibiting tumor progression by intervening in key tumor cell signaling pathways (such as proliferation, metabolism, apoptosis, or DNA repair), thus facing a common challenge.

[0275] In this application, the term "near-neutral or weakly basic sodium salt" (abbreviated as C) refers to a sodium salt compound that can be used for injection and whose aqueous solution exhibits near-neutral pH (5.0-8.5) within a certain concentration range. Such sodium salts include, but are not limited to, sodium chloride, sodium bicarbonate, sodium dihydrogen phosphate, sodium lactate, sodium acetate, etc.

[0276] In this application, the term "immunomodulator" refers to substances that can optimize local and / or systemic immune responses by utilizing disease-related immune substances and / or inflammatory signals released within the lesion, including but not limited to the following three categories: biological macromolecular immunomodulators, immune cell-based immunomodulators, and vaccine-based immunomodulators. The term "biological macromolecular immunomodulator" (E1) refers to protein- or nucleic acid-based immunomodulators, with representative protein-based drugs being immune checkpoint inhibitors and cytokines, and representative nucleic acid-based drugs being TLR agonists. The term "immunocell-based immunomodulator" (E2) refers to cell products used in treatments that directly or indirectly enhance immune responses through immune cell transplantation or injection, such as DC vaccines, NK cells, and CAR-T cells. The term "vaccine-based immunomodulator" (E3) refers to preparations that promote immune responses by inducing specific or non-specific immune enhancement, such as disease-related immune substance vaccines, oncolytic virus vaccines, and BCG vaccines.

[0277] In this application, the term "formulation" refers to a pharmaceutical product that can be used directly, prepared from pharmaceutical raw materials through a specific manufacturing process. This product may be, for example, tablets, capsules, injections, creams, suspensions, etc. The term "optimized formulation" refers to a pharmaceutical preparation that, through systematic research and optimization evaluation, selects the most suitable pharmaceutical raw materials, excipients, and manufacturing processes to achieve optimal performance in terms of drug use (most preferably oral administration, followed by other systemic applications), efficacy, stability, and safety. After decades of selection, topical formulations are far from being the optimal formulations for standard metformin.

[0278] In this application, the term "regimen" refers to a specific combination of drugs, a pharmaceutical composition, a formulation, a method, a kit, or an application. The term "regimen of this application" refers to the R-based combination of drugs, a pharmaceutical composition, a formulation, a method, a kit, or an application of this application. The term "control regimen" refers to a combination of drugs, a pharmaceutical composition, a formulation, a method, a kit, or an application used as a control of the regimen of this application. The term "metformin standard regimen" refers to an R'-based regimen (commonly used as a control regimen). The indication is the core effect and core value of the regimen.

[0279] In this application, the term "indication" is distinguished from "disease or disease symptom," the latter referring to an abnormal life process, while the former refers to "a disease or condition suitable for a particular treatment" (Cihai (1989 edition)) or "the range of symptoms for which a drug is applicable" (National Science and Technology Terminology Committee, Pharmaceutical Terminology (Second Edition)). In short, the indication of a treatment regimen refers to the range of specific diseases or disease symptoms for which it is applicable, based on its efficacy characteristics (Y), to address the clinical needs of a specific disease (X) or its core pathological conditions, denoted as (X,Y). Those skilled in the art know that efficacy characteristics (Y) are the core condition limiting the range of applicable diseases or symptoms (X), and must be specific, rigorous, clear, and error-free; authoritative bodies in various countries have regulations on this. For example, the China Food and Drug Administration (CFDA) stipulates in the "Detailed Rules for the Specification of Instructions for Chemical Drugs and Therapeutic Biological Products (2006 Edition)" that "[Indication] should be expressed accurately according to the purpose of the drug, clearly stating that it is used for the prevention, treatment, diagnosis, relief, or adjunctive treatment of a certain disease (state) or syndrome." The efficacy characteristics (Y) of a drug or method determine its purpose, such as Y = prevention, treatment, diagnosis, relief, or adjuvant treatment. In the field of "treatment", Y can be further subdivided according to the more refined efficacy characteristics it meets. For example, based on the satisfaction of a specific effect, it can be distinguished as: effective treatment, rapid-acting treatment, highly effective treatment, long-acting treatment, and heterogeneous equivalence (regimen).

[0280] In this application, the term "structure-activity relationship" refers to the association between an indication (X, Y) and a set of drug components, such as (α, β, γ). An indication is the most essential and valuable set of characteristic drug "efficacy" of a pharmaceutical regimen, and the set of necessary components is obviously its characteristic drug "structure." The pharmacological conditions that enable the drug "structure" to exhibit drug "efficacy" define the relationship between them, i.e., f: (α, β, γ) → (X, Y). Wherein, Y represents the efficacy characteristics, including synergy, dose-response, therapeutic efficacy (time-effect, effectiveness, long-term effect), and heterogeneous adaptability; X represents the range of lesions for which efficacy characteristic Y is suitable for application; α represents the medicinal chemical characteristics or set of efficacy characteristics Y required to satisfy indication X, such as a common chemical structure subset (A, B); β represents the conditions that provide activity for the active ingredient in the drug, such as the dosage of A and the functional combination or synergistic conditions of (A, B), including, for example, the common concentration thresholds of A and B and their synergistic concentration ratio range, etc.; γ represents formulation compatibility (the exclusivity of A, etc.), ensuring the stability and efficacy of the drug under specific conditions. The relation f maps the "structure" set (α, β, γ) to the "effect" set (X, Y), f: (α, β, γ) → (X, Y), f including, for example, the administration method of the active form of the structure required to form the effect.

[0281] As described in the "Technical Background" section, the standard metformin regimen has an EC50 in the mM range in sensitive tumor cell experiments, which is more than 100 times higher than that of standard antitumor drugs (e.g., 5-FU). Aside from safety, it offers no applicable advantages in activity or efficacy characteristics (Y). For example, the indications for the standard metformin regimen are limited to (X' = sensitive tumors, Y' = effective but not rapid-acting, highly effective, or long-acting treatment), rather than the commonly expanded claims of (X = tumor, Y = treatment). Extending its indications (X', Y') to (X, Y) could have serious clinical consequences due to misuse of the regimen, delaying treatment and misleading those skilled in the art.

[0282] The scientific combination of α, β, and γ is key to achieving the desired effect of a drug, reflecting a precise match to a specific indication (X, Y). For example, differences in (α, β, γ) between R and R' include: different molecular structures (α) providing the active form (N,N-dimethylbiguanide vs. N,N-dimethylbiguanide hydrochloride), different concentration thresholds or dosage solubility (β) (QR / QR' < 50%), and different drug formulation stability (γ) (with vs. without mixing contraindications).

[0283] This application provides a new set of drug components (α, β, γ) and a new pharmacology (f) to generate new therapeutic properties or (Y) and the diseases or conditions (X) suitable for treatment, i.e., f:(α, β, γ)→(X,Y).

[0284] The comparison system, evaluation methods and standards, and related definitions of the effect characteristics (Y) are as follows.

[0285] 1. Comparison System

[0286] In this application, the term "standard comparison" refers to comparing the research proposal with existing or technically predicted solutions (standard solutions) to assess whether the research proposal's technical effectiveness exceeds the expectations of the standard solution; the term "further comparison" refers to conducting in-depth evaluations of multiple research proposals based on the standard comparison, and further evaluating whether a combination of proposals exceeds the expectations based on the component proposals through refined and expanded comparisons.

[0287] Standard comparisons of single-drug therapy primarily involve comparing metformin (R) with a standard metformin drug (R'). According to existing technology, there are no structural differences between metformin drugs; R and R' are considered to have the same active ingredient and are expected to have the same efficacy. The actual efficacy of R in the study is compared with the results of R'. If the efficacy is the same, it meets expectations; if the efficacy is significantly stronger, it exceeds expectations and becomes a non-standard drug that conflicts with the aforementioned teachings.

[0288] The standard comparison of combinations mainly involves comparing the combination R / Z with the standard combination R' / Z. As mentioned above, based on the teachings of the prior art, it is expected that R / Z and R' / Z have the same efficacy. The actual efficacy of R / Z in the research protocol is compared with the results of R' / Z. If the efficacy is the same, it meets the expectation; if the efficacy is significantly stronger, it exceeds the expectation and becomes a super-standard combination that conflicts with the aforementioned teachings.

[0289] Given that R becomes the off-standard metformin drug, the comparison between the study combination R / Z and its individual drugs, especially R, clearly exceeds the expected range of standard comparisons. Standard comparisons are no longer applicable, and further comparisons must be made between different study protocols. Further comparisons are based on the HSA or Bliss method described below for single-drug results to predict the combination's outcome. The actual efficacy of R / Z in the study protocol is compared with the expected efficacy; if the efficacy is the same, it meets expectations; if the efficacy is stronger, it exceeds expectations, becoming a further off-standard combination.

[0290] 2. Interaction

[0291] The expected interactions (antagonistic, additive, synergistic) between the active components (e.g., R and Z) in a shared scheme are highly uncertain; this is known as synergy. The probability of synergy is much lower than that of non-synergy. Its core is synergy that exceeds the expectations of existing technical solutions. This is usually an important objective of shared scheme research, as well as a major technical challenge and essential characteristic. It also provides important evidence of unique innovation in the combinatorial mechanism.

[0292] In this application, taking the mouse experiment in the examples as an example, the evaluation of compatibility is as follows:

[0293] In this application, the dose-response evaluation is carried out using mouse experiments as an example. (1). Calculate the compatibility index The compatibility index is the ratio of the actual sharing effect (q1) widely used in the industry to the expected sharing effect (q2) calculated based on the prediction model: q = q1 / q2. (2). General evaluation When q>1, it is synergistic; otherwise, it is non-synergistic (it may be an independent or antagonistic effect). (3) Relative evaluation (A) Standard Comparison In the standard comparison, q2 represents the expected shared effect of R / Z based on existing technology (R / Z and R' / Z are equivalent). The evaluation criterion is: if the actual effect of R / Z (q1) is greater than R' / Z, then q = q1 / q2 > 1.0 indicates synergy; otherwise, it indicates non-synergy.

[0294] The unexpected synergistic effect of R replacing R' (metformin in the prior art) in combination with Z manifests as an unexpected synergistic effect. For example, the combination of standard metformin (R') and a cell-responsive drug (B) is expected to result in an interaction between cellular effects; however, the combination of N,N-dimethylbiguanide (R) and B in this application provides an interaction between the weakening effect of R on the local biological barrier function and the cell-responsive antitumor effect of B, the latter exceeding the former's expectation. This is the aforementioned synergistic, out-of-standard pharmacological mechanism.

[0295] (B) Further comparison

[0296] In standard comparisons where the R / Z combination exhibits synergy, further comparisons are conducted. q2 represents the expected combined effect based on the efficacy of individual drugs R and Z, where R represents the expected active component beyond the existing metformin technology. The calculation of q2 compares the combination regimen with the single-drug regimen and can be used for both standard and further comparisons. The two most commonly used methods in antitumor animal studies are selected below. In this application, the actual single-drug effects of drugs R and Z are denoted as ER and EZ, respectively.

[0297] (a) Highest Single-Drug Effect (HSA) Model

[0298] The Highest Single Agent (HSA) method is one of the most widely used methods in antitumor animal experiments. Its core idea is to compare the actual effect of combined drug therapy (q1) with the predicted value of the best single-drug effect (q2), i.e., q2 = max(ER, EZ). If the effects are the same, it meets expectations; if the effect is stronger, it exceeds expectations. The evaluation criterion is: if the actual efficacy of the combination (q1) is greater than max(ER, EZ), it is considered further synergistic; otherwise, it is not considered further synergistic.

[0299] (B). Bliss Method

[0300] The Bliss independence model (Bliss, CI (1939). The toxicity of poisons applied jointly. Annals of Applied Biology, 26(3), 585-615.) is another widely used method in antitumor animal experiments. Its core assumption is that the effects of the two drugs are independent of each other and follow the principle of probability superposition: q2 = ER + EZ - ER × EZ. If the effects are the same, it is in line with expectations; if the effects are stronger, it exceeds expectations. The evaluation criterion is: if the actual effect of the combination (q1) is greater than q2, it is considered further synergy; otherwise, it is not considered further synergy.

[0301] In this application, the terms "further synergy" (scheme), "synergy" (scheme), and "non-synergy" (scheme) refer to (schemes) that meet the above evaluation criteria for further synergy, synergy, and non-synergy, respectively.

[0302] In one embodiment, this application provides a drug combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, wherein the combination of R and Z (R / Z combination) is a synergistic combination that exceeds the expected effect of a standard metformin drug (R') and Z combination (R' / Z combination), characterized by a significant increase in drug effect when the dosage of R (QR) is ≤25% or 50% of the dosage of R (QR') and the dosage of Z is not increased, including an increase of at least one drug adjustability indicator by at least 100%, an increase of at least one drug efficacy indicator by at least 25%, or an increase in local efficacy-toxicity ratio by at least 25%.

[0303] In one embodiment, this application provides a combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, wherein the combination of R and Z (R / Z combination) is a further synergistic combination exceeding the expected effects of the HSA or Bliss method based on R monotherapy and Z monotherapy, characterized by an increased drug effect when the dosage of R (QR) is not increased and the dosage of Z (QB) is reduced by at least 20%, including an increase of at least one drug adjustability indicator by at least 10%, an enhancement of at least one drug efficacy indicator by at least 10%, or an increase in the local efficacy-to-toxicity ratio by at least 10%.

[0304] In one embodiment, the synergistic scheme or further synergistic scheme of this application is preferably applicable to tumors that require the interaction between the weakening effect of R on the tumor barrier function and the intratumoral cellular reactive antitumor effect of B to produce higher efficacy, including, for example, highly aggressive tumors, advanced tumors, rapidly deteriorating tumors, and refractory tumors.

[0305] 3. Dose-effectiveness

[0306] In this application, the term "dose-effectiveness" refers to the strength of the effect per unit dose of a drug. The expected dose-effectiveness of a regimen can range from potent to weak to ineffective, exhibiting a high degree of uncertainty. The difference between normal cells and tumor cells is not significant, and the effective dose and safety limit of antitumor drugs are usually not far apart, resulting in low expected efficacy and manifesting as severe side effects even when patient benefit is minimal. Improving dose-effectiveness is a crucial objective of regimen research, as well as a major technical challenge and essential characteristic. If a new regimen exceeds the expected dose-effectiveness of an older regimen, it not only demonstrates unexpected synergy but also provides important evidence of unique innovation in treatment mechanisms or drug design.

[0307] In this application, the dose-response evaluation is carried out using mouse experiments as an example.

[0308] (1) Quantity-effectiveness indicators

[0309] In this application, the term "dose-effectiveness index" refers to the numerical representation of a specific drug effect (such as therapeutic effect) produced in the body by each unit of drug (such as per mmol or per milligram of drug), used to measure the minimum dosage of a drug or the relationship between its therapeutic effect (X) and the minimum dosage or the minimum frequency of administration associated with it, usually the number of treatment courses.

[0310] (2). Relative evaluation

[0311] (A) Standard Comparison

[0312] Monotherapy comparison: The evaluation criteria are as follows: when the dosage of R is halved (QR=1 / 2QR'), at least one of its drug effects is increased by at least 30%; or at least one of the actual dose-response indicators of R monotherapy is increased by at least 50% or 70% compared with R' monotherapy, then its dose-response effect exceeds the expectations of the prior art (R and R' are regarded as pharmaceutical equivalents) and is considered to be potent.

[0313] Combination comparison: The evaluation criteria are as follows: when the dosage of at least one of the R / Z components is halved, the pharmacological effect of at least one component of the combination is increased by at least 30%; or the actual dose-response index of at least one of the R / Z components is increased by at least 50% or 70% compared with the corresponding component in R' / B, then the dose-response of the combination exceeds the expectations of the prior art (R and R' are pharmaceutical equivalents) and is considered potent.

[0314] (B) Further comparison

[0315] In further comparisons, the R / Z ratio is determined by the lowest common dose-response indices of the single-drug R and Z (HSA method). The evaluation criteria are: when R / Z is potent, and when the dosage of at least one of R and Z is reduced by 10%, the actual dose-response indices of the combination are improved by at least 10% or 20% compared to the highest of the single-drug ratios, then its dose-response performance is further enhanced to be considered potent beyond expectations.

[0316] In this application, the terms "further enhanced" (scheme), "enhanced" (scheme), and "non-enhanced" (scheme) refer to (schemes) that meet the above evaluation criteria for further enhanced, enhanced, and non-enhanced effects, respectively.

[0317] In one implementation, the differential structure-activity relationship of the present application can be simply expressed as f: (α, β, γ) → (X, Y), wherein the differential indication (X, Y) is the range (X) to which the potent or further potent R or R / Z is applicable, while the non-potent R' or R' / Z is not applicable: patients with local lesions who need to benefit from high efficacy at low doses or low frequencies; the differential pharmacology (f) mainly includes the transient local effects of the potent or further potent R that distinguish it from R'; the differential component set (α, β, γ) is the selection and range of potent or further potent components defined by (X, Y) and f.

[0318] In one embodiment, this application provides a formulation, method, kit, or use for treating local lesions, wherein R is a potent active ingredient that exceeds the expected efficacy of a standard metformin drug (R'), characterized by a significant increase in drug effect when the R-course dosage (QR) is ≤25% or 50% of the R'-course dosage (QR'), including an increase of at least one drug adjustability indicator by at least 100%, an increase of at least one drug efficacy indicator by at least 25%, or an increase in the local efficacy-toxicity ratio by at least 25%.

[0319] In one embodiment, this application provides a combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, wherein the combination of R and Z (R / Z combination) is a potent combination that exceeds the expected efficacy of a standard combination of metformin (R') and Z (R' / Z combination), characterized by a significant increase in drug effect when the dosage of R (QR) is ≤25% or 50% of the dosage of R' (QR') and the dosage of Z is not increased, including an increase of at least one drug adjustability indicator by at least 100%, an increase of at least one drug efficacy indicator by at least 25%, or an increase in local efficacy-to-toxicity ratio by at least 25%.

[0320] In one embodiment, this application provides a further dose-reduction and synergistic combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, wherein the combination of R and Z (R / Z combination) is a further synergistic combination that exceeds the expected effects of the HSA or Bliss method based on R monotherapy and Z monotherapy, characterized by an increased drug effect when the dosage of R (QR) is not increased and the dosage of Z (QZ) is reduced by at least 20%, including an increase of at least one drug adjustability indicator by at least 10% and an enhancement of at least one drug efficacy indicator by at least 10%.

[0321] In one embodiment, the potent or further potent regimens described in this application are preferably suitable for tumors requiring the strongest possible dose-response for treatment, including, for example, highly aggressive tumors, advanced tumors, rapidly worsening tumors, and refractory tumors. If the aforementioned control regimen is chosen instead of the regimen described in this application, there is a significant clinical risk of increased drug dosage due to misuse of the regimen (selecting a non-potent regimen when a potent one should be preferred), resulting in low efficacy and missed treatment opportunities.

[0322] 4. Therapeutic effect

[0323] In this application, the term "therapeutic efficacy" refers to the ability to produce a significant therapeutic effect on a target disease or symptom, including, for example, time-dependent, effective, and long-lasting effects. The level of therapeutic efficacy is influenced by various factors and is highly uncertain (high efficacy, low efficacy, no efficacy, etc.). High efficacy has the lowest probability of occurrence and is a major technical challenge in drug development, as well as an important fundamental indicator of innovative treatment regimens. If a new regimen exceeds expectations by being fast-acting, effective, or long-lasting when the efficacy of an existing regimen is not expected (e.g., non-rapid-acting, non-effective, or non-durable), this not only constitutes unexpected synergy but also provides important evidence of unique innovation in the treatment mechanism or drug design.

[0324] 1) Timeliness

[0325] In this application, the term "timeliness" refers to the length of time it takes for a treatment regimen to show an effect (significant effect) on a target disease or symptom, which is an important indicator for measuring the speed of treatment effectiveness. The expected timeliness of a treatment regimen includes rapid-acting, non-rapid-acting, etc., and is highly uncertain. Cancer typically progresses rapidly, and the expected timeliness (e.g., anti-tumor drugs) is usually not high. The probability of rapid-acting effects is far less than that of non-rapid-acting effects; this is an important objective of regimen research, as well as a major technical challenge and essential characteristic. If a new regimen exceeds the expected rapid-acting effect of an older, non-rapid-acting regimen, it not only represents unexpected synergy but also provides important evidence of unique innovation in the combination mechanism.

[0326] In this application, taking the mouse experiment in the examples as an example, the timeliness evaluation is as follows.

[0327] (1) Timeliness indicators

[0328] In this application, the term "timeliness index" refers to the time to significant effect, wherein significant effect means a relative tumor proliferation rate (T / C%) ≤ 42% (according to relevant NCI guidelines), and the time to significant effect is the date on which significant effect is observed.

[0329] (2). Relative evaluation

[0330] (A) Standard Comparison

[0331] Monotherapy comparison: The evaluation criteria are: when the actual effective time of monotherapy R is at least 25% or 40% shorter than that of monotherapy R', its timeliness exceeds the expectations of the existing technology (R and R' are pharmaceutical equivalents) and is considered rapid-acting.

[0332] Combination comparison: The evaluation criteria are: when the actual effective time of R / Z is shorter than that of R' / Z, or at least 25% or 40% shorter, then its timeliness exceeds the expectations of the existing technology (R / Z and R' / Z are pharmaceutical equivalents) and is considered to be fast-acting.

[0333] (B) Further comparison

[0334] In further comparisons, R / Z was determined to be the fastest among the single-drug drugs R and Z based on their expected time-to-action (HSA method). The evaluation criterion was: if the actual onset time of R / Z was shorter than that of the fastest single-drug drug R and Z, or at least 10% or 20% shorter, then its time-to-action performance exceeded expectations, constituting a further rapid effect.

[0335] In this application, the terms "further rapid-acting" (regimen), "rapid-acting" (regimen), and "non-rapid-acting" (regimen) refer to (regimens) that meet the above evaluation criteria for further rapid-acting, rapid-acting, and non-rapid-acting, respectively. Drugs with strong time-efficacy can be better combined with other treatment modalities, enhancing combination potential.

[0336] In one embodiment, the differential structure-activity relationship of the present application can be simply expressed as f: (α, β, γ) → (X, Y), wherein the differential indication (X, Y) is the range (X) to which the rapid or further rapid-acting (Y) effect of R or R / Z is applicable, while the non-rapid-acting (X) effect of R' or R' / Z is not applicable: patients with local lesions requiring rapid response during disease progression; the differential pharmacology (f) mainly includes the transient local effects of R that are different from R's rapid or further rapid-acting effects; the differential component set (α, β, γ) is the selection and range of rapid or further rapid-acting components defined by (X, Y) and f.

[0337] In one embodiment, this application provides a formulation, method, kit, or use for treating local lesions, wherein R is a reduced-dose, fast-acting active ingredient that exceeds the expected effect of a standard metformin drug (R'), characterized by a significantly increased drug effect when the R-course dosage (QR) is ≤25% or 50% of the R'-course dosage (QR'), including a significantly improved drug efficacy index, as evidenced by a reduction of at least 25% in the time to efficacy (the day when tumor proliferation rate <42%) compared to said R' in mouse experiments.

[0338] In one embodiment, this application provides a combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, wherein the combination of R and Z (R / Z combination) is a reduced-dose, rapid-acting combination that exceeds the expected efficacy of a standard metformin combination (R') and Z combination (R' / Z combination). This is manifested in a significantly increased drug effect when the R-course dose (QR) is ≤25% or 50%, the R'-course dose (QR') and the Z-course dose are not increased, including a significantly improved drug efficacy index, manifested in a reduction of at least 25% in the time to efficacy (the day when tumor proliferation rate <42%) compared to the R' / Z combination in mouse trials.

[0339] In one embodiment, this application provides a further dose-reduction and synergistic combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, wherein the combination of R and Z (R / Z combination) is a further dose-reduction rapid-acting combination that exceeds the expected efficacy of R monotherapy and Z monotherapy using the HSA or Bliss method, characterized by a significant increase in drug effect when the dose of R for one course of treatment (QR) is not increased and the dose of Z for one course of treatment (QZ) is reduced by at least 20%, including a significant improvement in drug efficacy indicators, as demonstrated by a reduction of at least 25% in the time to onset of action (the day when the tumor proliferation rate is <42%) compared to R' in mouse studies.

[0340] In one embodiment, the dose-reduction rapid-acting regimen of this application is preferably applicable to tumors requiring the strongest possible dose-response and the fastest possible rapid-acting regimen, including, for example, highly aggressive tumors, advanced tumors, rapidly worsening tumors, refractory tumors, and tumors with urgent symptoms requiring rapid defecation. If the standard metformin regimen is chosen instead of the potent rapid-acting regimen of this application, there may be numerous risks in clinical practice due to misuse of the regimen, resulting in increased drug dosage but low efficacy, prolonged time to onset of action, and thus missed treatment opportunities.

[0341] 2) Effectiveness

[0342] In this application, the term "efficacy" refers to the ability to suppress a target disease or symptom. The expected efficacy of a regimen can be highly uncertain, ranging from highly effective (effective but not highly effective), ineffective but with some therapeutic effect (ineffective but with some therapeutic effect), to unexpected efficacy (ineffective and with no reasonably expected therapeutic effect). Expected efficacy is usually low; for example, in anti-tumor drugs, the expected efficacy of first-line treatments is usually highly effective, while second-line treatments are mostly ineffective, and those still available outside of first and second-line treatments are almost ineffective but with some therapeutic effect (e.g., adjuvant therapy), with most drugs actually having no expected efficacy. Therefore, the probability of highly effective treatment is far less than that of ineffective but with some therapeutic effect, and the latter is far less than that of ineffective but with some therapeutic effect. Improving efficacy is a key objective of regimen research, as well as a major technical challenge and essential characteristic. If a new regimen exceeds the expected efficacy of an old regimen, this not only provides evidence of unexpected synergy but also crucial evidence of unique innovation in the treatment mechanism or drug design.

[0343] In this application, the effectiveness is evaluated using mouse experiments as an example.

[0344] (1) Calculate the effectiveness index

[0345] In this application, the term "efficacy endpoint" refers to tumor inhibition rate or objective response rate. When a negative control group is available, the efficacy endpoint is tumor inhibition rate and tumor weight; when no negative control group is available, the efficacy endpoint is the objective response rate (ORR). The latter classifies the efficacy of the study group into the following four levels (RECIST guidelines): complete response (CR) – disappearance of target lesions; partial response (PR) – a 30% reduction in target lesion volume; disease progression (PD) – an increase in target lesion volume of 20% or more; and stable disease (SE) – a change in target lesion volume between PR and PD. ORR is the sum of CR and PR within a predetermined minimum timeframe (e.g., N days after drug discontinuation).

[0346] (2). General evaluation

[0347] When a negative control group is present, the efficacy evaluation criteria for the study group are as follows (CFDA's "Guidelines for Pharmacodynamics of Antitumor Drugs"): tumor inhibition rate. A tumor weight difference of 40% compared to the negative control group was statistically significant (P < 0.05) to be considered effective; otherwise, it was considered ineffective. In the absence of a negative control group, the commonly used criterion for evaluating effectiveness is: an ORR ≥ 60% is considered effective; otherwise, it is considered ineffective.

[0348] For ineffective study groups, the evaluation criteria were: when the tumor inhibition rate was >25% or the objective response rate was >40%, it was considered ineffective and useful; otherwise, there was no expectation of (efficacy).

[0349] (3) Relative evaluation

[0350] (A) Standard Comparison

[0351] Monotherapy comparison: The evaluation criteria are: when the actual tumor inhibition rate or objective response rate of R monotherapy is at least 25% or 50% higher than that of R' monotherapy, its efficacy exceeds the expectations of existing technology (R and R' are pharmaceutical equivalents) and is considered highly effective.

[0352] Combination comparison: The evaluation criteria are: when the actual tumor inhibition rate or objective response rate of R / Z is at least 10%, 20%, or 40% higher than that of R' / Z, its efficacy exceeds the expectations of existing technologies (R / Z and R' / Z are pharmaceutical equivalents) and is considered highly effective.

[0353] (B) Further comparison

[0354] In further comparisons, the R / Z ratio was determined by the combined efficacy expectation of R and Z monotherapy (HSA method), which was the highest among them. The evaluation criteria were: when R / Z was highly effective, and its actual tumor inhibition rate or objective response rate was at least 10% or 20% higher than the highest of R and Z monotherapy, its efficacy exceeded expectations and was considered further highly effective.

[0355] In this application, the terms "further efficient" (scheme), "efficient" (scheme), "ineffective" (scheme), "ineffective useful" (scheme), and "unexpected" (scheme) respectively refer to (schemes) that meet the above evaluation criteria of further efficient, efficient, ineffective, ineffective useful, and unexpected.

[0356] In this application, the term "refractory tumor" refers to a tumor for which a particular regimen, such as a cell-responsive drug or standard metformin, is not effective, including tumors in patients who are contraindicated or incompatible with the regimen and tumors that are not effective indications for treatment.

[0357] In this application, the term "non-indicative tumor" refers to a tumor that is not included in the scope of a particular regimen, such as a cell-responsive drug or a standard metformin drug. For example, tumors that are not included in the list of drugs published by an authoritative institution as first- or second-line treatments include ineffective useful tumors and hopeless tumors. Ineffective useful tumors include, for example, tumors with unfavorable microenvironment, and hopeless tumors include, for example, drug-resistant tumors and discontinued tumors.

[0358] In this application, the term "drug-resistant tumor" refers to a tumor containing drug-resistant cells or tissues (e.g., tumor tissue derived from patients resistant to clinical anti-tumor drugs or targeted therapies). Drug-resistant tumors are also usually ineffective against a variety of other anti-tumor drugs, thus becoming refractory tumors.

[0359] In this application, the term "microenvironmentally unfavorable tumor" refers to a tumor containing a microenvironment that is unfavorable to the action of antitumor drugs, such as tumors formed by a mixture of tumor cells and fibroblasts in animal experiments, or tumors derived from clinically high-stromal-to-interstitial-to-tumor tissues (e.g., pancreatic cancer tissue). Such microenvironments, according to known principles, significantly reduce the sensitivity of tumor cells within them to standard antitumor drugs, thus making them refractory tumors.

[0360] In this application, the term "discontinued tumor" refers to a tumor that has been discontinued from antitumor drugs for any reason. There are many reasons for discontinuing antitumor medication, including unsuitability of the patient's physical condition and drug treatment failure, which in turn includes drug desensitization for any reason. The mechanisms of this desensitization are numerous and not fully elucidated by current research, involving both cellular (e.g., cellular drug resistance) and tissue (e.g., increased difficulty in tissue penetration), thus becoming refractory tumors.

[0361] In one embodiment, the differential structure-activity relationship of the present application can be simply expressed as f: (α, β, γ) → (X, Y), wherein the differential indication (X, Y) is the range (X) to which the highly effective or further highly effective (Y) R or R / Z is applicable, while the ineffective, ineffective useful, or not expected to be applicable (X) R' or R' / Z is: patients with local lesions requiring highly effective or further highly effective treatment; the differential pharmacology (f) mainly includes the transient local effects of the highly effective or further highly effective R compared to R'; the differential component set (α, β, γ) is the selection and range of highly effective or further highly effective components defined by (X, Y) and f, wherein X includes cell-reactive drugs or tumors refractory to standard metformin drugs.

[0362] In one embodiment, this application provides a formulation, method, kit, or use for treating local lesions, wherein R is a reduced-dose, highly effective active ingredient that exceeds the expected efficacy of standard metformin (R'), characterized by a significantly increased drug effect when the R-course dosage (QR) is ≤25%-50% of the R'-course dosage (QR'), including a significant improvement in drug efficacy indicators, manifested as a tumor inhibition rate or objective response rate in mouse experiments that is at least 25%-50% higher than that of R'.

[0363] In one embodiment, this application provides a combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, wherein the combination of R and Z (R / Z combination) is a reduced-dose, high-efficiency combination that exceeds the expected effect of the standard metformin drug (R') and Z combination (R' / Z combination). This is manifested in a significantly increased drug effect when the R-course dosage (QR) is ≤25%-50%, the R'-course dosage (QR') and the Z-course dosage are not increased, including a significantly improved drug efficacy index, manifested in a tumor inhibition rate or objective response rate in mouse experiments that is at least 10%, 20%, 30%, or 40% higher than that of the R' / Z combination.

[0364] In one embodiment, this application provides a further dose-reduction and synergistic combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, wherein the combination of R and Z (R / Z combination) is a further dose-reduction rapid-acting combination that exceeds the expected efficacy of R monotherapy and Z monotherapy using the HSA or Bliss method, characterized by a significant increase in drug effect when the dose of R for one course of treatment (QR) is not increased and the dose of Z for one course of treatment (QZ) is reduced by at least 20%, including an improvement in drug efficacy indicators, manifested as a tumor inhibition rate or objective response rate in mouse trials that is at least 10%-20% higher than the highest of the monotherapy.

[0365] In one embodiment, the dose-reduced high-efficiency regimen described in this application is preferably applicable to tumors requiring the strongest possible dose-response and the highest possible efficacy, including, for example, highly aggressive tumors, advanced tumors, rapidly progressing tumors, drug-resistant tumors, tumors with unfavorable microenvironments, and tumors that have discontinued treatment. If the standard metformin regimen is chosen instead of the dose-reduced high-efficiency regimen of this application, there is a significant clinical risk of increased drug dosage due to misuse of the regimen, resulting in low efficacy and missed treatment opportunities.

[0366] 3) Long-lasting effect

[0367] In this application, the term "long-acting" refers to the ability to sustainably suppress a target disease or symptom, or to provide sustained benefit to the patient. The anticipated long-acting nature of a regimen can be long-acting, non-long-acting, etc., and is highly uncertain. Tumor growth is often difficult to control, and the duration of drug efficacy (e.g., anti-tumor drugs) is mostly short; the probability of long-acting drugs is far lower than that of non-long-acting drugs. This is a crucial objective of regimen research, as well as a significant technical challenge and essential characteristic. If a new regimen exceeds the anticipated long-acting nature of an older, non-long-acting regimen, it constitutes long-acting, which not only represents unexpected synergy but also provides important evidence of unique innovation in the treatment mechanism or drug design.

[0368] In this application, taking the mouse experiment of the embodiment as an example, the long-term effect evaluation is as follows.

[0369] (1) Calculate long-term effectiveness indicators

[0370] In this application, the term "long-term efficacy indicator" refers to progression-free time or long-term survival (hereinafter referred to as survival rate). Progression-free time is the number of days from the first day of administration to the day on which progression is observed, wherein the progression is evaluated by relative tumor volume (RTV), and progression is defined as an RTV that increases continuously until the endpoint when the RTV is >150%. Survival rate is the ratio of the number of survivors observed in the study group at the time when most deaths occur in the negative control group (e.g., day 30) to the number of enrolled patients, wherein, in accordance with this convention, tumor volumes >2000 mm3 are counted as deaths.

[0371] (2). Relative evaluation

[0372] Monotherapy comparison: In the prior art, R and R' are considered pharmaceutically equivalent and are expected to have the same long-lasting effect. The evaluation criterion is: when the actual progression-free time or survival rate of R monotherapy is at least 20%-40% higher than that of R' monotherapy, its long-lasting effect exceeds the expectations of the prior art and is considered long-lasting.

[0373] Combined Comparison: As mentioned above, the long-term efficacy of R / Z is consistent with that of R' / Z based on existing technology. The evaluation criterion is: if the actual time without progress or survival rate of R / Z is at least 20%-40% higher than that of R' / Z, then its long-term efficacy exceeds the expectations of existing technology and is considered long-term efficacy.

[0374] (B) Further comparison

[0375] In further comparisons, R / Z was determined to be the highest in terms of the combined long-acting expectation of single-drug R and Z (HSA method). The evaluation criterion was: if the actual progression-free survival or survival rate of R / Z was at least 10%-20% higher than the highest of R and Z single-drugs, then its long-acting effect further exceeded expectations, thus qualifying as further long-acting.

[0376] In this application, the terms "further long-term" (program), "long-term" (program), and "non-long-term" (program) refer to (programs) that meet the above evaluation criteria for further long-term, long-term, and non-long-term, respectively.

[0377] In one embodiment, the differential structure-activity relationship of the present application can be simply expressed as f: (α, β, γ) → (X, Y), wherein the differential indication (X, Y) is the range (X) to which the long-acting or further long-acting (Y) of R or R / Z is applicable, while the non-long-acting (X) of R' or R' / Z is not applicable: patients with local lesions who require prolonged therapeutic effect; the differential pharmacology (f) mainly includes the long-acting or further long-acting transient local effects of R that are different from those of R'; the differential component set (α, β, γ) is the selection and range of long-acting or further long-acting components defined by (X, Y) and f.

[0378] In one embodiment, this application provides a formulation, method, kit, or use for treating local lesions, wherein R is a reduced-dose long-acting active ingredient that exceeds the expected efficacy of standard metformin (R'), characterized by a significant increase in drug effect when the dosage of R for one course of treatment (QR) is ≤25%-50% of the dosage of R' for one course of treatment (QR'), including a significant improvement in drug long-acting indicators, manifested as a progression-free time or survival rate in mouse experiments that is at least 20%-40% higher than that of R'.

[0379] In one embodiment, this application provides a combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, wherein the combination of R and Z (R / Z combination) is a reduced-dose, high-efficiency combination that exceeds the expected effect of the standard metformin drug (R') and Z combination (R' / Z combination), characterized by a significantly increased drug effect when the R-course dosage (QR) is ≤25%-50% and the R'-course dosage (QR') and Z-course dosage are not increased, including a significantly improved drug long-acting index, manifested as a progression-free time or survival rate in mouse experiments that is at least 20%-40% higher than that of the R' / Z combination.

[0380] In one embodiment, this application provides a further dose-reduction and synergistic combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, wherein the combination of R and Z (R / Z combination) is a further dose-reduction rapid-acting combination that exceeds the expected efficacy of R monotherapy and Z monotherapy using the HSA or Bliss method, characterized by a significant increase in drug effect when the dose of R for one course of treatment (QR) is not increased and the dose of Z for one course of treatment (QZ) is reduced by at least 20%, including an improvement in drug long-acting indicators, manifested as a progression-free time or survival rate in mouse studies that is at least 10%-20% higher than the highest of the monotherapy.

[0381] In one embodiment, the reduced-dose long-acting regimen of this application is preferably applicable to tumors requiring the strongest possible dose-response and the longest possible duration of action, including, for example, highly aggressive tumors, advanced tumors, rapidly progressing tumors, drug-resistant tumors, tumors with unfavorable microenvironments, and tumors that have discontinued treatment. If the standard metformin regimen is chosen instead of the potent long-acting regimen of this application, there may be numerous risks in clinical practice, such as increased drug dosage due to misuse of the regimen, resulting in low efficacy, shorter progression-free survival, and thus missed treatment opportunities.

[0382] 4) Local efficacy-toxicity ratio

[0383] In this application, the term "local efficacy-to-toxicity ratio" (or simply efficacy-to-toxicity ratio) refers to the balance between the therapeutic effect (local efficacy) and the toxicity (local toxicity) of a drug when administered locally. It is generally believed that the local efficacy required to produce the desired effect (e.g., anhydrous ethanol) is consistent with its local toxicity. Improving the efficacy-to-toxicity ratio and breaking this expected consistency is a key objective of protocol studies, as well as a significant technical challenge and essential characteristic. If a new protocol exceeds the expected efficacy-to-toxicity ratio of an older protocol, it not only demonstrates unexpected synergy but also provides crucial evidence of unique innovation in the treatment mechanism or drug design.

[0384] In this application, taking the mouse experiment of the embodiment as an example, the efficacy-toxicity ratio study was mainly carried out under the condition of Z=C, and the evaluation is as follows.

[0385] (1). Effect-toxicity ratio index

[0386] In this application, the term "efficacy-to-toxicity ratio" refers to the ratio (X / Y) of the desired therapeutic effect (X, selected from efficacy, such as a tumor inhibition rate of >40%) after the local application of an equal amount of drug to the local tissue irritation response (Y, local irritation score). When local administration of a drug produces neither significant therapeutic effect (e.g., tumor inhibition rate <20%) nor significant local irritation (e.g., score <1.0), its efficacy-to-toxicity ratio is considered to be close to zero in this application.

[0387] (2). Relative evaluation

[0388] (A) Standard Comparison

[0389] The evaluation criteria are as follows: when the amount of at least one component in R / C is reduced by 50% compared with the amount of the corresponding component in R' / C, and its efficacy-toxicity ratio (such as X / Y) is increased by at least 0.5 times, then the efficacy-toxicity ratio of R / C is considered to exceed the expectations of the prior art (R / C and R' / C are pharmaceutical equivalents) and is considered to be "high efficacy-toxicity ratio".

[0390] (B) Further comparison

[0391] In further comparisons, the expected efficacy-toxicity ratio (q2) was calculated using the HSA method for single-drug R and C. The evaluation criterion was: when the ratio of the actual efficacy-toxicity ratio (q1) to the expected ratio (q2) of R / C (q=q1 / q2) > 1, it was considered a further improvement in efficacy-toxicity ratio beyond expectations.

[0392] In one embodiment, the differential structure-activity relationship of the present application can be simply expressed as f: (α, β, γ) → (X, Y), wherein the differential indication (X, Y) is the range (X) to which the high-efficiency-toxicity ratio or further high-efficiency-toxicity ratio (Y) of the R or R / C is applicable, while the R' or R' / C is not applicable due to ineffectiveness: patients with local lesions who are intolerant to local irritation; the differential pharmacology (f) mainly includes the transient local effects of the R that are different from the R' in terms of high-efficiency-toxicity ratio or further high-efficiency-toxicity ratio; the differential component set (α, β, γ) is the selection and range of high-efficiency-toxicity ratio or further high-efficiency-toxicity ratio components defined by (X, Y) and f.

[0393] In one embodiment, this application provides a combination, pharmaceutical composition, formulation, method, kit, or use for treating local lesions, wherein the combination of R and C is a high efficacy-toxicity ratio combination that exceeds the expected effect of the combination of R' and C, characterized in that the actual efficacy-toxicity ratio (q1) of R / C exceeds the expected ratio (q2) based on the equivalence of R and R'.

[0394] In one embodiment, this application provides a further dose-reduction synergistic combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, wherein the combination of R and C is a further synergistic combination beyond what is expected by the HSA method based on R and C monotherapy, characterized by the actual efficacy-toxicity ratio (q1) of the combination exceeding the expected ratio (q2) based on R and C monotherapy.

[0395] In one embodiment, the high-efficiency-to-toxicity ratio regimen or even a more high-efficiency-to-toxicity ratio regimen of this application is preferably suitable for patients, tumors, or tumor sites requiring treatment with a regimen that requires the highest possible efficacy-to-toxicity ratio, including, for example, highly aggressive tumors, advanced tumors, rapidly worsening tumors, refractory tumors, or sites in which said tumors are prone to drug leakage. If the regimen of this application is not selected in favor of the aforementioned control regimen, numerous risks of damage to normal tissues may arise clinically due to misuse of the regimen (choosing the opposite when a high efficacy-to-toxicity ratio should be preferred).

[0396] 4. Heterogeneous adaptation

[0397] In this application, the term "heterogeneous adaptability" refers to the ability of a regimen to exhibit convergent therapeutic effects (e.g., rapid-acting, long-acting, highly effective) in heterogeneous regions of a target disease or symptom (e.g., different tissues or lesions). The term "adjustability" refers to the ability of a regimen to adjust its therapeutic effect or administration parameters according to dynamic changes in the disease state (including the aforementioned heterogeneity) or changes in external conditions. The expected heterogeneity adaptability or adjustability of a regimen can be highly adaptive / highly adjustable, generally adaptive / adjustable, maladaptive / unadjustable, etc., and is highly uncertain.

[0398] One of the fundamental characteristics of malignant tumors is their high heterogeneity, and standard antitumor regimens, including metformin, often suffer from heterogeneity adaptation issues. Tumor-administered regimens (such as anhydrous ethanol regimens) also typically exhibit heterogeneity adaptation problems, making it difficult to tailor medication to individual tumor types. Heterogeneity includes tissue heterogeneity, spatial heterogeneity, and safety heterogeneity. In short, improving heterogeneity adaptation or adjustability is a crucial objective, a significant technical challenge, and an essential characteristic of antitumor drug technology research. If a new regimen exceeds the expected adjustability of the old one, it not only provides for unexpected synergistic effects but also allows for adjustments and optimization under different conditions to adapt to the flexibility and diversity required for individualized clinical treatment. Furthermore, it provides important evidence of unique innovation in treatment mechanisms or drug design.

[0399] In this application, taking the mouse experiment of the embodiment as an example, the evaluation of heterogeneous adaptability / adjustability is as follows.

[0400] 1) Calculate the heterogeneity / adjustability index

[0401] In this application, the terms "heterogeneous adaptability index" or "adjustability index" refer to the differences in therapeutic effects of a regimen in different tissues or lesions (different tumors, different lesions, etc.), or the adjustability coefficients of local application parameters that achieve convergent target therapeutic effects in the same tissue or lesion. These coefficients are all ratios of the highest value in the adjustable range to its threshold, including the local application concentration adjustability coefficient (referred to as concentration adjustability coefficient, e.g., highest value of cR / threshold), the local application target volume adjustability coefficient (referred to as target volume adjustability coefficient, highest value of vtarget / threshold), the local application volume ratio adjustability coefficient (referred to as volume ratio adjustability coefficient, e.g., highest value of vR / vtarget / threshold), the local application single dose adjustability coefficient (referred to as dose adjustability coefficient, e.g., highest value of cR×vR / threshold), the local application point density adjustability coefficient (referred to as application point density adjustability coefficient, e.g., highest value of ni / vtarget / threshold), etc.

[0402] 2) Relative evaluation

[0403] (1). Standard Comparison

[0404] Monotherapy comparison: The evaluation criteria are as follows: when R' monotherapy is effective against different tissues or lesions, or when the actual value of any of the above-mentioned adjustable coefficients of R monotherapy is at least 100% or 300% higher than that of R' monotherapy, then its heterogeneity adaptation or adjustability exceeds the expectations of the prior art (R and R' are pharmaceutical equivalents) and is highly adjustable. According to the teachings of the prior art, the therapeutic effects of R' monotherapy on different tissues or lesions (different tumors, different lesions, etc.) vary greatly (only difficult for a very few tumors), and even for the very few tumors for which it is effective, the efficacy of R' is dose-dependent rather than locally applied parameter-dependent, and its local adjustability coefficient is not high.

[0405] Combination comparison: The evaluation criteria are as follows: when R / Z is effective for different tissues or lesions, or when the actual value of any of the above-mentioned adjustable coefficients of R / Z is at least 100% or 300% higher than that of R' / Z, then its heterogeneity adaptation or adjustability exceeds the expectations of the prior art (R / Z and R' / Z are pharmaceutical equivalents) and is highly adjustable; otherwise, it is not highly adjustable.

[0406] (2). Further comparison

[0407] In further comparisons, the R / Z ratio is determined by the highest shared adjustability of the expected adjustability of individual drugs R and Z (HSA method). The evaluation criterion is: if the actual value of any of the above adjustability coefficients of R / Z is at least 20% higher than the highest value of R and Z among individual drugs, then its adjustability exceeds expectations and is considered to be further highly adjustable.

[0408] In this application, the terms "further highly adjustable" (scheme), "highly adjustable" (scheme), and "non-highly adjustable" (scheme) refer to (schemes) that meet the above evaluation criteria for further highly adjustable, highly adjustable, and non-highly adjustable, respectively, and they respectively provide drug effects of further highly heterogeneous equivalence, highly heterogeneous equivalence, and non-highly heterogeneous equivalence.

[0409] In one embodiment, the differential structure-activity relationship of the present application can be simply expressed as f: (α, β, γ) → (X, Y), wherein the differential indication (X, Y) is the range (X) to which the highly adjustable or further highly adjustable R or R / Z is applicable, while the non-highly adjustable R' or R' / Z is not applicable: patients with local lesions who require heterogeneous adaptability and adjustability of the drug to optimize the treatment regimen; the differential pharmacology (f) mainly includes the highly adjustable or further highly adjustable transient local effects of R that differ from those of R'; the differential component set (α, β, γ) is the selection and range of highly adjustable or further highly adjustable components defined by (X, Y) and f.

[0410] In one embodiment, this application provides a formulation, method, kit, or use for treating local lesions, wherein R is a dose-reduced synergistic active ingredient that exceeds the expected effect of standard metformin drug (R), characterized by a significant increase in drug effect when the dose of R for one course of treatment (QR) is ≤25%-50% of the dose of R' for one course of treatment (QR'), including a significant increase in at least one drug adjustability index, characterized by an increase of at least 100% or more in at least one of the following adjustability coefficients in mouse experiments: concentration adjustability coefficient, target volume adjustability coefficient, volume ratio adjustability coefficient, single dose adjustability coefficient, and application point density adjustability coefficient.

[0411] In one embodiment, this application provides a combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, wherein the combination of R and Z (R / Z combination) is a dose-reduction synergistic combination that exceeds the expected effect of the standard metformin drug (R') and Z combination (R' / Z combination), characterized by a significant increase in drug effect when the R-course dosage (QR) is ≤25%-50%, the R'-course dosage (QR') and the Z-course dosage are not increased, including a significant increase in at least one drug adjustability index, characterized by an increase of more than 100% in at least one of the following adjustability coefficients in mouse experiments: concentration adjustability coefficient, target volume adjustability coefficient, volume ratio adjustability coefficient, dose adjustability coefficient, and application point density adjustability coefficient.

[0412] In one embodiment, this application provides a further dose-reduction synergistic combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, wherein the combination of R and Z (R / Z combination) is a further dose-reduction synergistic combination that exceeds the expected efficacy of R monotherapy and Z monotherapy using the HSA or Bliss method, characterized by a significant increase in drug effect when the dose of R for one course of treatment (QR) is not increased and the dose of Z for one course of treatment (QZ) is reduced by at least 20%, including a significant increase in at least one drug adjustability index, characterized by at least 10%-20% higher than the highest of the monotherapy in mouse studies of at least one of the following adjustability coefficients: concentration adjustability coefficient, target volume adjustability coefficient, volume ratio adjustability coefficient, dose adjustability coefficient, and application point density adjustability coefficient.

[0413] In one embodiment, the local application concentration adjustment factor (maximum / threshold value of cR) of said R is >10, 50, 100, or 150, which allows said R to provide highly variable concentrations to different regions while still maintaining its said activity, including, for example, individualized administration to heterogeneous tumors.

[0414] In one embodiment, the local application target volume adjustment factor (maximum value / threshold of target v) of said R is >2, 10, 50, or 100, which allows said R to select different targets v for different treatment situations, including, for example, heterogeneous tumors, targeted drug administration.

[0415] In one embodiment, the local application volume ratio of said R is adjustable (vR / vtarget maximum / threshold) is >5, 10, or 100, which allows said R to provide highly variable volume ratios to different regions while still maintaining its activity stability, including, for example, heterogeneous tumors, targeted drug delivery.

[0416] In one implementation, the local application dose adjustment factor (maximum value of cRi×vRi / threshold) of said R is >2, 6, or 10, which allows said R to be selected at different single doses for different treatment situations, including, for example, individualized treatment for heterogeneous tumors; and

[0417] In one embodiment, the local application point density adjustable coefficient (nii / v target maximum / threshold) of said R is >3, 10, 50, or 100, which allows said R to provide highly variable density to different regions while still maintaining its said activity stability, including, for example, individualized drug delivery to heterogeneous tumors.

[0418] In this application, the terms "micro-volume" or "micro-amount" (scheme) refer to a scheme in which the vapplication / vtarget ratio is 0.02-0.056 or 0.02-0.10 in a locally applied volume ratio height-adjustable (scheme); otherwise, it is a "constant volume" (scheme). The term "high density" (scheme) refers to a scheme in which the ni / vtarget ratio is ≥1 or 2 points per cm3 in a locally applied point density height-adjustable (scheme); otherwise, it is a "constant density" (scheme).

[0419] In one embodiment, this application provides a formulation, method, kit, or use for treating solid tumors, which is a highly adjustable local application volume scheme, including microvolume application with a vapplication / vtarget ratio of 0.02-0.056 or 0.02-0.10. It is suitable for differentiated intratumoral administration or administration based on tumor characteristics, including administration based on shape (shape heterogeneity), administration based on tissue (tissue heterogeneity), or administration based on safety risk (safety risk heterogeneity).

[0420] In one embodiment, this application provides a combination, pharmaceutical composition, formulation, method, kit, or use for treating solid tumors, which is a highly adjustable local application point density scheme, including high-density application of nii / v target at ≥1 point per cm3. It is suitable for differentiated intratumoral administration or administration based on tumor characteristics, including administration based on shape (shape heterogeneity), administration based on tissue (tissue heterogeneity), or administration based on safety risk (safety risk heterogeneity).

[0421] In one implementation, for larger lesions (e.g., long diameter > 2 cm), especially large lesions (e.g., long diameter > 3 cm), the height adjustability includes multi-point application due to tissue heterogeneity, multi-point application due to morphological heterogeneity, and / or multi-point application due to safety risk heterogeneity, wherein the multi-point refers to the number of application points being > 3, > 5, or 10-200, and at least one of them being a micro-volume application.

[0422] In one embodiment, the height adjustability includes applying a low concentration (R application concentration ≤ 3%) or / and a microvolume (R application volume ≤ 250 μL) near the edge of the lesion.

[0423] In one embodiment, the height adjustability includes applying a high concentration (R application concentration ≥ 10%) or / and a large volume (R application volume ≥ 1000 μL) near the center of the lesion.

[0424] In one embodiment, the dose-reduction and efficacy-enhancing regimen of this application is preferably applicable to heterogeneous tumors requiring the strongest possible dose-response and the highest possible adjustability, including, for example, highly aggressive tumors, advanced tumors, rapidly progressing tumors, drug-resistant tumors, tumors with unfavorable microenvironments, and tumors that have discontinued treatment. If the standard metformin regimen is chosen instead of the potent, long-acting regimen of this application, there is a significant risk in clinical practice of increased drug dosage due to misuse of the regimen, resulting in low efficacy and limited adjustability for individual tumor types, thereby missing crucial treatment opportunities.

[0425] Other terms used herein are intended to be defined by their meanings as known in the art.

[0426] In this application, the term "lesion" refers to a localized abnormality in the structure or function of the body, typically including diseased tissue, which in turn contains cells. A lesion is a localized manifestation of a disease.

[0427] In this application, the term "intervention" is distinguished from standard injection and refers to the administration (drug delivery) of a therapeutic substance to a lesion site through a specific application device (such as a puncture needle, injection needle, catheter or other) or other mediator (such as a natural body orifice or minimally invasive incision), including vascular intervention (such as transvascular perfusion) and non-vascular intervention (such as percutaneous injection).

[0428] The term "standard injection" refers to a method of administration that does not exclude, or even primarily excludes, or prefers, absorption injection; the term "absorption injection" refers to an injection method that allows the drug to be absorbed into the bloodstream after entering the body, including, for example, intravenous injection, intramuscular injection, and intraperitoneal injection.

[0429] The term "standard injection safety" refers to one of the quality standards of standard injection, namely, that it does not produce local side effects (such as local irritation, local tissue damage, etc.) during standard injection. Therefore, it must have properties such as isotonicity, isotonicity, and isoacidity (pH 4.5-9.0).

[0430] In this application, the term "dedicated" means that it cannot be used, or is even strictly prohibited, for any purpose other than the specified purpose. For example, "dedicated for intervention" means that it is limited to interventional use and excludes use for absorption injection.

[0431] In this application, the term "medicinal solution" refers to an aqueous solution of a drug that can be injected into the human body to produce a therapeutic effect, containing the active ingredient and its aqueous solvent.

[0432] The term "composition" (or structure, or simply "structure") refers to the constituent elements necessary to provide a specific function ("effect") that cannot be selected according to known knowledge, including, for example, the active ingredient, its excipients, and the concentration at which they are introduced must be limited.

[0433] The term "concentration" refers to the weight percentage concentration (W component / W solution, %) or molar concentration (W component / M component / 1 liter) of a specified component in a solution.

[0434] The term "interventional concentration" refers to the concentration of a specified component in the drug solution at the time of intervention (e.g., at the injection site or infusion tube outlet).

[0435] In this application, the term "prior art" (or "standard" (e.g., drug, injection, dosage form, method, use, or reaction) refers to the scope of existing predictions (e.g., drug, injection, dosage form, method, use, or reaction).

[0436] In this application, the term "transient active ingredient" is distinguished from "persistent active ingredient," the former referring to active ingredients that provide transient and persistent effects, respectively. Examples of the former include chemical ablation agents, while examples of the latter include standard antitumor drugs (including antitumor drugs). The term "transient effect" is distinguished from "persistent effect," the former referring to the effect provided by an active ingredient that is maintained at a specific concentration for a short time and the latter for a relatively long period of time. The required duration of action is relatively short and relatively long, respectively. They are independent of pharmacokinetics (e.g., half-life) and closely related to pharmacokinetics (e.g., half-life), respectively. They are the effects that appear faster and the effects that appear later, respectively. Examples of the former include chemical ablation agents, while examples of the latter include standard effects (including cytotoxic effects).

[0437] Within the scope of this invention, the term "tumor" refers to a pathological disease characterized by a lumpy mass, which can be a tumor of any pathology (malignant and non-malignant) and at any stage, including, for example, the following groups classified according to tumor cell type: epithelial cell tumors, sarcomas, lymphomas, germ cell tumors, germ cell tumors; and tumors named according to the organ or tissue in which the tumor cell concentration area is located, including, for example, tumors named according to the following organs or tissues: skin, bone, muscle, breast, kidney, liver, lung, gallbladder, pancreas, brain, esophagus, bladder, large intestine, small intestine, spleen, stomach, prostate, testicle, ovary, or uterus.

[0438] Specifically, the malignant tumors include, for example, breast cancer, pancreatic cancer, thyroid cancer, nasopharyngeal cancer, prostate cancer, liver cancer, lung cancer, intestinal cancer, oral cancer, esophageal cancer, stomach cancer, laryngeal cancer, testicular cancer, vaginal cancer, uterine cancer, ovarian cancer, sarcoma, etc.

[0439] 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, stomach tumors, nasopharyngeal tumors, laryngeal tumors, testicular tumors, vaginal tumors, uterine tumors, fallopian tube tumors, ovarian tumors, etc.

[0440] The drug described in this application is a therapeutic drug that serves as the primary treatment for solid tumors. It can also be used in combination with other interventional therapies, systemic chemotherapy, immunotherapy, photodynamic therapy, sonodynamic therapy, surgical intervention, or combinations thereof to further enhance efficacy.

[0441] According to one aspect of the present invention, a metformin(R) monotherapy product (R monotherapy regimen 1) is provided, specifically as a formulation (hereinafter referred to as "the R monotherapy formulation of the present application") comprising N,N-dimethylbiguanide and optionally a solvent, wherein the formulation does not contain an acidic pH adjuster required to meet the safety requirements for cell culture or standard injection, wherein the concentration of N,N-dimethylbiguanide contained in the formulation is ≥ its intervention or administration concentration, wherein the intervention or administration concentration (w / w) is 0.5% ≤ C ≤ 51%, preferably 1%-50%, more preferably 3%-50%, 5%-50%, or 5%-30%.

[0442] The present invention provides a single-drug R product (Single-Drug R Recipe 2) specifically for interventional treatment of lesions, comprising metformin solution (hereinafter referred to as "the R single-drug solution of this application"), which may be an aqueous solution of N,N-dimethylbiguanide, wherein the aqueous solution does not contain an acidic pH adjuster required to meet the safety requirements for cell culture or standard injection, and the concentration of the N,N-dimethylbiguanide solution {〔W metformin / (W metformin + W solvent)〕%} is ≥ its interventional concentration, wherein the interventional concentration is 0.5% ≤ C ≤ N,N-dimethylbiguanide maximum solubility concentration (or saturation concentration).

[0443] This invention provides a metformin monotherapy drug (R monotherapy regimen 3) specifically for interventional lesion treatment (hereinafter referred to as "the R monotherapy drug of this application"). It is an aqueous solution of N,N-dimethylbiguanide, wherein the aqueous solution does not contain an acidic pH adjuster required for safety in cell culture or standard injection, and the concentration of the N,N-dimethylbiguanide monotherapy solution {〔W metformin / (W metformin + W solvent)〕%} is ≥ its interventional concentration, wherein the interventional concentration is 0.5% ≤ C ≤ the maximum solubility concentration of N,N-dimethylbiguanide. In fact, the R monotherapy drug of this application is a drug whose interventional form is the R monotherapy drug solution of this application.

[0444] This invention provides a metformin R monotherapy formulation (R monotherapy regimen 4) specifically for interventional lesion treatment (hereinafter referred to as "the R monotherapy formulation of this application"). It is an aqueous solution of N,N-dimethylbiguanide, wherein the aqueous solution does not contain an acidic pH adjuster required for safety in cell culture or standard injection, and the concentration of the N,N-dimethylbiguanide R monotherapy solution {〔W metformin / (W metformin + W solvent)〕%} is ≥ its interventional concentration, wherein the interventional concentration is 0.5% ≤ C ≤ N,N-dimethylbiguanide's maximum solubility concentration (saturation concentration). In fact, the R monotherapy formulation of this application is a convenient formulation for interventional treatment of lesions using the R drug of this application.

[0445] In the preparations, solutions, drugs, and / or drug dosage forms described above, the intervention concentration (w / w) is 0.5% ≤ C ≤ 51%, preferably 1%-50%, more preferably 3%-50%, 3%-40%, 5%-50%, 5%-35%, or 5%-30%.

[0446] The preparations, solutions, drugs, and drug formulations described above are used for interventional treatment of lesions such as tumors or nodules in subjects.

[0447] In the formulation described above, the formulation contains a solvent, preferably water, more preferably water for injection.

[0448] The formulations, solutions, drugs, or drug dosage forms described above do not include salts of N,N-dimethylbiguanide.

[0449] In the formulations, solutions, drugs, and / or drug dosage forms described above, the administration or intervention concentration (w / w) of metformin is ≤51%, ≤50%, ≤40%, ≤30%, ≤25%, ≤20%, ≤10%, ≤5%, or ≤2%, such as 1%, 0.75%, or 0.5%.

[0450] In the formulations, solutions, drugs, and / or drug dosage forms described above, the administration or intervention concentration (w / w) of metformin (R) is 0.2%-51%, 0.5-50%, 1%-40%, 5%-50%, 5%-40%, 5%-30%, 5%-25%, 5%-20%, 1%-30%, 1%-25%, 1%-20%, 0.5%-10%, 1%-5%, or ≤2%, such as 1%, 0.75%, or 0.5%.

[0451] In the preparations, solutions, drugs, and / or drug dosage forms described above, the acidic pH adjuster includes or is selected from hydrochloric acid, sulfuric acid, acetic acid, citric acid, phosphoric acid, malic acid, tartaric acid, lactic acid, and carbonic acid.

[0452] In this application, the formulation, liquid, drug, or dosage form of this application is referred to as "the R monotherapy drug" (R monotherapy regimen).

[0453] The single-drug product in this application is an interventional-specific product, not a general injectable, and is even strictly prohibited from use in standard injections (e.g., it must be stated in the instructions for use).

[0454] According to one embodiment, the R-single-drug solution in the R-single-drug pharmaceutical product of this application is a solution used to provide the transient effect of metformin.

[0455] According to one embodiment, the single-drug formulation of this application is a low-frequency, rapid-acting interventional formulation. Existing metformin formulations are primarily for frequent, long-acting therapy, and metformin's current anti-lesion, such as anti-cancer responses, cannot provide low-frequency rapid-acting effects; at most, they can only provide high-frequency long-acting effects. Conversely, the low-frequency, rapid-acting interventional formulation of this application is strictly prohibited from high-frequency injection (e.g., it must be stated in the instructions for use) and is not a very frequent, long-acting injection formulation; the former must provide the low-frequency rapid-acting effect that the latter cannot anticipate. The difference between the two is further expanded based on the aforementioned structure-activity characteristics: the latter must provide high-frequency long-acting effects, including the benefits and safety of high-frequency administration and efficacy appearing only after a long period, which must be implemented by a structural design that further meets the safety requirements of high-frequency, long-cycle standard injections (e.g., minimizing local irritation); the former must provide low-frequency rapid-acting effects, which can only be implemented by a structural design that deviates further from the standard injection safety requirements (1.5% ≤ CA < 10%).

[0456] According to one embodiment, the low-frequency, fast-acting interventional formulation of this application is further applicable to patients who are not suitable for high-frequency intervention and are not suitable for conventional treatment, based on the indications of the single-drug formulation of this application mentioned above. For example, patients with poor interventional compliance who urgently need therapeutic effects (not suitable for patients who still have no obvious therapeutic effect after a long period of time, such as those whose condition is progressing rapidly or who are receiving palliative treatment).

[0457] According to one embodiment, the metformin powder for injection of this application R is a special powder for interventional use, comprising sterile powder and solvent required for preparing the drug solution of this application, wherein the sterile powder comprises N,N-dimethylbiguanide and is contained in a container filled with inert gas; the solvent comprises a pharmaceutically acceptable carrier, such as a solvent or medium, wherein the solvent is a reagent for dissolving N,N-dimethylbiguanide, such as water for injection, and has a volume sufficient to dissolve the N,N-dimethylbiguanide to the concentration stated in the single-drug solution of this application R. The metformin powder for injection of this invention is strictly prohibited from use as a standard powder for injection.

[0458] According to another aspect of the present invention, the present invention provides the use of N,N-dimethylbiguanide and its monotherapy solution (R monotherapy regimen 2), specifically:

[0459] This invention provides the use of the drug solution of this application as a transient agent in the preparation of the drug or dosage form of this application (hereinafter referred to as "the use of the drug solution of this application R"), wherein the drug solution is used to provide the N,N-dimethylbiguanide on diseased tissue and its contained cells.

[0460] This invention provides the use of N,N-dimethylbiguanide as a transient active ingredient in the preparation of interventional solutions, drugs, or dosage forms for lesions (hereinafter referred to as "metformin monotherapy use in this application"), wherein the N,N-dimethylbiguanide is used to provide a transient effect on lesion tissue and its contained cells.

[0461] This invention provides a method for treating cells within a lesion (hereinafter referred to as "the single-drug cell treatment method of this application"), comprising the following steps: intervening in the lesion with a special N,N-dimethylbiguanide solution or contacting the lesion with the N,N-dimethylbiguanide of this invention, wherein the N,N-dimethylbiguanide is used as a transient active ingredient to provide a transient effect on the lesion tissue and its contained cells.

[0462] This invention provides a method for treating lesions (hereinafter referred to as "the single-drug lesion treatment method of this application"), comprising the following steps: intervening in the lesion with a special N,N-dimethylbiguanide interventional solution, or contacting the lesion with the N,N-dimethylbiguanide of this invention, or applying N,N-dimethylbiguanide to the lesion, wherein the N,N-dimethylbiguanide serves as a transient active ingredient to provide a transient effect on the lesion tissue and its contained cells. In fact, the lesion treatment method of this application includes the cell treatment method of this application.

[0463] This invention provides a method for treating a lesion (hereinafter referred to as "the single-drug disease treatment method 1 of this application"), comprising the following steps: intervening in the lesion with a special N,N-dimethylbiguanide interventional solution, or contacting the lesion with the N,N-dimethylbiguanide of this invention, or applying N,N-dimethylbiguanide to the lesion, wherein the N,N-dimethylbiguanide serves as a transient active ingredient to provide a transient effect on the lesion tissue and its contained cells. In fact, the single-drug disease treatment method of this application R includes the single-drug lesion treatment method of this application R.

[0464] This invention provides a method for treating lesions such as tumors or nodules in a subject (hereinafter referred to as "the single-drug disease treatment method of this application R"). This method includes intervening in or administering the single-drug medicine or preparation of this application R as described above to the subject.

[0465] In some embodiments, the intervention or application can be systemic or local. Preferably, the application or intervention can include intratumoral application or intervention, preferably local intratumoral intervention, preferably local injection application or intervention into the tumor or tumor microenvironment.

[0466] In some implementations, injection administration may include infusion, liquid particle injection, or implantation.

[0467] In some preferred embodiments, the formulations described above are suitable for micro-volume administration or interventional micro-volume administration to reduce (Y) lesions (X), wherein the ratio of the administration volume to the target volume (V administration / V target) is approximately 0.02-0.056, 0.02-0.10, or 0.02-0.34.

[0468] In some embodiments, the above-described contact, intervention, or application includes multi-point contact, intervention, or application, preferably local multi-point contact or intervention.

[0469] The aforementioned contact or application may include multiple (site) contact, application, or intervention. The multiple sites refer to the number of micro-volume intervention sites where the total target area efficacy evaluation is observed to be significantly higher than that of the standard metformin drug. Specifically, the multiple sites are, for example, multiple sites at the target area, such as ≥5, ≥10, ≥20, ≥30, ≥50 sites, such as at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15 or more sites, such as 10 to 20, 10-30, 10-40, 10-50, 20-30, 20-40, 20-50, 25-50, 30-50 or more sites. Preferably, the contact points, application sites, or intervention sites are evenly distributed, especially to areas where standard intervention cannot diffuse.

[0470] In some preferred embodiments, when the lesion is ≥3.5 cm3 or the maximum size is ≥2 cm, the number of sites is ≥5, such as 10, 15, 20, 25, 30, 35 or 50 sites.

[0471] In some preferred embodiments, the aforementioned contact, application, or intervention may include micro-volume multi-point contact, application, or intervention. Preferably, micro-volume multi-point injection, wherein the micro-volume for lesions larger than 3 cm includes amounts from 5 μL to 1000 μL, such as 5 μL-500 μL, 5 μL-250 μL, 5 μL-200 μL, 5 μL-100 μL, 5 μL-75 μL, 5 μL-50 μL, 5 μL-25 μL. The dosage ranges are L, 5μL-20μL, 10μL-200μL, 10μL-100μL, 10μL-75μL, 20μL-200μL, 20μL-100μL, 20μL-75μL, 30μL-200μL, 30μL-100μL, 40μL-200μL, 40μL-100μL, 50μL-200μL, 50μL-100μL, or 50μL-75μL, etc. Preferably, the dosage is applied through micro-volume multi-point contact or administration; more preferably, it is administered via micro-volume multi-point injection.

[0472] In some implementations, the above treatment can be sustained for at least one course of treatment. The duration of one course of treatment can be approximately 7 to 40 days, for example, approximately 9 to approximately 37 days, for example, approximately 15, 20, 25, 27, 28, 29, 30, 31, 32, 33, 34, or 35 days.

[0473] In one embodiment, during a course of treatment, contact with or application of the formulation of the present invention may be at a low frequency, such as about 1-6 times per course of treatment, preferably 1-5 times per course of treatment, 1-4 times per course of treatment, 1-3 times per course of treatment, or 1-2 times per course of treatment.

[0474] In this application, the use of the monotherapy drug solution of this application R or the use of metformin monotherapy of this application is referred to as the use of monotherapy drug of this application R; the cell treatment method of monotherapy drug of this application R, the lesion treatment method of this application R, or the disease treatment method of this application R is referred to as the treatment method of monotherapy drug of this application R; the use of monotherapy drug of this application R or the treatment method of monotherapy drug of this application R is referred to as the use (or method) of monotherapy drug of this application R; the transient effect of N,N-dimethylbiguanide on lesion tissue and its contained cells is referred to as the effect of monotherapy drug of this application R.

[0475] In the above-mentioned uses of this application R as a single drug, or the cell treatment method of this application R as a single drug, the lesion treatment method of this application R as a single drug, or the disease treatment method of this application R as a single drug, the lesion includes malignant cells and / or non-malignant cells.

[0476] In the above-described uses and methods of the present invention, the lesion is a lesion for which standard treatment is ineffective or has failed, including at least one of the following groups: tumors for which standard treatment is ineffective or has failed; non-malignant nodules.

[0477] In the above-described uses and methods of the present invention, the tumors that are unresponsive or ineffective to standard treatment include at least one selected from the group consisting of: chemotherapy-resistant tumors, tumors with a high stromal-to-stromal ratio, tumors that are unresponsive or ineffective to standard chemotherapy, and patients who do not respond well to chemotherapy.

[0478] In the above-described uses and methods of the present invention, the non-malignant nodule is a nodule comprising at least one of the following non-malignant cells: connective tissue cells, secretory gland cells, and epithelial cells.

[0479] In the above-described uses and methods of the present invention, the lesion comprises a tumor containing non-drug-resistant malignant tumor cells.

[0480] In the above-described uses and methods of the present invention, the malignant tumor cells are non-metformin cytotoxicity-specific tumor cells.

[0481] According to one embodiment, the drug solution described in the single-drug use (or method) of this application R is the single-drug drug solution of this application R.

[0482] According to one embodiment, the R single-drug solution of this application is the R single-drug solution described in the R single-drug use (or method) of this application.

[0483] According to one embodiment, the transient effects described in the single-drug use (or method) of this application R include transient tissue penetration and transient cell deformation.

[0484] According to one embodiment, the cell deformation includes effective cell deformation.

[0485] In this application, the term "cell deformation" refers to an observable change in cell morphology (e.g., volume), which includes reversible and irreversible deformation, wherein irreversible deformation can develop into necrosis; the term "effective cell deformation" refers to cell deformation that can effectively cause necrosis of the tissue composed of said cells.

[0486] According to one embodiment, in the above method of this application, the selection scheme for the dosage of the single drug solution of R is: drug solution volume based on lesion volume × transient effective interventional concentration.

[0487] According to one embodiment, in the above method of this application, the selection scheme for the concentration of the single drug solution R is: ≥ transient effective intervention concentration.

[0488] According to one embodiment, in the above method of this application, the selection scheme for the frequency of use of the R single drug solution is: the number of times required for transient treatment of the lesion.

[0489] The R single-drug solution application scheme in this application breaks through or even deviates from the existing R' method. The former and latter drug solutions are used in closed (interventional) and open (any injection method that can form drug-containing blood, preferably absorption injection), respectively. The drug solution dosage selection schemes are (drug solution volume based on lesion volume × transient effective interventional concentration) and (patient blood volume or weight or surface area × sustained effective blood drug concentration), respectively. The drug solution concentration selection schemes are ≥ transient effective interventional concentration and (for safety requirements) preferably a lower concentration, respectively. The drug solution usage frequency selection schemes are (several times required for transient treatment of lesions) and (tens to hundreds of times required for metformin half-life and sustained effective blood drug concentration), respectively.

[0490] The above-mentioned technical solutions in this application have produced technical effects that exceed the expectations of existing methods, such as rapid effect, single-drug effectiveness, single-dose or low-frequency effectiveness, and effectiveness against lesions that are ineffective or intolerant to existing metformin drugs, etc.

[0491] According to one embodiment, in the drug solution of the single drug or formulation of this application R or the use (or method) of this application, the concentration (CR) of N,N-dimethylbiguanide is 1.0% ≤ CR ≤ N,N-dimethylbiguanide maximum solubility concentration (or saturation concentration).

[0492] According to one embodiment, in the drug solution of the single drug product or single drug formulation or single drug use (or method) of this application, the concentration (CR) of N,N-dimethylbiguanide is 3.0% ≤ CR ≤ N,N-dimethylbiguanide maximum solubility concentration (or saturation concentration).

[0493] According to one embodiment, in the drug solution of the single drug product of this application R or the single drug use (or method) of this application R, the concentration (CR) of N,N-dimethylbiguanide is 5.0% ≤ CR ≤ N,N-dimethylbiguanide maximum solubility concentration (or saturation concentration).

[0494] According to one embodiment, the single-drug drug or the single-drug use (or method) of this application R is the fast-acting solution, drug, dosage form, use, or method of the N,N-dimethylbiguanide.

[0495] According to one embodiment, the single-drug pharmaceutical product or the single-drug use (or method) of this application R is a low-frequency administration solution, drug, dosage form, use, or method of the N,N-dimethylbiguanide described in this application.

[0496] According to one embodiment, the indications (Y, X) of the single-drug product or the single-drug use (or method) of this application R, referred to herein as the indications of this application, are suitable for use in treating (Y) lesions or lesion symptoms (X), wherein the lesions contain malignant cells and / or non-malignant cells.

[0497] According to one embodiment, in the indications of this application, X is the volume of the lesion, and Y is an effective or rapid reduction.

[0498] According to one embodiment, in the indications of this application, the lesion is a lesion that is effective with standard metformin treatment but not rapid-acting, low-frequency effective, or / and effective with micro-volume intervention.

[0499] According to one embodiment, in the indications of this application, the lesion is a lesion in which metformin can be used as an adjunct therapy but is ineffective when used alone.

[0500] According to one embodiment, in the indications of this application, the lesion is a lesion for which metformin standard treatment is ineffective.

[0501] According to one embodiment, in the indications of this application, the lesion is a lesion that is ineffective or fails to respond to standard treatment.

[0502] According to one embodiment, in the indications of this application, the lesion that is ineffective or fails to respond to standard chemotherapy includes or is selected from at least one of the following groups: tumors refractory to standard chemotherapy drugs, and non-malignant nodules. According to one embodiment, in the indications of this application, the refractory to standard chemotherapy drugs includes or is selected from at least one of the following groups: drug-resistant tumors, tumors with unfavorable microenvironments, and tumors that have been discontinued from chemotherapy.

[0503] According to one embodiment, in the indications of this application, the drug-resistant tumor is a tumor containing drug-resistant malignant tumor cells.

[0504] According to one embodiment, in the indications of this application, the drug-resistant tumor is a tumor containing primary drug-resistant cells.

[0505] According to one embodiment, in the indications of this application, the drug-resistant tumor is a tumor containing a drug-resistant microenvironment.

[0506] According to one embodiment, in the indications of this application, the tumor containing a drug-resistant microenvironment is a tumor containing an unfavorable microenvironment.

[0507] According to one embodiment, in the indications of this application, the tumor containing an unfavorable microenvironment is a tumor in which the ratio of non-malignant cells to malignant cells is ≥43%.

[0508] According to one embodiment, in the indications of this application, the tumor that is ineffective or fails after chemotherapy is a tumor that is ineffective or fails after treatment with recognized first-line or second-line chemotherapy drugs.

[0509] According to one embodiment, in the indications of this application, the non-malignant nodule includes or is selected from at least one of the following non-malignant cell groups: connective tissue cells, secretory gland cells, and epithelial cells.

[0510] According to one embodiment, in the indications of this application, the connective tissue cells include or are selected from at least one of the following: fibroblasts, adipocytes, and macrophages.

[0511] According to one embodiment, in the indications of this application, the secretory gland cells include or are selected from at least one of the following: mammary gland cells, thyroid cells, and prostate cells.

[0512] According to one embodiment, in the indications of this application, the epithelial cells include or are selected from keratinocytes.

[0513] According to one embodiment, in the indications of this application, the lesion includes or is selected from tumors containing sensitive tumor cells.

[0514] According to one embodiment, in the indications of this application, the malignant tumor cells include or are selected from tumor cells that are not metformin cytotoxicity specific.

[0515] According to one embodiment, in the indications of this application, the tumor containing sensitive tumor cells is a tumor that cannot be effectively treated by standard metformin drugs, especially those with rapid onset. The drug or the use (or method) of this application can provide technical effects that standard metformin drugs or uses (or methods) cannot, including the type of action (transient vs. sustained action), the speed of action (rapid-acting vs. sustained-acting), and the method of administration required for the action (low-frequency vs. regular-frequency), to meet therapeutic needs that cannot be met by their existing uses (e.g., high efficacy, or even rapid onset).

[0516] According to one embodiment, in the indications of this application, the standard tumor is a tumor that cannot or can no longer be effectively treated by standard chemotherapy drugs.

[0517] According to one embodiment, in the indications of this application, X is the disease or symptom of the lesion that is not applicable or no longer applicable to standard treatment.

[0518] According to one embodiment, in the indications of this application, X is the disease or symptom that requires rapid treatment (not suitable for cases that do not show significant efficacy after a longer period of time, such as those with rapid disease progression or those requiring palliative care).

[0519] According to one embodiment, in the indications of this application, Y refers to the treatment, especially rapid-acting treatment, produced by the transient effect of the N,N-dimethylbiguanide solution on diseased tissue and its contained cells.

[0520] Within the scope of this invention, the term "tumor" refers to a pathological disease characterized by a lumpy mass, which can be a tumor of any pathology (malignant and non-malignant) and at any stage, including, for example, the following groups classified according to tumor cell type: epithelial cell tumors, sarcomas, lymphomas, germ cell tumors, germ cell tumors; and tumors named according to the organ or tissue in which the tumor cell concentration area is located, including, for example, tumors named according to the following organs or tissues: skin, bone, muscle, breast, kidney, liver, lung, gallbladder, pancreas, brain, esophagus, bladder, large intestine, small intestine, spleen, stomach, prostate, testicle, ovary, or uterus.

[0521] Specifically, the malignant tumors include, for example, breast cancer, pancreatic cancer, thyroid cancer, nasopharyngeal cancer, prostate cancer, liver cancer, lung cancer, intestinal cancer, oral cancer, esophageal cancer, stomach cancer, laryngeal cancer, testicular cancer, vaginal cancer, uterine cancer, ovarian cancer, sarcoma, etc.

[0522] 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, stomach tumors, nasopharyngeal tumors, laryngeal tumors, testicular tumors, vaginal tumors, uterine tumors, fallopian tube tumors, ovarian tumors, etc.

[0523] According to one embodiment, the malignant cells include or are selected from standard malignant cells that are ineffective on their own, such as cancer cells other than liver cancer cells and sarcoma cells.

[0524] According to one embodiment, the non-malignant cells include or are selected from at least one of connective tissue cells and organ tissue cells.

[0525] According to one embodiment, the connective tissue cells include fibroblasts.

[0526] According to one embodiment, the organ tissue cells include or are selected from at least one of the following: hepatocytes, mammary gland cells, thyroid cells, and skin cells.

[0527] According to one embodiment, the non-malignant lesions described in the indications of this application include or are selected from at least one of the following groups: benign tumors, nodules.

[0528] In this application, the term "nodule" refers to a benign lesion other than a benign tumor, including, for example, hyperplasia (such as hyperplasia of the breast, thyroid, parathyroid, prostate, etc.), cysts, abnormal venous masses (such as hemorrhoids), local inflammatory swelling, and swelling caused by microbial infection. Hemorrhoids include internal hemorrhoids, external hemorrhoids, and mixed hemorrhoids.

[0529] In one embodiment, the benign lesion includes localized inflammation, particularly refractory inflammation. Within the scope of this invention, the term "localized inflammation" refers to non-neoplastic inflammation at a localized site, including, for example, alterative inflammation, exudative inflammation, and proliferative inflammation, which can be any suitable type known to those skilled in the art, such as one or more of the following: arthritis, mastitis, pancreatitis, thyroiditis, prostatitis, hepatitis, pneumonia, enteritis, stomatitis, pharyngitis, periodontitis, esophagitis, gastritis, gastric ulcer, rhinitis, sinusitis, laryngitis, tracheitis, bronchitis, vaginitis, endometritis, salpingitis, oophoritis, etc.

[0530] In one embodiment, the benign lesion includes skin diseases, particularly refractory skin diseases. Within the scope of this invention, the term "skin disease" refers to a lesion originating from or secondary to the skin or skin appendages, which can be any suitable lesion known to those skilled in the art, such as including one or more of the following: skin cancer, non-malignant tumors of the skin, 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 tinea), sexually transmitted diseases (e.g., syphilis, gonorrhea, and condyloma acuminata), allergic and autoimmune skin diseases (e.g., contact dermatitis, eczema, urticaria), physical skin diseases (e.g., photodermatitis, chilblains, corns, chapped hands and feet, pressure sores), connective tissue diseases (e.g., lupus erythematosus), pigmentary disorders (e.g., freckles, nevi, various spots), and diseases of skin appendages (e.g., acne, rosacea, seborrheic dermatitis, alopecia areata, hair loss, hyperhidrosis, and bromhidrosis).

[0531] The present invention provides a drug combination comprising metformin (R) and a component (Z), wherein the administration concentration (w / w) of metformin is 0.2%-51%, preferably 0.5%-50%, and the component comprises a cell-responsive antitumor drug (abbreviated as B).

[0532] In some embodiments, the drug combination can be used to treat lesions such as tumors or nodules in the subject.

[0533] In some embodiments, the tumor in the drug combination is a solid tumor suitable for administration to the tumor body.

[0534] In some embodiments, in the above-described drug combination, R provides specific activities that are difficult to provide by standard metformin (the active ingredient being a metformin salt, abbreviated as R'), including a specific effect of effectively reducing biological barrier function (abbreviated as activity A): 1) Application to the tumor; 2) Application rate: In the formula, nR represents the number of times R is applied to the tumor in one course of treatment, qR(i) represents the dosage of R in the i-th application (i=1, ..., nR), and: In the formula, nRi is the number of injection points for R at the i-th application, cR(ii) and vR(ii) are the concentration and volume of R applied at the ii-th injection point, respectively, wherein the concentration threshold of cR(ii) (W / W) is 0.1% or 0.2%, and the upper limit of the concentration threshold is 51%, and the volume threshold of vR(ii) is 1 / 800 or 1 / 300 of the target volume (abbreviated as vtarget(ii)), and the upper limit of the volume threshold is 3 times vtarget(ii); - The Z provides one of its key control steps (abbreviated as Activity B) through the following limitations, which enables its specific antitumor action across the tumor tissue barrier: 1) Intratumoral application and / or systemic application; 2) Application rate: In the formula, nZ and qB(j) represent the number of systemic applications of Z in one course of treatment and the dosage of the j-th application (j=1, ..., nB), respectively; nZ', cZ(j'), and vZ(j') represent the number of tumor-specific applications of Z in one course of treatment and the concentration and volume of the j'-th application (j'=1, ..., nZ'), respectively. - The type of combination is a synergistic combination in which the drug effects of activity A of R and activity B of B mutually enhance each other.

[0535] In some embodiments, in the above-described drug combination, the N,N-dimethylbiguanide (R) comprises <30%, preferably ≤5% metformin salt (R'), or does not include metformin salt (R'), and preferably, the N,N-dimethylbiguanide (R) is in the form of a powder for injection.

[0536] In some embodiments, in the above-described drug combination, the tumor is selected from tumors whose tissue barrier function is unfavorable to the inhibitory effect of Z on tumor cells, preferably refractory tumors selected from Z, including chemotherapy-resistant tumors, tumors with unfavorable microenvironments, tumors discontinued from antitumor drugs, tumors for which there are no effective chemotherapy drugs, and tumors carried by patients with contraindications or incompatibilities to standard chemotherapy drugs.

[0537] In some embodiments, in the above-mentioned drug combination, the tumor is selected from heterogeneous tumors, wherein the heterogeneity includes at least one of tissue heterogeneity, spatial heterogeneity, and safety heterogeneity, and the heterogeneous tumor is a tumor in which different tumors of the same type or different regions of the same tumor are set as different target areas for differentiated treatment based on the heterogeneity.

[0538] In some embodiments, in the above-described drug combination, component (Z) includes an antitumor drug selected from cellularly reactive drugs whose key control step is to cross the tumor tissue barrier.

[0539] In some embodiments, the cell-responsive drugs in the above-described drug combination include cytotoxic drugs and targeted antitumor drugs.

[0540] In some embodiments, the cytotoxic drugs in the above-described drug combination include DNA damaging agents, antimetabolites, microtubule inhibitors, and topoisomerase inhibitors.

[0541] In some embodiments, representative compounds of DNA-damaging agents in the above-described drug combination include cisplatin, carboplatin, oxaliplatin, ifosfamide, and doxorubicin.

[0542] In some embodiments, representative compounds of antimetabolites in the above-described drug combination include fluorouracil, gemcitabine, and methotrexate.

[0543] In some embodiments, representative compounds of the microtubule inhibitors in the above-described drug combination include paclitaxel and vincristine.

[0544] In some embodiments, the pharmaceutical agents in the above-described drug combination include or are selected from at least one of the following: cisplatin, cyclophosphamide, carboplatin, doxorubicin, ifosfamide, nitrogen mustard, fluorouracil, capecitabine, gemcitabine, methotrexate, pemetrexed, fludarabine, paclitaxel, docetaxel, vincristine, vinblastine, vindesin, etoposide, irinotecan, topotecan, daunorubicin, mitoxantrone, gefitinib, osimertinib, larotrectinib, pazopanib, trastuzumab and its derivatives.

[0545] In some embodiments, the application concentration (w / w) of dimethyl biguanide (R) in the drug combination can be 0.2%-51%, 0.2%-50%, 0.5-50%, 0.5-45%, 1%-45%, 5%-45%, 5%-40%, 5%-30%, 5%-25%, 5%-20%, 1%-30%, 1%-25%, 1%-20%, 0.5%-10%, 1%-5%, or ≤2%, such as 1%, 0.75%, or 0.5%.

[0546] In some embodiments, in the above-described drug combination, the dimethyl biguanide (R) and component (Z) can be administered simultaneously and sequentially, preferably at certain time intervals.

[0547] In some embodiments, in the above-described drug combination, the dimethyl biguanide (R) and component (Z) may be administered intratumorally, or the dimethyl biguanide (R) may be administered intratumorally and a portion of component (Z) may be administered intratumorally, while the other portion of component (Z) may be administered intraperitoneally, for example, or the dimethyl biguanide (R) may be administered intratumorally and component (Z) may be administered intraperitoneally, for example.

[0548] In some embodiments, in the above-described drug combination, the dimethyl biguanide (R) and component (Z) may be administered simultaneously or sequentially to the tumor, or the dimethyl biguanide (R) and a portion of component (Z) may be administered simultaneously or sequentially to the tumor, while another portion of component (Z) may be administered simultaneously or sequentially to the system, for example, intraperitoneally, or the dimethyl biguanide (R) may be administered to the tumor while the component (Z) may be administered to the system, for example, intraperitoneally or gastrointestinally.

[0549] In some embodiments, in the above-described drug combination, the dimethyl biguanide (R) and component (Z) may be administered sequentially to the tumor, or the dimethyl biguanide (R) and a portion of component (Z) may be administered sequentially to the tumor, while the other portion of component (Z) may be administered sequentially systemically, for example, intraperitoneally, or component (Z) may be administered systemically, for example, intraperitoneally, before or after the administration of dimethyl biguanide (R) to the tumor.

[0550] In some embodiments, both dimethyl biguanide (R) and component (Z) in the drug combination are administered to the tumor (combination relationship I); R and Z are administered to the tumor and systemically, respectively (combination relationship II); or R and a portion of BZ are administered to the tumor, and another portion of Z is administered systemically (combination relationship III).

[0551] The present invention also provides a pharmaceutical combination comprising formulation I and formulation II, wherein formulation I comprises N,N-dimethylbiguanide (R) or N,N-dimethylbiguanide (R) and component (Z), and formulation II comprises Z but does not contain R, wherein: 1) In formulation I, the N,N-dimethylbiguanide (R) comprises ≤5% or does not comprise a salt of N,N-dimethylbiguanide; 2) The method of sharing formulation I and formulation II is as follows: Formulation I is for intratumoral administration only and cannot be used for systemic administration. Formulation II can be used for intratumoral and / or systemic administration; 3) The combined amount of Formulation I and Formulation II is limited by the combined amount of Z and R: (1). The application rate of R is: In the formula, nR represents the number of times R is applied to the tumor in one course of treatment, qR(i) represents the dosage of R in the i-th application (i=1, ..., nR), and: In the formula, nRi is the number of application points, such as needle insertion points, of R at the i-th application, cR(ii) and vR(ii) are the concentration and volume of R applied at the ii-th application point, such as needle insertion point, respectively, where vR(ii) ≤ 3 times the target volume of application point ii (abbreviated as vtarget(ii)); (2). The application rate of Z is: In the formula, nZ and qZ(j) are the number of times Z is applied to the system in one course of treatment and the amount of Z applied in the jth time (j=1, ..., nB), respectively. nZ', cZ(j'), and vZ(j') are the number of times Z is applied to the tumor in one course of treatment and the concentration and volume of Z at the j'th time (j'=1, ..., nZ') application, respectively.

[0552] In some embodiments, in the above-described drug combination, component Z comprises or is selected from at least one of the following: cisplatin, cyclophosphamide, carboplatin, doxorubicin, ifosfamide, nitrogen mustard, fluorouracil, capecitabine, gemcitabine, methotrexate, pemetrexed, fludarabine, paclitaxel, docetaxel, vincristine, vinca, vindesin, etoposide, irinotecan, topotecan, daunorubicin, mitoxantrone, gefitinib, osimertinib, larotrectinib, pazopanib, trastuzumab and its derivatives.

[0553] In some embodiments, in the above-described drug combination, drug C includes or is selected from at least one of the following: sodium chloride, sodium bicarbonate, sodium dihydrogen phosphate, sodium lactate, and sodium acetate.

[0554] In some embodiments, in the above-described drug combination, the drug E includes or is selected from at least one of the following: biological macromolecular immunomodulators such as immune checkpoint inhibitors, cytokines, and TLR agonists; immune cell immunomodulators; and vaccine immunomodulators such as BCG.

[0555] In some embodiments, in the above drug combinations, the contraindication for mixing R is any one of the following groups: 1) A salt of N,N-dimethylbiguanide (R'); 2) An acidifying agent that significantly converts R into R', wherein the acidifying agent includes acidic pH adjusters and other acidic substances; 3) Standard metformin drug excipients with a dosage ratio (Q excipient / QR) >3.0 or >4.0, including their standard sustained-release carriers, such as gel carriers and nanocarriers; 4) Adjuvants that are unstable in strongly alkaline aqueous solutions, including, for example, reducing sugars, cellulose, liposomes, carbomer, and polyethylene glycol-modified nanocarriers.

[0556] The aforementioned mixing taboo refers to the need to avoid mixing with it throughout the entire process of formulation, preparation, product manufacturing, and use.

[0557] In some embodiments, the sharing method in the above-described drug combination includes: 1) R and Z are mixed for intratumoral administration. 2) R and Z are used for intratumoral administration, respectively. 3) R and Z are used for intratumoral administration and systemic administration, respectively. 4) R and some Z are mixed or used separately for intratumoral administration, while the other Z are used for systemic administration.

[0558] In some embodiments, the shared threshold in the above drug combination is 0.5%, 1%, 3.3%, 5%, or 10%.

[0559] According to the present invention, a pharmaceutical composition comprising any of the above-described pharmaceutical combinations of the present invention (the composition of the present invention) and a formulation comprising any of the above-described pharmaceutical combinations or compositions according to the present invention (hereinafter referred to as the R combination formulation of the present invention) are provided.

[0560] According to the present invention, N,N-dimethylbiguanide (R) and component (Z) are provided for use in medicaments, drug combinations or kits for treating tumors or nodules (hereinafter referred to as R combination therapeutic use 1 of the present invention).

[0561] According to the present invention, the use of the above-described pharmaceutical combinations or compositions of the present invention in the preparation of a medicament, pharmaceutical combination or kit for treating lesions such as tumors or nodules in a subject is provided (hereinafter referred to as R combination therapy use 2 of the present invention).

[0562] In some embodiments, in the R combination therapy use of the present invention described above, the application concentration (w / w) of the dimethyl biguanide is ≤51%, ≤50%, ≤40%, ≤30%, ≤25%, ≤20%, preferably ≤10%, preferably ≤5%, more preferably ≤2%, such as 1%, 0.75%, or 0.5%.

[0563] In some embodiments, the application concentration (w / w) of metformin (R) in the R combination therapy of the present invention is 0.2%-51%, 0.5-50%, 1%-40%, 5%-50%, 5%-40%, 5%-30%, 5%-25%, 5%-20%, 1%-30%, 1%-25%, 1%-20%, preferably 0.5%-10%, preferably 1%-5%, more preferably ≤2%, such as 1%, 0.75%, or 0.5%.

[0564] In some embodiments, the concentration (w / w) of the N,N-dimethylbiguanide salt in the R combination therapeutic use of the present invention is ≤30%, preferably ≤20%, preferably ≤10%, more preferably ≤5%, ≤4%, or ≤3%, more preferably ≤2%, particularly preferably ≤1%, such as 1% or 0.5%, and most preferably 0, that is, the R of the drug combination does not contain the N,N-dimethylbiguanide salt.

[0565] In some embodiments, the R composition of the present invention comprises N,N-dimethylbiguanide and optionally a solvent, the composition not containing any additives required to meet the safety requirements for cell culture or standard injection, the concentration of N,N-dimethylbiguanide in the composition being ≥ its interventional concentration, wherein the administration or interventional concentration (w / w) is 0.5% ≤ CA ≤ 51%, preferably 1-51%, 2-50%, 3-50%, 4-50%, 5-50%, 6-50%, 7-50%, 8-50%, 9-50%. %, 10-50%, 15-50%, 20-50%, 25-50%, 30-50%, 35-50%, 40-50%, 1-40%, 5-50%, 10-40%, 15-40%, 20-40%, 25-40%, 30-40%, 35-40%, or 1-35%, 5%-30%, etc., or any value and range within these ranges, such as 1.5%, 2%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7.7.5%, etc. 8.8.5%, 9.9.5%, 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, 16.5%, 17%, 17.5%, 18%, 18.5%, 19%, 19.5%, 20%, 21%, 21.5%, 22%, 22.5%, 23%, 23.5%, 24%, 24.5%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 45%, 50%.

[0566] In some embodiments, the composition comprises a solvent, such as a pharmaceutically acceptable carrier, preferably water.

[0567] In some embodiments, the composition does not include a salt of N,N-dimethylbiguanide.

[0568] In some embodiments, the concentration (w / w) of the N,N-dimethylbiguanide salt contained in the composition is ≤25%, preferably ≤10%, preferably ≤5%, preferably ≤4%, more preferably ≤3%, more preferably ≤2%, particularly preferably ≤1%, and most preferably 0, that is, the composition does not contain the N,N-dimethylbiguanide salt.

[0569] In one embodiment, the additive includes an acidic pH adjuster.

[0570] In one embodiment, the pH adjuster includes or is selected from hydrochloric acid, sulfuric acid, acetic acid, citric acid, phosphoric acid, malic acid, tartaric acid, lactic acid, carbonic acid, etc.

[0571] In some embodiments, the pharmaceutical combination or composition used above includes component Z, which includes or is selected from at least one of the following: cisplatin, cyclophosphamide, carboplatin, doxorubicin, ifosfamide, nitrogen mustard, fluorouracil, capecitabine, gemcitabine, methotrexate, pemetrexed, fludarabine, paclitaxel, docetaxel, vincristine, vinca, vindesin, etoposide, irinotecan, topotecan, daunorubicin, mitoxantrone, gefitinib, osimertinib, larotrectinib, pazopanib, trastuzumab and its derivatives.

[0572] In some embodiments, the pharmaceutical combination or composition used in the above-described applications includes drug C, which includes or is selected from at least one of the following: sodium chloride, sodium bicarbonate, sodium dihydrogen phosphate, sodium lactate, and sodium acetate.

[0573] In some embodiments, the pharmaceutical combination or composition used in the above-described applications includes drug E, which includes or is selected from at least one of the following: biological macromolecular immunomodulators such as immune checkpoint inhibitors, cytokines, and TLR agonists; immune cell immunomodulators; and vaccine immunomodulators such as BCG.

[0574] According to the present invention, a method for treating cells of lesions such as tumors or nodules (hereinafter referred to as the R combination treatment method of the present invention) is provided, comprising applying or intervening the above-described drug combination or composition of the present invention into the lesion, penetrating the lesion tissue, and contacting the cells within the tissue.

[0575] According to the present invention, a method for treating tumors or nodules (hereinafter referred to as the R combination treatment method 1 of the present invention) is also provided, comprising: applying the above-described drug combination and composition of the present invention to the tumor or nodule, preferably, the application comprising applying the preparation of the present invention to the tumor or nodule, and / or applying the metformin to the tumor or nodule and applying the drug intratumorally or systemically.

[0576] According to the present invention, a method for treating tumors in a subject (hereinafter referred to as the R combination treatment method 2 of the present invention) is provided, comprising administering the above-described drug combination or composition of the present invention to the subject.

[0577] In the above-mentioned R combination therapy, the drug combination or composition includes dimethyl biguanide and optionally a solvent, wherein the formulation does not contain an acidic pH adjuster required to meet the safety requirements of cell culture or standard injection, wherein the concentration of N,N-dimethyl biguanide is ≥ its interventional concentration, wherein the interventional concentration (w / w) is 0.5% ≤ C ≤ 51%, preferably 1%-50%, more preferably 3%-40% or 5%-35%.

[0578] In some embodiments, the drug combination or composition may contain a solvent, preferably water, more preferably water for injection.

[0579] In some embodiments, the drug combination or composition may not include metformin salt.

[0580] In some embodiments, the concentration (w / w) of the dimethyl biguanide in the drug combination or composition can be ≤51%, ≤50%, ≤40%, ≤30%, ≤25%, ≤20%, preferably ≤10%, preferably ≤5%, more preferably ≤2%, such as 1%, 0.75% or 0.5%.

[0581] In some embodiments, the concentration (w / w) of metformin(R) in the drug combination or composition may be 0.2%-51%, 0.2%-50%, 0.5-50%, 1%-40%, 5%-50%, 5%-40%, 5%-30%, 5%-25%, 5%-20%, 1%-30%, 1%-25%, 1%-20%, preferably 0.5%-10%, preferably 1%-5%, or preferably ≤2%, such as 1%, 0.75%, or 0.5%.

[0582] In some embodiments, the acidic pH adjuster may include or be selected from hydrochloric acid, sulfuric acid, acetic acid, citric acid, phosphoric acid, malic acid, tartaric acid, lactic acid, and carbonic acid.

[0583] In some embodiments, in the above method, R is contained in a powder.

[0584] In some embodiments, in the above method, the powder consists of a dry powder containing R and a solvent.

[0585] In some embodiments, in the above method, the dry powder is R dry powder, and the solvent is water for injection.

[0586] In some embodiments, the pharmaceutical combination or composition does not contain an acid that would cause N,N-dimethylbiguanide(R) in the aqueous solution to be significantly converted into metformin salt.

[0587] In the above-mentioned R combination therapy or R combination treatment method, the applied drug combination or composition comprises metformin (R) and component (Z), wherein the applied concentration of metformin is 0.2%-51% by weight / w / w, preferably 0.5%-50%, and the component comprises a cell-responsive antitumor drug (abbreviated as B).

[0588] In some embodiments, in the above-described R combination therapy method or R combination treatment method, the drug combination includes: - The R provides activities (abbreviated as Activity A) that standard metformin (the active ingredient being a metformin salt, abbreviated as R') cannot provide, including effectively reducing biological barrier function (abbreviated as Activity A1): 1) Application to the tumor; 2) The dosage for one course of treatment is: In the formula, nR represents the number of times R is applied to the tumor in one course of treatment, qR(i) represents the dosage of R in the i-th application (i=1, ..., nR), and: In the formula, nRi is the number of application points of R at the i-th application, such as the number of injection points, cR(ii) and vR(ii) are the concentration and volume of R applied at the ii-th application point, such as the injection point, respectively, wherein the concentration threshold of cR(ii) (w / w) is 0.1% or 0.2%, and the upper limit of the concentration threshold is 51%, and the volume threshold of vR(ii) is 1 / 800 or 1 / 300 of the target volume (abbreviated as vtarget(ii)), and the upper limit of the volume threshold is 3 times vtarget(ii); - The cell-responsive antitumor drug (B) provides the cell response (abbreviated as active B) by means of the following, one of the key controlled steps of which is crossing the aforementioned biological barrier: 1) Intratumoral application and / or systemic application; 2) The dosage for one course of treatment is: In the formula, QB is the standard dosage of B in tumors sensitive to B, nB and qB(j) are the number of systemic administrations of B in one course of treatment and the dosage of the j-th administration (j=1, ..., nB), respectively, and nB', cB(j'), and vB(j') are the number of tumor administrations of B in one course of treatment and the concentration and volume of the j'-th administration (j'=1, ..., nB'), respectively. - The type of combination is a synergistic combination in which the drug effects of activity A of R and activity B of B mutually enhance each other.

[0589] In some embodiments, the N,N-dimethylbiguanide (R) comprises <30% (w / w), preferably ≤5% metformin salt (R'), or does not include metformin salt (R'), and preferably, the N,N-dimethylbiguanide (R) is in the form of a powder for injection.

[0590] In some embodiments, the component (Z) in the drug combination includes an antitumor drug selected from cell-reactive drugs (B) whose key control step is to cross the tumor tissue barrier.

[0591] In some embodiments, the cell-responsive drug (B) includes cytotoxic drugs and targeted antitumor drugs.

[0592] In some embodiments, the cytotoxic drug includes DNA damaging agents, antimetabolites, microtubule inhibitors, and topoisomerase inhibitors.

[0593] In some embodiments, representative compounds of the DNA damaging agent include cisplatin, carboplatin, oxaliplatin, ifosfamide, and doxorubicin.

[0594] In some embodiments, representative compounds of the antimetabolite include fluorouracil, gemcitabine, and methotrexate.

[0595] In some embodiments, representative compounds of the microtubule inhibitor include paclitaxel and vincristine.

[0596] In some embodiments, the targeted antitumor drug includes a small molecule targeted antitumor drug, preferably a kinase inhibitor.

[0597] In some embodiments, the kinase inhibitors include gefitinib, osimertinib, larotrectinib, pazopanib, trastuzumab, and their derivatives.

[0598] In some embodiments, in the above-described combination therapy R, the component Z comprises or is selected from at least one of the following: cisplatin, cyclophosphamide, carboplatin, doxorubicin, ifosfamide, nitrogen mustard, fluorouracil, capecitabine, gemcitabine, methotrexate, pemetrexed, fludarabine, paclitaxel, docetaxel, vincristine, vinblastine, vindesin, etoposide, irinotecan, topotecan, daunorubicin, mitoxantrone, gefitinib, osimertinib, larotrectinib, pazopanib, trastuzumab and its derivatives.

[0599] In some embodiments, in the above-described R combination treatment method, the drug C includes or is selected from at least one of the following: sodium chloride, sodium bicarbonate, sodium dihydrogen phosphate, sodium lactate, and sodium acetate.

[0600] In some embodiments, in the above-described R combination therapy, the drug E includes or is selected from at least one of the following: biological macromolecular immunomodulators such as immune checkpoint inhibitors, cytokines, and TLR agonists; immune cell immunomodulators; and vaccine immunomodulators such as BCG.

[0601] In some embodiments, in the above-described R combination therapy, the application concentration (w / w) of dimethyl biguanide (R) can be 0.2%-51%, 0.2%-50%, 0.5-51%, 0.5-50%, 0.5-45%, 1%-45%, 5%-45%, 5%-40%, 5%-30%, 5%-25%, 5%-20%, 1%-30%, 1%-25%, 1%-20%, 0.5%-10%, 1%-5%, or ≤2%, such as 1%, 0.75%, or 0.5%.

[0602] In some embodiments, in the above-described R combination therapy, the drug combination includes bimethyl biguanide (R) and doxorubicin (DOX), wherein the bimethyl biguanide (R) can be administered at a concentration of 0.2%-50% (w / w), and the doxorubicin (DOX) can be administered at a concentration of 0.025%-1% (w / w). Preferably, the bimethyl biguanide (R) is administered at a concentration of 5%-50% (w / w), and the doxorubicin (DOX) is administered at a concentration of 0.1%-1%. Optionally, both bimethyl biguanide (R) and doxorubicin (DOX) are administered to the tumor.

[0603] In one embodiment, metformin (R) is administered to the tumor at a concentration of 0.2%-50% (w / w), doxorubicin (DOX) is administered to the tumor at a concentration of 0.025%-1% (w / w), and doxorubicin (DOX) is administered systemically at a concentration of 0.05%-0.06% (w / w), such as intraperitoneally.

[0604] In some embodiments, in the above-described R combination treatment method, the drug combination includes dimethylbiguanide (R) and ifosfamide (IFO), wherein the dimethylbiguanide (R) can be administered at a concentration of 0.2%-50% (w / w), and the ifosfamide (IFO) can be administered at a concentration of 0.1%-10% (w / w). Preferably, the dimethylbiguanide (R) is administered at a concentration of 0.2%-45% (w / w), and the ifosfamide (IFO) is administered at a concentration of 0.1%-10% (w / w). More preferably, the dimethylbiguanide (R) is administered at a concentration of 5%-45% (w / w), and the ifosfamide (IFO) is administered at a concentration of 0.5%-10% (w / w). Optionally, both dimethylbiguanide (R) and ifosfamide (IFO) are administered to the tumor.

[0605] In one embodiment, the dimethyl biguanide (R) is administered to the tumor at a concentration of 5%-50% (w / w), the ifosfamide (IFO) is administered to the tumor at a concentration of 0.5%-10% (w / w), and the ifosfamide (IFO) is administered systemically at a concentration of 0.5% (w / w), such as intraperitoneally or gastrointestinally.

[0606] In some embodiments, in the above-described R combination therapy, the drug combination includes bimethyl biguanide (R) and cisplatin (DDP), wherein the bimethyl biguanide (R) can be administered at a concentration (w / w) of 0.2%-50%, and the cisplatin (DDP) can be administered at a concentration (w / w) of 0.01%-0.4%. Preferably, the bimethyl biguanide (R) is administered at a concentration (w / w) of 0.5%-50%, and the cisplatin (DDP) is administered at a concentration (w / w) of 0.01%-0.04%. Optionally, both bimethyl biguanide (R) and cisplatin (DDP) are administered to the tumor.

[0607] In one embodiment, the dimethyl biguanide (R) is administered to the tumor at a concentration of 0.5%-50% (w / w), the cisplatin (DDP) is administered to the tumor at a concentration of 0.013%-0.4% (w / w), and the cisplatin (DDP) is administered to the tumor systemically at a concentration of 0.015% (w / w), such as intraperitoneally.

[0608] In some embodiments, in the above-described R combination therapy, the drug combination includes dimethylbiguanide (R) and 5-fluorouracil (5-FU), wherein the dimethylbiguanide (R) can be administered at a concentration (w / w) of 0.2%-50%, and the 5-fluorouracil (5-FU) can be administered at a concentration (w / w) of 0.05%-9%. Preferably, the dimethylbiguanide (R) is administered at a concentration (w / w) of 0.2%-45%, and the 5-fluorouracil (5-FU) is administered at a concentration (w / w) of 0.5%-9%. More preferably, the dimethylbiguanide (R) is administered at a concentration (w / w) of 5%-45%, and the 5-fluorouracil (5-FU) is administered at a concentration (w / w) of 0.9%-9%. Optionally, both dimethylbiguanide (R) and 5-fluorouracil (5-FU) are administered to the tumor.

[0609] In one embodiment, the dimethyl biguanide (R) is administered to the tumor at a concentration of 5%-45% (w / w), the 5-fluorouracil (5-FU) is administered to the tumor at a concentration of 0.9%-9% (w / w), and the 5-fluorouracil (5-FU) is administered systemically at a concentration of 0.5% (w / w), such as intraperitoneally.

[0610] In some embodiments, in the above-described R combination therapy, the drug combination includes bimethyl biguanide (R), cisplatin (DDP), and gemcitabine (GEM), wherein the bimethyl biguanide (R) can be administered at a concentration of 0.2%-50% (w / w), the cisplatin (DDP) can be administered at a concentration of 0.05%-1% (w / w), and the gemcitabine (GEM) can be administered at a concentration of 0.5-2% (w / w), preferably at a concentration of 1% (w / w). Optionally, bimethyl biguanide (R) and cisplatin (DDP) can be administered to the tumor and gemcitabine (GEM) can be administered systemically, such as intraperitoneally.

[0611] In some embodiments, in the above-described R combination therapy, the drug combination includes dimethyl biguanide (R), paclitaxel (PTX), and 5-fluorouracil (5-FU), wherein the dimethyl biguanide (R) can be administered at a concentration of 0.2% to 50% (w / w), the 5-fluorouracil (5-FU) can be administered at a concentration of 0.1% to 10% (w / w), and the paclitaxel can be administered at a concentration of 0.02% to 1% (w / w), preferably at a concentration of 0.03% (w / w). Optionally, the dimethyl biguanide (R) and 5-fluorouracil (5-FU) are administered to the tumor and the paclitaxel (PTX) is administered systemically, such as intraperitoneally.

[0612] In some embodiments, in the above-described R combination therapy, the drug combination includes bimethyl biguanide (R) and osimertinib, wherein the bimethyl biguanide (R) can be administered at a concentration of 0.2%-50% (w / w), and the osimertinib can be administered at a concentration of 0.05%-2% (w / w). Preferably, the bimethyl biguanide (R) is administered at a concentration of 5% (w / w), and the osimertinib is administered at a concentration of 0.15% (w / w). Optionally, the bimethyl biguanide (R) is administered via vomiting, and the osimertinib is administered systemically, such as intraperitoneally.

[0613] In some embodiments, in the above-described R combination therapy, the drug combination includes bimethyl biguanide (R) and gefitinib (GR), wherein the bimethyl biguanide (R) can be administered at a concentration of 0.2% to 50% (w / w), and the gefitinib (GR) can be administered at a concentration of 0.1% to 2% (w / w). Preferably, the bimethyl biguanide (R) is administered at a concentration of 1% to 50% (w / w), and the gefitinib (GR) is administered at a concentration of 0.5% (w / w). Optionally, the bimethyl biguanide (R) is administered to the tumor, and the gefitinib (GR) is administered systemically, for example, intraperitoneally.

[0614] In some embodiments, in the above-described R combination therapy, the drug combination includes dimethyl biguanide (R), cisplatin (DDP), and gefitinib (GR), wherein the dimethyl biguanide (R) is administered at a concentration of 5% (w / w) (e.g., in the tumor), the gefitinib (GR) is administered at a concentration of 0.5% (w / w) (systemically, e.g., intraperitoneally), and the cisplatin (DDP) is administered at a concentration of 0.15% (w / w) (e.g., in the tumor).

[0615] In some embodiments, in the above-described R combination therapy, the drug combination includes dimethyl biguanide (R), ifosfamide (IFO), and gefitinib (GR), wherein the dimethyl biguanide (R) is administered at a concentration of 5% (w / w) (e.g., in the tumor), the ifosfamide (IFO) is administered at a concentration of 1% (w / w) (e.g., in the tumor), and the gefitinib (GR) is administered at a concentration of 0.5% (w / w) (systemically, e.g., intraperitoneally).

[0616] In some embodiments, in the above-described R combination therapy, the drug combination includes dimethyl biguanide (R), doxorubicin (DOX), and gefitinib (GR), wherein the dimethyl biguanide (R) is administered at a concentration of 5% (w / w) (e.g., in the tumor), the doxorubicin (DOX) is administered at a concentration of 0.05% (w / w) (e.g., in the tumor), and the gefitinib (GR) is administered at a concentration of 0.5% (w / w) (systemically, e.g., intraperitoneally).

[0617] In some preferred embodiments, in the above-described R combination treatment method, the contact, application, or intervention may include micro-volume multi-point contact, application, or intervention. Preferably, micro-volume multi-point injection, wherein the micro-volume for lesions larger than 3 cm includes amounts from 5 μL to 1000 μL, such as 5 μL-500 μL, 5 μL-250 μL, 5 μL-200 μL, 5 μL-100 μL, 5 μL-75 μL, 5 μL-50 μL, 5 μL-50 μL, etc. The dosage ranges are μL-25μL, 5μL-20μL, 10μL-200μL, 10μL-100μL, 10μL-75μL, 20μL-200μL, 20μL-100μL, 20μL-75μL, 30μL-200μL, 30μL-100μL, 40μL-200μL, 40μL-100μL, 50μL-200μL, 50μL-100μL, or 50μL-75μL, etc. Preferably, the dosage is applied through micro-volume multi-point contact or administration; more preferably, it is administered via micro-volume multi-point injection.

[0618] In some implementations, the above treatment can be sustained for at least one course of treatment. The duration of one course of treatment can be approximately 7 to 40 days, for example, approximately 9 to approximately 37 days, for example, approximately 15, 20, 25, 27, 28, 29, 30, 31, 32, 33, 34, or 35 days.

[0619] In one embodiment, during a course of treatment, contact with or application of the formulation of the present invention may be at a low frequency, such as about 1-6 times per course of treatment, preferably 1-5 times per course of treatment, 1-4 times per course of treatment, 1-3 times per course of treatment, or 1-2 times per course of treatment.

[0620] In some embodiments, in the above-described R combination therapy, the dimethyl biguanide (R) and component (Z) in the drug combination can be administered simultaneously, sequentially, or at intervals.

[0621] In some embodiments, in the above-described R combination therapy, both dimethyl biguanide (R) and component (Z) in the drug combination are administered to the tumor (combination relationship I); R and Z are administered to the tumor and systemically, respectively (combination relationship II); or R and a portion of Z are administered to the tumor, and another portion of Z is administered systemically (combination relationship III).

[0622] In some embodiments, in the above method, the tumor is a solid tumor suitable for tumor application.

[0623] In some embodiments, in the above-described R combination therapy, the tumor is selected from tumors whose tissue barrier function is unfavorable to the inhibitory effect of Z on tumor cells, preferably refractory tumors selected from Z, including non-indication tumors, drug-resistant tumors, tumors with unfavorable microenvironment, and tumors that have discontinued treatment.

[0624] In some embodiments, in the above-described R combination treatment method, the tumor is selected from heterogeneous tumors, wherein the heterogeneity includes at least one of tissue heterogeneity, spatial heterogeneity, and safety heterogeneity. The heterogeneous tumor is a tumor in which different tumors of the same type or different regions of the same tumor are designated as different target areas for differentiated treatment based on the heterogeneity.

[0625] In some embodiments, in the above-described R combination treatment method, the contact or application includes multi-point contact or application, preferably micro-volume multi-point contact or application, more preferably micro-volume multi-point injection.

[0626] In some embodiments, in the above-described R combination therapy, the formulation or drug combination is suitable for application to microvolume administration or intervention to reduce (Y) lesion volume (X), with the ratio of the administered volume to the target volume (V_administered / V_target) being approximately 0.02-0.056, 0.02-0.10, or 0.02-0.34.

[0627] In some embodiments, in the above-described R combination treatment method, when the lesion is ≥3.5 cm3 or the maximum size is ≥2 cm, the number of points is ≥5 points, such as 10, 15, 20, 25, 30, 35 or 50 points.

[0628] In some embodiments, in the above-described R combination treatment method, the contact or application is low-frequency, preferably 1-6 times per treatment course, more preferably 1-5 times per treatment course, 1-4 times per treatment course, 1-3 times per treatment course, or 1-2 times per treatment course.

[0629] In some embodiments, in the above-mentioned R combination treatment method, preferably, the application concentration CR of R is ≥5%, ≥10%, ≥20%, or ≤30%; and the number of application points is >3, 10, 15, 20, 25, 30, 35, or 100.

[0630] In some embodiments, in the above-described R combination therapy, the metformin R and component Z may be provided by one or more formulations, and wherein: R and Z are administered intratumorally; R and Z refer to intratumoral administration and systemic administration, respectively; or The R and part of the Z are administered intratumorally, and another part of the Z is administered systemically.

[0631] In some embodiments, in the above-mentioned R / Z combination therapy, the combination of R and Z (R / Z combination) is a synergistic combination that exceeds the expected effect of the standard metformin drug (R') and Z combination (R' / Z combination). This is manifested in that when the treatment dose of R (QR) is ≤25% or 50% of the treatment dose of R' (QR') and the treatment dose of Z is not increased, the drug effect is significantly increased, including at least one increase of at least 100% in drug adjustability indicators and at least one enhancement of at least 25% in drug efficacy indicators. The drug adjustability refers to the ability of the drug to provide differentiated administration parameters to different target areas while still maintaining efficacy.

[0632] In some embodiments, in the above-described R / Z combination therapy, the R / Z combination is also a synergistic combination that exceeds the expected effects of the HSA or Bliss method based on R monotherapy and Z monotherapy, characterized by an increased drug effect when the dosage of R (QR) is not increased and the dosage of Z (QB) is reduced by at least 20%, including an increase of at least one drug adjustability indicator by at least 10% and an enhancement of at least one drug efficacy indicator by at least 10%.

[0633] In some embodiments, in the above-described R or R / Z combination treatment methods, the drug adjustability index includes at least one of the following local application parameter adjustability coefficients (the ratio of the highest value in the adjustable range to its threshold): 1) A concentration-adjustable coefficient of R, which is the ratio of the highest value of cR to the threshold (highest value of cR / threshold), wherein the increase in the adjustable index is the increase in the coefficient, preferably the coefficient is increased to >10, 50, 100 or 150, which allows R to provide highly variable concentrations to different regions while maintaining its stable activity, including, for example, individualized drug administration for heterogeneous tumors; 2) The target volume adjustable coefficient of R is the ratio of the highest value of the target volume (v_target) to the threshold (highest value of v_target / threshold), and the increase of the adjustable index is the increase of the coefficient, preferably the coefficient is increased to >2, 10, 50, or 100, which allows R to select different v_targets for different treatment situations, including, for example, individualized drug administration for heterogeneous tumors; 3) The adjustable volume ratio coefficient of R, which is the ratio of the highest value of the ratio of the administered volume (vR) to its target volume (vtarget) to a threshold (vR / vtarget maximum value / threshold), wherein the increase in the adjustable index is the increase in the coefficient, preferably the coefficient is increased to >5, 10, or 100, which allows R to provide a highly variable volume ratio to different regions while still maintaining its stable activity, including, for example, individualized drug delivery for heterogeneous tumors; 4) The single-dose adjustment coefficient of R, which is the ratio of the highest value of the single-dose (cRi×vRi) to the threshold (highest value of cRi×vRi / threshold), wherein the increase in the adjustable index is the increase in the coefficient, preferably the coefficient is increased to >2, 6, or 10, which allows R to be selected with different single-dose doses for different treatment situations, including, for example, individualized treatment for heterogeneous tumors; and 5) The dosing point density adjustable coefficient of R is the ratio of the highest value of the dosing point density (nii / v target) to the threshold (highest value of nii / v target / threshold), and wherein the increase in the adjustable index is the increase in the coefficient, preferably the coefficient is increased to >3, 10, 50, or 100, which allows R to provide highly variable densities to different regions while maintaining its activity stability, including, for example, individualized dosing for heterogeneous tumors.

[0634] In some embodiments, in the above-described R or R / Z combination therapy, the drug efficacy indicator includes at least one of the following: 1) Time-effectiveness indicator, which is the time to significant effect in mouse experiments (the day when the tumor proliferation rate is <42%), and the enhancement of the efficacy indicator is the reduction in the time to significant effect. This makes the regimen a rapid-acting regimen, especially suitable for tumors that require rapid reduction in tumor size; 2) Efficacy indicators, expressed as tumor inhibition rate or objective response rate in mouse experiments, wherein the enhancement of the efficacy indicator is the increase in the tumor inhibition rate or objective response rate, which makes the regimen a highly effective regimen, especially suitable for tumors that need to be suppressed as much as possible in terms of tumor size; 3) Long-acting indicator, which is the progression-free time in mouse trials, wherein the enhancement of the efficacy indicator is the extension of the progression-free time, which makes the regimen a long-acting regimen, especially suitable for tumors that require the extension of the progression-free time as much as possible.

[0635] According to the present invention, a kit is provided comprising the formulation of the present invention as described above, and instructions for carrying out the methods described above.

[0636] According to the present invention, a kit is provided comprising a drug combination or composition as described above, and instructions for carrying out the treatment or treatment method as described above.

[0637] In one embodiment, the kit further includes a micro-volume intervention device, preferably suitable for micro-volume multi-point contact or application, and preferably includes a micro-injection device such as a syringe.

[0638] In one embodiment, the volumetric intervention device may include, for example, a puncture needle, an injection needle, a catheter, or other devices with the same function.

[0639] According to the present invention, the drug of this application is a therapeutic drug. When used to treat diseased lesions, it can be used as the main therapeutic drug and can also be used in combination with other interventional therapies, systemic chemotherapy, immunotherapy, photodynamic therapy, sonodynamic therapy, surgical intervention, or combinations of such therapies to further improve the therapeutic effect.

[0640] Beneficial effects of the present invention

[0641] First, it should be noted that because R can only be applied locally and not systemically like R', this application's method is limited to patients requiring application to the tumor site. The treatment of these patients still needs to be addressed, although tumor-based application currently accounts for less than 1% of clinical cases. The beneficial effects of this method for these patients are as follows:

[0642] 1. Comparison of the present invention with prior art metformin: The active ingredient in the formulation of the present invention is N,N-dimethylbiguanide (R), while the active ingredient in the prior art is metformin salt (R'). During the transient period between local application and conversion to R', R exhibits an active form distinct from the latter, demonstrating a dose-reduction synergistic effect exceeding the expectations of the prior art (R and R' are pharmaceutically equivalent). When the dosage of R in a single course of treatment (QR) is less than 25%-50% of the dosage of R' (QR'), the drug effect is significantly increased, including an increase of at least one drug adjustability indicator by at least 100% and an enhancement of at least one drug efficacy indicator by at least 40%. The efficacy indicators include time to onset of action, tumor inhibition rate or objective response rate, and time to no progression. This improved efficacy allows R to address the following issues that R' cannot: rapid onset, high efficacy, and / or long-lasting effect. The adjustability metrics include: maximum / threshold value of cR, maximum / threshold value of vtarget, maximum / threshold value of vR / vtarget, maximum / threshold value of cRi×vRi, and maximum / threshold value of nii / vtarget. Improved adjustability enables R to address the following issues that R' cannot: providing highly variable administration parameters to different tumor regions while maintaining stable efficacy, including, for example, individualized administration to heterogeneous tumors.

[0643] 2. Comparison of the present invention with existing technologies for transient local drugs (e.g., anhydrous ethanol): 1) The soluble concentration of R in the present invention (e.g., 51%) / threshold concentration (e.g., 0.2% or 0.5%) is tens of times higher, solving the long-standing problem of low adjustability indices for such drugs. 2) The transient local action of R in the present invention has better complementarity with the local action mechanisms of other drugs (e.g., methylene blue), providing better synergistic effects. 3) The efficacy-to-toxicity ratio of R in the present invention is higher, and the efficacy-to-toxicity ratio of combinations containing R and C (e.g., SB) is further greatly improved, thus providing a better technical solution for patients sensitive to local irritation.

[0644] 3. Comparison of the present invention with the prior art of metformin (R') and B combined (R' / B): In the combination of R and B in this scheme, the aforementioned transient activity of R, even in tumor tissue where it cannot completely induce necrosis, can provide a reduction in biological barrier function that R' cannot provide. This is beneficial for Z to overcome this key control step, which in turn is beneficial for the development of R's efficacy in this region. The combination of R and B thus provides a synergistic effect that exceeds the expectations of the prior art (R / B and R' / B are pharmaceutical equivalents), manifested in the realization of the beneficial effect of R in 1 above (the dose reduction synergistic effect), and at least the efficacy indicators such as tumor inhibition rate and progression-free time are further improved compared to R due to the combination with B. Even compared with R monotherapy and cell-reactive antitumor drugs (B), the present invention, at least in B-refractory tumors, further enhances the efficacy of the combination compared to the strongest monotherapy, manifested in at least a 10% increase in at least one of the drug adjustability indicators and / or at least a 10% increase in at least one of the drug efficacy indicators.

[0645] 4. Comparison of this invention with existing technologies using metformin (R') and E in combination (R' / E): In this scheme, the transient activity of R in the combination of R and E provides an effective release of pathogenic immune substances and inflammatory signals, even in tumor tissue where it cannot completely cause necrosis. Combined with the aforementioned effect of effectively reducing biological barrier function, it promotes immune cell infiltration. These effects provide an important basis and synergistic mechanism for the action of E. The combination of R and the immunomodulator (E) provides a synergistic effect that exceeds the expectations of existing technologies (R / E and R' / E are pharmaceutically equivalent), resulting in a more than 50% increase in the beneficial effects of R / E (e.g., long-term survival rate) compared to R' / E. Even compared with R monotherapy and E monotherapy, this combination further enhances the efficacy of the most potent monotherapy, resulting in a at least 25% increase in long-term survival rate.

[0646] Based on the research described in more detail below, although the specific mechanism requires further investigation, R in the drug of this application exhibits a transient effect, especially a transient penetration effect on the structure of diseased tissue. This effect enables the drug solution to reach the cells in the tissue and cause effective morphological changes, ultimately leading to effective destruction or reduction in the volume of the diseased tissue, and also causing direct damage to tumor cells.

[0647] Other objects, advantages, and novel features of the invention will become apparent to those skilled in the art upon examination of the following embodiments, which are not intended to be limiting. Furthermore, each of the various embodiments and aspects of the invention as described above and claimed in the claims section finds experimental support in the following embodiments.

[0648] Example

[0649] The present invention will be further illustrated by the following specific embodiments, but these are not intended to limit the invention. In the following embodiments, all experimental animals were tested in accordance with relevant regulations and industry self-regulation.

[0650] Unless otherwise specified, the cells, materials, reagents, etc. used in the following specific embodiments are all commercially available.

[0651] In the following embodiments, unless otherwise stated, all cells used were purchased from West China Hospital of Sichuan University and its wholly-owned subsidiary (Sichuan Kangcheng Biotechnology Co., Ltd.), and the drug resistance index of the drug-resistant cells was 4 or higher; the tumor tissue of the patient-derived xenograft model (hereinafter referred to as PDX) and its drug resistance verification were provided by West China Hospital of Sichuan University, and related experiments were conducted in cooperation with West China Hospital.

[0652] In the following examples, the tissue barrier function reducing agent studied was N,N-dimethylbiguanide (abbreviated as R, CAS 657-24-9), and the comparative agent was metformin hydrochloride (abbreviated as R', CAS 1115-70-4). Both raw materials were purchased from Shanghai Maclean Biochemical Technology Co., Ltd. The N,N-dimethylbiguanide dry powder used was prepared by removing hydrochloric acid from metformin hydrochloride, according to the supplier. Its purity quality standard was ≥95% (quality inspection result was 95.02%). It was tested to be a sterile powder. The highest concentration at which no precipitation was observed after dissolving under rapid stirring at room temperature (25°C) and standing for 2 hours was 51%. Therefore, in this application, 51% (w / w) represents the maximum solubility concentration of N,N-dimethylbiguanide in aqueous solution.

[0653] In the following examples, a sufficient number of representative cell-responsive drugs (abbreviated as B) were selected to fully demonstrate that the following results can be reasonably extended to this class of drugs: the tumor tissue barrier function is reduced by the same compound (R), which is beneficial to the antitumor response of any representative drug at the tumor cell level. The representative drugs of B were selected from cytotoxic drugs (B1) and targeted antitumor drugs (B2), and their raw materials were purchased from Shanghai Maclean Biotechnology Co., Ltd. and Shanghai Weihuan Biotechnology Co., Ltd.

[0654] In the following examples, representative cytotoxic drugs are selected from DNA damaging agents, antimetabolites, and microtubule inhibitors. Representative compounds of DNA damaging agents include: cisplatin (abbreviated as DDP, CAS15663-27-1), carboplatin (abbreviated as CBP, CAS41575-94-4), ifosfamide (abbreviated as IFO, CAS3778-73-2), and doxorubicin (abbreviated as DOX, CAS23214-92-8); representative compounds of antimetabolites include: fluorouracil (abbreviated as 5-FU, CAS51-21-8), gemcitabine (abbreviated as GEM, CAS95058-81-4), and methotrexate (abbreviated as MTX, CAS59-05-2); representative compounds of microtubule inhibitors include: paclitaxel (abbreviated as PTX, CAS33069-62-4) and vincristine (abbreviated as VCR, CAS57-22-7).

[0655] In the following examples, representative drugs for targeted antitumor therapy are selected from kinase inhibitors, etc. Representative compounds of kinase inhibitors include: gefitinib (GR, 184475-35-2), osimertinib (AZD9291, CAS1421373-65-0), larotrectinib (CAS223403-58-4), pazopanib (CAS444731-52-6), and trastuzumab (CAS180288-69-1).

[0656] In the following examples, the immunomodulator (E) is selected from one or more of the following: biological macromolecular immunomodulators (abbreviated as E1), immune cell immunomodulators (abbreviated as E2), and vaccine immunomodulators (abbreviated as E3). E1 is selected from immune checkpoint inhibitors including Anti-mouse PD-1 (CD279)-InViVo (Selleck, abbreviated as RMP1-14), cytokines including interleukin-15 (abbreviated as IL-15, CAS150801-17-9), and TLR agonists including CpG ODN 1826 (abbreviated as CpG). Vaccine immunomodulators include BCG (Chengdu Institute of Biological Products).

[0657] In the following embodiments, methylene blue (CAS615-731-6) is abbreviated as MB.

[0658] In the following examples, unless otherwise stated, all test animals were purchased from Chengdu Dashuo Biotechnology Co., Ltd., and animal experiments were conducted in the company's animal laboratory. Balb / c nude mice, Balb / c mice, and C57BL / 6 mice were healthy animals aged 5-6 weeks, with normal weight (18-22g), normal diet (not fasted), and no diseases (including no metabolic system diseases or diabetes). Subcutaneous xenograft tumor formation was performed according to the testing guidelines issued by the drug regulatory authority (e.g., CFDA "Guidelines for Pharmacodynamics of Antitumor Drugs"). Unless otherwise stated, xenograft tumors were modeled subcutaneously in the right axilla of the animals. Unless otherwise stated, once the lesion reached the desired volume (e.g., 100-300 mm3), animals were randomly divided into several groups of 6 animals each using PEMS 3.2 software. Intratumoral injection was performed to ensure the drug was distributed as evenly as possible within the tumor. The experimental observations, measurements, and analyses included general condition, weight, food intake, lesion volume, tumor weight, etc. Unless otherwise specified, no abnormalities were observed in general condition, weight, or food intake. Animal experiments involving subcutaneous transplanted nodules were conducted according to the methods described above for animal experiments involving subcutaneous transplanted tumors.

[0659] In the following examples, experimental results (e.g., tumor weight) are expressed as mean ± standard error (x+SEM). Differences between the means of the two experimental animal groups were compared using the statistical software The SPSSAU project (2023) (SPSSAU. (version 23.0) [Online Application Software]. Retrieved from https: / / www.Spssau.com), with a significance level of 0.05.

[0660] The formula for calculating tumor volume is as follows: Tumor volume (V) = 1 / 2 × a × b², where a represents tumor length and b represents tumor width. The formula for calculating relative tumor volume (RTV) is as follows: RTV = Vt / V0, where Vt and V0 are the tumor volumes on the study day (t) and the randomization day, respectively.

[0661] The formula for calculating the relative tumor proliferation rate (T / C%) is as follows: T / C% = [(Tt-T0) / (Ct-C0)] × 100, where Tt, T0 and Ct, C0 are the tumor volumes of the study group and the negative control group on study day (t) and randomization day, respectively.

[0662] The formula for calculating the tumor inhibition rate is as follows: Tumor inhibition rate Y(%) = (CW-TW) / CW×100%, where TW is the average tumor weight in the study group and CW is the average tumor weight in the negative control group.

[0663] In the following embodiments, the evaluation criteria for various drug effects are as described above.

[0664] Example 1, R Study: Using drug-resistant tumors as a lesion model

[0665] In this embodiment, the standard metformin-resistant lesion model (hereinafter referred to as refractory lesion) is a tumor resistant t...

Claims

1. A formulation comprising N,N-dimethylbiguanide and a solvent, wherein the formulation does not contain an acidic pH adjuster required to meet the safety requirements for cell culture or standard injection, wherein the concentration of N,N-dimethylbiguanide in the formulation is ≥ its application concentration, wherein the application concentration (w / w) is 0.5% ≤ CR ≤ 51%, and wherein the concentration (w / w) of metformin salt in the formulation is ≤ 5%.

2. A formulation specifically for interventional treatment of lesions, comprising N,N-dimethylbiguanide and a solvent, wherein the formulation does not contain an acidic pH adjuster required to meet the safety requirements of cell culture or standard injection, wherein the concentration of N,N-dimethylbiguanide is ≥ its interventional concentration, wherein the interventional concentration (w / w) is 0.5% ≤ C ≤ 51%, and wherein the concentration (w / w) of metformin salt in the formulation is ≤ 2%.

3. The formulation as claimed in claim 1 or 2, wherein the solvent is water.

4. The formulation as claimed in claim 1 or 2, wherein the formulation does not include a salt of N,N-dimethylbiguanide and an acid that causes the N,N-dimethylbiguanide to be converted into a metformin salt.

5. The formulation as claimed in claim 1 or 2, wherein the concentration (w / w) of the N,N-dimethylbiguanide in the formulation is 1-50%.

6. The formulation as claimed in claim 1 or 2, wherein the concentration (w / w) of the N,N-dimethylbiguanide in the formulation is 1%-40%.

7. The formulation as claimed in claim 1 or 2, wherein the acidic pH adjuster or acid includes hydrochloric acid, sulfuric acid, acetic acid, citric acid, phosphoric acid, malic acid, tartaric acid, lactic acid, and carbonic acid.

8. The formulation as claimed in claim 1 or 2, wherein the formulation comprises N,N-dimethylbiguanide (R) and water for injection.

9. The formulation as claimed in claim 1 or 2, wherein the concentration (w / w) of the metformin salt in the formulation is ≤1%.

10. A pharmaceutical composition for treating localized lesions, comprising N,N-dimethylbiguanide (abbreviated as R) and a component (Z), wherein the N,N-dimethylbiguanide is administered at a concentration (w / w) of 0.1%-51%, and the component (Z) comprises at least one selected from the group consisting of: cell-reactive antitumor drugs (abbreviated as B), near-neutral or weakly basic sodium salts (abbreviated as C), methylene blue dyes (abbreviated as D), and immunomodulators (abbreviated as E), wherein the formulation does not contain metformin salts or acids that convert the N,N-dimethylbiguanide to metformin salts.

11. The pharmaceutical combination as claimed in claim 10, wherein the N,N-dimethylbiguanide is defined by the following to provide an active form distinct from standard metformin (whose active ingredient is a metformin salt): 1) Topical application; 2) The dosage for one course of treatment is: QR = qR(i), where QR < 50% or 25% of the equivalent dosage of the metformin salt, nR is the number of times R is applied to the tumor in one course of treatment, qR(i) is the dosage of R in the i-th application (i = 1, ..., nR), and: qR(i) = cR(ii) × vR(ii), where nR is the number of application points of R in the i-th application, cR(ii) and vR(ii) are the concentration and volume of R applied at the ii-th application point, respectively, wherein the concentration threshold of cR(ii) (W / W) is 0.1% or 0.2%, and the upper limit of the concentration threshold is 51%, and the volume threshold of vR(ii) is 1 / 800 or 1 / 300 of the target volume (abbreviated as vtarget(ii)), and the upper limit of the volume threshold is 3 times vtarget(ii). The active form of metformin salt is characterized by complete complexation with acid in solution; while the active form of R is characterized by less complexation with acid in solution, mainly existing in a form of very little or partial complexation. Thus, through the transient local action of this different active form, it provides an activity that metformin salt cannot provide (abbreviated as activity A), including at least one of the following: the effect of effectively reducing the function of biological barriers (abbreviated as activity A1), the effect of effectively damaging diseased tissue (abbreviated as activity A2), and / or the effect of effectively releasing diseased immune substances and / or inflammatory signals within the lesion (abbreviated as activity A3).

12. The pharmaceutical combination as claimed in claim 10, which is a pharmaceutical combination for treating solid tumors, comprising said R and B, wherein: - The activity A of R includes at least effectively reducing the function of the biological barrier (activity A1); - The cell-reactive antitumor drug (B) provides the cell response (abbreviated as activity B) by means of the following, one of the key control steps of which is crossing the aforementioned biological barrier: 1) Tumor administration and / or systemic administration; 2) A treatment course dosage of: QB = qB(j) + cB(j') × vB(j'), where QB is the standard dosage ≤ in tumors sensitive to B, nB and qB(j) are the number of systemic administrations of B in a treatment course and the dosage of the jth (j=1, ..., nB) administration, respectively, and nB', cB(j'), and vB(j') are the number of tumor administrations of B in a treatment course and the concentration and volume of the j'th (j'=1, ..., nB') administration, respectively.

13. The pharmaceutical combination of any one of claims 10 to 12, comprising said R, C, and / or D, wherein: - The activity A of R includes at least the effect of effectively damaging diseased tissue (activity A2); - The C is defined as providing an effect that improves the local efficacy-toxicity ratio of R (abbreviated as activity C) by: 1) being mixed with R; 2) the mixing ratio of C to R (QC / QR) is 0.5 to 4.0; - The D is defined as providing a pharmacological effect that improves the drug effect of R by: 1) lesion application, including mixed application with R or separate application; 2) a treatment course dose QE of ≤ a transient effective dose for local action.

14. The pharmaceutical combination of any one of claims 10 to 12, comprising said R and E, wherein: - The activity A of R includes at least the effect of effectively reducing the function of biological barriers (activity A1) and / or the effect of effectively releasing lesion immune substances and / or inflammatory signals within the lesion (activity A3); - The activity E is defined by the following to provide immunomodulatory effects benefiting from the effects of activity A1, and / or the effect of activity A3 to optimize local and / or systemic immune responses (abbreviated as activity E): 1) lesion administration and / or systemic administration; 2) The dosage for one course of treatment is: QE = qB(k) + cE(k') × vE(k'), where QE is the standard dosage ≤ qB(k) in the anti-lesion application of E, nE and qE(k) are the number of systemic applications of E in one course of treatment and the dosage of the kth application (j=1, ...nk), respectively, and nE', cE(k') and vE(k') are the number of tumor applications of E in one course of treatment and the concentration and volume at the k'th application (k'=1, ...nk'), respectively.

15. The formulation as claimed in claim 1 or 2, wherein the formulation does not include metformin salt, and optionally, N,N-dimethylbiguanide (R) is in the form of a powder for injection.

16. The pharmaceutical combination of any one of claims 10 to 12, wherein the N,N-dimethylbiguanide (R) is in the form of a powder for injection.

17. The pharmaceutical combination of any one of claims 10 to 12, wherein the cell-responsive antitumor drug comprises at least one of the following: cisplatin, cyclophosphamide, carboplatin, doxorubicin, ifosfamide, nitrogen mustard, fluorouracil, capecitabine, gemcitabine, methotrexate, pemetrexed, fludarabine, paclitaxel, docetaxel, vincristine, vinblastine, vindesin, etoposide, irinotecan, topotecan, daunorubicin, mitoxantrone, gefitinib, osimertinib, larotrectinib, pazopanib, trastuzumab and derivatives thereof.

18. A composition comprising an formulation as described in any one of claims 1 to 9 or a pharmaceutical combination as described in any one of claims 10 to 16.

19. A formulation comprising a pharmaceutical combination as described in any one of claims 10 to 16 or a composition as described in claim 18.

20. Use of an N,N-dimethylbiguanide (abbreviated as R) as a transient active ingredient in the preparation of a medicament for treating tumors or nodules, wherein the N,N-dimethylbiguanide does not contain metformin salt or an acid that converts the N,N-dimethylbiguanide to metformin salt.

21. Use of an formulation as claimed in any one of claims 1 to 9 in the preparation of a medicament for treating a tumor or nodule in a subject.

22. The use as described in claim 20 or 21, wherein the N,N-dimethylbiguanide is used to provide a morphological manifestation distinct from that of standard metformin (the active ingredient of which is metformin salt), wherein the active form of the metformin salt is in a form that is completely complexed with acid in solution; while the active form of R is in a form that is less complexed with acid in solution, mainly existing in a form that is very little or partially complexed, thereby providing an activity (abbreviated as activity A) that is difficult for metformin salt to provide through the transient local action of this distinct active form, including the effect of effectively reducing the function of biological barriers (abbreviated as activity A1), the effect of effectively damaging diseased tissue (abbreviated as activity A2), and / or the effect of effectively releasing disease immune substances and / or inflammatory signals within the lesion (abbreviated as activity A3).

23. The use as claimed in claim 20 or 21, wherein the R is not mixed with a sufficient amount of metformin salt (R') or an acidifying agent capable of converting the R into a sufficient amount of metformin salt, wherein the sufficient amount means an amount such that the ratio of the metformin salt to R (R' / R) is ≥1 / 3.

24. The use as described in claim 20 or 21, wherein the N,N-dimethylbiguanide (R) is defined by the following to provide an active form distinct from standard metformin drugs (whose active ingredient is a metformin salt): 1) not being mixed with acids that would cause the R to be significantly converted to a metformin salt; 2) being applied topically to lesions; 3) ... The dosage for one course of treatment is: QR = qR(i), where QR < 50% or 25% of the equivalent dosage of the metformin salt, nR is the number of times R is applied to the lesion or tumor in one course of treatment, qR(i) is the dosage of R in the i-th application (i = 1, ..., nR), and: qR(i) = cR(ii) × vR(ii), where nRi is the number of application points of R in the i-th application, cR(ii) and vR(ii) are the concentration and volume of R applied at the ii-th application point, respectively, wherein the concentration threshold of cR(ii) (W / V) is 0.1% or 0.2%, and the upper limit of the concentration threshold is 51%; the volume threshold of vR(ii) is 1 / 800 or 1 / 300 of the target volume (abbreviated as vtarget(ii)), and the upper limit of the volume threshold is 3 times vtarget(ii). The active form of metformin salt is characterized by complete complexation with acid in solution; while the active form of R is characterized by less complexation with acid in solution, mainly existing in a form of very little or partial complexation. Thus, through the transient local action of this different active form, it provides an activity that metformin salt cannot provide (abbreviated as activity A), including at least one of the following: the effect of effectively reducing the function of biological barriers (abbreviated as activity A1), the effect of effectively damaging diseased tissue (abbreviated as activity A2), and / or the effect of effectively releasing diseased immune substances and / or inflammatory signals within the lesion (abbreviated as activity A3).

25. The use as claimed in claim 24, wherein R and drug (Z) are used together, wherein the application concentration (w / w) of R is 0.2%-50%, and said drug (Z) comprises at least one selected from the group consisting of: cell-responsive antitumor drugs (abbreviated as B), near-neutral or weakly basic sodium salts (abbreviated as C), methylene blue (abbreviated as D), and immunomodulators (abbreviated as E), wherein: - The cell-responsive antitumor drug (B) provides the cell response (abbreviated as active B) by means of the following defined criteria, one of the key control steps of which is to cross the aforementioned biological barrier: 1) lesion administration and / or systemic administration; 2) a treatment course dosage of: QB = qB(j) + cB(j') × vB(j'), where QB is ≤ the standard dosage in tumors sensitive to B, nB and qB(j) are the number of systemic administrations of B in a treatment course and the dosage of the jth (j=1, ..., nB) administration, respectively, and nB', cB(i'), and vB(i') are the number of tumor administrations of B in a treatment course and the concentration and volume at the j'th (j'=1, ..., nB') administration, respectively; - The C is used to provide an effect that improves the local efficacy-toxicity ratio of the R (abbreviated as active C) by means of the following defined criteria: 1) 1) Administered in combination with R; 2) The mixing ratio (QC / QR) of C and R is 0.5 to 4.0; - D provides efficacy to enhance the pharmacological effect of R by the following limitations: 1) Administered to the lesion, including in combination with or separately from R; 2) The treatment dose QE is ≤ a transient effective dose for local action. C is used to provide efficacy to enhance the local efficacy-toxicity ratio of R (abbreviated as active C) by the following limitations: 1) Administered in combination with R; 2) The mixing ratio (QC / QR) of C and R is 0.5 to 4.0; - D provides efficacy to enhance the pharmacological effect of R by the following limitations: 1) Administered to the lesion, including in combination with or separately from R; 2) The dosage QE for one course of treatment is ≤ a transient effective dosage for local action. The E is defined by the following methods to provide immunomodulatory effects benefiting from the effects of the active A1, and / or to optimize local and / or systemic immune responses by utilizing the effects of the active A3 (abbreviated as active E): 1) Pathological application and / or systemic application; 2) The dosage for one course of treatment is: QE = qB(k) + cE(k') × vE(k'), where QE is ≤ a standard dosage for the anti-pathological application of the E, nE and qE(k) are the number of systemic applications of E in one course of treatment and the dosage at the kth application (j=1, ..., nk), respectively, and nE', cE(k'), and vE(k') are the number of tumor applications of E in one course of treatment and the concentration and volume at the k'th application (k'=1, ..., nk'), respectively.

26. The use as described in claim 24, wherein the R is used in conjunction with a cell-responsive antitumor drug (abbreviated as B) and an immunomodulatory agent (abbreviated as E), wherein: - The cell-responsive antitumor drug (B) provides the cell response (abbreviated as Active B) by means of the following defined criteria, one of the key controlled steps of which is crossing the aforementioned biological barriers: 1) lesion administration and / or systemic administration; 2) ... The dosage for one course of treatment is: QB = qB(j) + cB(j') × vB(j'), where QB is the standard dosage ≤ in the sensitive tumors of B, nB and qB(j) are the number of systemic administrations of B in one course of treatment and the dosage of the jth administration (j=1, ..., nB), respectively, and nB', cB(i'), and vB(i') are the number of tumor administrations of B in one course of treatment and the concentration and volume at the j'th administration (j'=1, ..., nB'), respectively. The immunomodulator (E) is defined by the following methods for providing immunomodulatory effects benefiting from the effects of activity A1 and / or optimizing local and / or systemic immune responses using the effects of activity A3 (abbreviated as activity E): 1) lesion administration and / or systemic administration; 2) ... The dosage for one course of treatment is: QE = qB(k) + cE(k') × vE(k'), where QE is the standard dosage ≤ qB(k) in the anti-lesion application of E, nE and qE(k) are the number of systemic applications of E in one course of treatment and the dosage of the kth application (j=1, ..., nk), respectively, and nE', cE(k') and vE(k') are the number of tumor applications of E in one course of treatment and the concentration and volume at the k'th application (k'=1, ..., nk'), respectively.

27. The use as described in claim 20 or 21, wherein the R is contained in a powder.

28. The use as described in claim 25 or 26, wherein the cell-responsive antitumor drug (B) includes cytotoxic drugs and targeted antitumor drugs.

29. The use as claimed in claim 28, wherein the cytotoxic drug comprises a DNA damaging agent, an antimetabolite, or an antimicrotubule agent, wherein the DNA damaging agent comprises cisplatin, carboplatin, oxaliplatin, ifosfamide, and doxorubicin; the antimetabolite comprises fluorouracil, gemcitabine, and methotrexate; and the antimicrotubule agent comprises paclitaxel and vincristine.

30. The use as described in claim 28, wherein the targeted antitumor drug includes a kinase inhibitor.

31. The use as described in claim 30, wherein the kinase inhibitor comprises gefitinib, osimertinib, larotrectinib, pazopanib, trastuzumab and derivatives thereof.

32. The use as described in claim 25 or 26, wherein the near-neutral or weakly basic sodium salt (C) comprises: Sodium chloride, sodium bicarbonate (abbreviated as SB), sodium dihydrogen phosphate (MSP), sodium lactate (abbreviated as Nal), sodium acetate (abbreviated as SA).

33. The use as described in claim 25 or 26, wherein the near-neutral or weakly basic sodium salt (C) is sodium bicarbonate (SB).

34. The use as described in claim 25 or 26, wherein the methylene blue dye (D) includes methylene blue, patent blue, isothiocyanate blue and neomethylene blue.

35. The use as described in claim 25 or 26, wherein the methylene blue dye (D) is methylene blue.

36. The use as described in claim 25 or 26, wherein the immunomodulator (E) includes a biomolecular immunomodulator (abbreviated as E1), an immune cell immunomodulator (abbreviated as E2), and a vaccine immunomodulator (abbreviated as E3).

37. A kit comprising a formulation as described in any one of claims 1 to 9 or a combination of drugs as described in any one of claims 10 to 19.

38. The kit as claimed in claim 37, further comprising a microvolume intervention device.

39. The kit as claimed in claim 38, wherein the microvolume intervention device is suitable for micro-volume multi-point contact or application, wherein the microvolume is a drug application volume of ≤5% or 10% of the target volume.

40. The kit as claimed in claim 38 or 39, wherein the microvolume intervention device includes a microinjection device.

41. The use as described in claim 20 or 21, wherein the nodule is a nonmalignant nodule comprising at least one of the following nonmalignant cells: connective tissue cells, secretory gland cells, and epithelial cells.

42. The use as described in claim 20 or 21, wherein the tumor includes a solid tumor.

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