CN113230212A

CN113230212A - Aqueous emulsion comprising amino acids, use of such emulsion and pharmaceutical composition comprising such emulsion

Info

Publication number: CN113230212A
Application number: CN202010076627.0A
Authority: CN
Inventors: 邹方霖; 邹礼常; 王建霞; 王艺羲
Original assignee: Chengdu Kuachang Aopu Medical Technology Co ltd
Current assignee: Chengdu Kuachang Aopu Medical Technology Co ltd
Priority date: 2020-01-23
Filing date: 2020-01-23
Publication date: 2021-08-10

Abstract

The present disclosure relates to a stable water-based emulsion comprising an amino acid compound and an oil and fat, and in the emulsion, the concentration of the oil and fat is < 50% by weight, and the weight ratio of the oil and fat to the amino acid compound is < 8/1 or (1-320)/40, preferably < 4/1 or (1-160)/40. Also relates to the application and preparation method of the emulsion, and a pharmaceutical composition containing the emulsion.

Description

Aqueous emulsion comprising amino acids, use of such emulsion and pharmaceutical composition comprising such emulsion

Technical Field

The present disclosure relates to an aqueous-based emulsion comprising an amino acid-based compound, use of the emulsion and a pharmaceutical composition comprising the emulsion, a method of treating a solid tumor using the emulsion or the pharmaceutical composition, and a device for treating a solid tumor.

Background

Injectable oils can be used as liquid carriers (e.g., imaging carriers, embolization carriers, solvents for hydrophobic drugs, etc.) in the preparation of injectables for the treatment of diseases (e.g., solid tumors). However, injection dose limitations often limit the clinical application of these injectable oils. For example, a fat (e.g., an iodized oil) comprising a vegetable oil and a developing substance (e.g., iodine) may be used as the developer in an amount of about<25 ml. For another example, when vegetable oil is used as the embolizing carrier, the embolizing dose is also about<25 ml. To obtain more complete contrast or/and embolization, the amount of imaging agent or/and embolization agent must be increased, but the amount of oleaginous agent is too large and can cause more serious side effects (e.g., increased liver function damage). The addition of water with less side effects to dilute the oil and fat is beneficial to increase the injection dosage (for example, more than or equal to 50 ml). However, the mutual repulsion between the oil and water makes it difficult to form a stable emulsion system, and two-phase separation of the nonpolar liquid phase and the polar liquid phase occurs even before the injection is completed. For example, when an iodized oil-water mixed liquid is used as a developer, a high ratio (e.g., W) is often required_{Oil and fat}/W_{Water (W)}Not less than 1.0) to achieve effective emulsification so that the dosage is not greatly increased. For another example, when the iodized oil-water mixed liquid is used as a carrier of a conventional antitumor drug, a high proportion (for example ≥ 1.0) of oil is often required for effective emulsification, so that the dosage is not greatly increased. For another example, when an iodized oil-water mixed liquid is used as a carrier for a conventional antitumor agent, a high ratio (e.g., W) is often required_{Oil and fat}/W_{Water (W)}Not less than 1.0) to achieve effective emulsification and thus the increase in dosage is still not significant. As another example, when used as a paclitaxel solvent, high ratios are also requiredTo make a mixture with ethanol to further achieve emulsification with aqueous solutions at the time of use, a large proportion and thus a large volume of oil injection causes very severe hypersensitivity.

Thus, in order to improve the stability of injectable emulsions comprising oil and water or/and to reduce the concentration (dose) of oil that must be used, it is necessary to develop new emulsion enhancing techniques to meet this need that the prior art has not been able to meet.

Disclosure of Invention

According to one aspect of the present invention, there is provided a stable water-based emulsion comprising an amino acid-based compound and an oil or fat, and in the emulsion, the concentration of the oil or fat is < 50% by weight, and the weight ratio of the oil or fat to the amino acid-based compound is not more than 8/1 or (1-320)/40, preferably not more than 4/1 or (1-160)/40. The aqueous emulsion may be an injectable aqueous emulsion.

According to another aspect of the present invention there is provided the use of an aqueous-based emulsion according to the present invention for the preparation of a pharmaceutical composition for the treatment of solid tumors. In one embodiment, the pharmaceutical composition further optionally comprises one or more antineoplastic agents.

According to a further aspect of the present invention there is provided a pharmaceutical composition for the treatment of solid tumors comprising the water-based emulsion according to the present invention. In one embodiment, the pharmaceutical composition may also optionally comprise one or more antineoplastic agents.

According to a further aspect of the present invention there is provided a method of treating a solid tumour comprising the in vivo administration to a subject in need thereof of a therapeutically effective volume of an aqueous-based emulsion or pharmaceutical composition according to the present invention.

According to a further aspect of the present invention there is provided a method of preparing an aqueous emulsion according to the present invention comprising the steps of: 1) preparing an aqueous solution comprising the amino acid compound and optionally other water-soluble components; 2) preparing an oil solution comprising lipids and optionally other lipophilic components; and 3) thoroughly mixing the aqueous solution prepared in step 1) with the oil solution prepared in step 2).

According to a further aspect of the present invention, there is provided a process for the preparation of a pharmaceutical composition according to the present invention, comprising the steps of: 1) preparing an aqueous solution comprising the amino acid-based compound, optionally a water-soluble antineoplastic agent and optionally other water-soluble components; 2) preparing an oil solution comprising lipids, optionally lipophilic antineoplastic agents and optionally other lipophilic components; 3) mixing the aqueous solution prepared in step 1) with the oil solution prepared in step 2) thoroughly.

According to yet another aspect of the present invention there is provided a device for the treatment of solid tumours comprising a container a and a container B, wherein the container a contains an injectable aqueous solution, wherein the aqueous solution comprises the amino acid based compound and optionally other water soluble components in the required amounts according to formulation or dosing concentration; and the container B contains an injectable oil solution, wherein the oil solution comprises a desired amount of lipids and optionally other lipophilic components according to formulation or administration concentration; wherein the ratio of the aqueous solution to the oil solution is such that in the emulsion thus formed, the concentration of the oil is < 50% by weight, and the weight ratio of the oil to the amino acid-based compound is not more than 8/1 or (1-320)/40, preferably not more than 4/1 or (1-160)/40.

According to yet another aspect of the present invention there is provided a developer which is an aqueous based emulsion comprising an iodized oil and an amino acid based compound, wherein in the emulsion the concentration of said iodized oil is < 50% by weight and the weight ratio of said iodized oil to said amino acid based compound is < 8/1 or (1-320)/40 or preferably < 4/1 or (1-160)/40.

Compared with the prior art of medical injection emulsion, the water-based emulsion for injection provided by the invention has the following advantages: the amino acid compound under the preferable conditions can obviously improve the oil phase-water phase dispersion stability of the emulsion under the condition of low oil concentration. For example, it shows a significantly increased (2-to 20-fold or more) available dose and thus a significantly increased development area, as compared with the technique using the existing emulsion as a development injection; compared with the prior art which utilizes the emulsion as the embolism injection, the injection shows that the available dosage is obviously improved (more than 2 times to 20 times), thereby obviously improving the embolism area and/or the embolism curative effect; compared with the technology of using the existing emulsion as an injection for developing, embolizing and/or pathological structure destruction drugs, the emulsion has the advantages of maximizing available dose, minimizing the dilution of the aqueous solution of the active ingredients by grease and the like, and can generate better drug effect; compared with the prior art of using emulsion as the injection of lipophilic drug (such as paclitaxel) liquid carrier, the injection preparation shows obviously improved injection concentration of the conventional antitumor drug and/or obviously reduced injection volume. The pharmaceutical composition comprising the water-based emulsion for injection according to the present invention has the following advantages compared to the prior art of the anti-tumor injection emulsion: the low oil concentration can obviously improve (more than 2 times to 20 times) the available dosage and can better retain the activity of the antitumor drug. In addition, the application and the composition are convenient to prepare and low in cost, and are particularly beneficial to leading the vast population who is difficult to bear high expense to enjoy safe and effective treatment.

Detailed Description

The inventors of the present invention have unexpectedly found, in an experiment for formulating an oil-water two-phase mixture emulsion, that the addition of an amino acid compound such as arginine significantly reduces the rate of oil-water two-phase separation under certain specific conditions. These specific conditions are not the component weight ratios of the amino acid-based compound (usually as an active stabilizer) in a conventional oil-water two-phase mixture emulsion, but are as defined below.

In one aspect of the present disclosure, there is provided a stable injectable aqueous emulsion comprising an amino acid compound and a fat. In one embodiment, the concentration of said oil or fat is<50% by weight. In one embodiment, the concentration of said oil or fat is<50% by weight. In one embodiment, the weight ratio (W) of the oil or fat to the amino acid compound_{Oil and fat}/W_{Amino acid compound}) Is less than or equal to 8/1 or (1-320)/40, preferably less than or equal to 4/1 or (1-160)/40.

In yet another aspect of the present disclosure, there is provided an injectable aqueous-based emulsion for the preparation of a medicament for the treatment of solid tumorsUse in a composition wherein the emulsion comprises water and oil. In one embodiment, the concentration of said oil or fat is<50% by weight. In one embodiment, the concentration of said oil or fat is<50% by weight. In one embodiment, the weight ratio (W) of the oil or fat to the amino acid compound_{Oil and fat}/W_{Amino acid compound}) Is less than or equal to 8/1 or (1-320)/40, preferably less than or equal to 4/1 or (1-160)/40. In one embodiment, the pharmaceutical composition further optionally comprises one or more antineoplastic agents.

In yet another aspect of the present disclosure, there is provided a pharmaceutical composition for treating a solid tumor, comprising an injectable aqueous-based emulsion, and wherein the aqueous-based emulsion comprises an amino acid-based compound and an oil. In one embodiment, the oil concentration (w/w) in the composition is<50 percent. In one embodiment, the weight ratio (W) of the oil or fat to the amino acid compound_{Oil and fat}/W_{Amino acid compound}) Is less than or equal to 8/1 or (1-320)/40, preferably less than or equal to 4/1 or (1-160)/40. In one embodiment, the pharmaceutical composition further comprises one or more antineoplastic agents.

In yet another aspect of the present disclosure, there is provided a method of treating a solid tumor comprising administering in vivo to a subject in need thereof an aqueous emulsion or a pharmaceutical composition comprising the aqueous emulsion in a volume effective for treatment, and wherein the aqueous-based emulsion comprises an amino acid compound and an oil. In one embodiment, the oil concentration (w/w) is<50 percent. In one embodiment, the weight ratio (W) of the oil or fat to the amino acid compound_{Oil and fat}/W_{Amino acid compound}) Is less than or equal to 8/1 or (1-320)/40, preferably less than or equal to 4/1 or (1-160)/40. In one embodiment, the pharmaceutical composition further comprises one or more antineoplastic agents.

In the context of the present disclosure, the term "oil" is used to indicate a pharmaceutically acceptable oil and lipophilic derivatives thereof, preferably selected from injectable oils and injectable lipophilic derivatives thereof as carried in the chinese, us or european official pharmacopoeia or guidelines. In the context of the present disclosure, the term "amino acid based compound" is used to refer to pharmaceutically acceptable amino acids and derivatives thereof, preferably amino acid based compounds selected from the group of mono-or compound amino acid based nutritional drugs as carried in the chinese, us or european official pharmacopoeia or guidelines.

In the context of the present disclosure, the term "injectable emulsion" is used to refer to an injectable formulation that is in the form of an emulsion when in use. In the context of this disclosure, the term "injection" is used to refer to a drug for injection into the body as required by the drug administration, which usually has a liquid (e.g., solution, emulsion, suspension, etc.) form when injected into the body, but may have different dosage forms before use (e.g., concentrated injection comprising concentrated solution and its diluent, powder injection comprising sterile powder and its liquefier, emulsion injection comprising oil solution and aqueous solution, suspension injection comprising liquid phase component and microparticle component, etc.). The term "emulsion" or "emulsion" is used to refer to a two-phase mixed liquid comprising an oil solution and an aqueous solution, one of which is dispersed in the other in the form of droplets. The term "oil solution" is used to refer to a homogeneous liquid comprising one or more lipids. The term "aqueous solution" is used to refer to a homogeneous liquid comprising water.

In the context of the present disclosure, the term "intratumorally administering" is used to refer to the injection of a liquid drug (e.g., an injection) into a tumor via a device, such as transcatheter arterial infusion, transcatheter intratumoral infusion, intratumoral injection, and the like.

In the context of the present disclosure, unless otherwise indicated, the term "concentration" refers to the weight percent concentration (w/w) of a given component in a given composition. In the context of this disclosure, the term "formulation concentration" refers to the concentration of a given component in a form in which all components of the drug are well mixed, e.g., when the formulation is a drug concentrate, the concentration of the given component in a form in which it is completely diluted prior to administration. In the context of the present disclosure, the term "administration concentration" refers to the concentration of a given component in the form in which the drug is administered (e.g., as an injection into a syringe). In the context of the present disclosure, the term "initial target concentration" refers to the concentration of a given component in a drug-containing medium (e.g., drug-containing blood) as the drug enters the target area. Even if the concentrations of the conventional and topical preparations are the same, they are likely to be administered at different concentrations. The conventional administration is mainly due to pharmaceutical (e.g. commercial factor) rather than pharmacological reasons, and the concentration of the preparation is far higher than the design of blood concentration, so that the administration after dilution (administration concentration is lower than the preparation concentration) is often preferred clinically. Topical administration is often done at a concentration that is known as the formulation concentration for pharmacological reasons. Furthermore, even if the concentrations of the conventional and topical drugs are the same, their initial concentrations at the target area are necessarily very different. The conventional drug is distributed in the blood and then enters the target region after undergoing complicated physical, chemical, biological processes (e.g., dilution of different drug components in the blood, different adsorption, different retention or even metabolism in certain organs, etc.), and the initial concentration (blood concentration) of the drug at the target region is far (more than thousand times) lower than the administered concentration. And the concentration of the drug administered locally (e.g., the concentration in the injection needle or the outlet of the perfusion tube) is actually the initial concentration at the target area. In the context of the present disclosure, unless otherwise indicated, the concentrations of the components are the concentrations in the pharmaceutical compositions of the invention, both as to their formulation and as to their administration, or as to their initial target concentration. For example, in all embodiments, the concentration administered is the initial concentration in the target region, or the concentration of the formulation.

In one embodiment, the amino acid based compound concentration (w/w) in the aqueous emulsion or pharmaceutical composition is > 3%, > 5%. In one embodiment, the amino acid compound concentration (w/w) in the aqueous emulsion or pharmaceutical composition is 5-50%.

In one embodiment, the concentration (w/w) of the lipid in the aqueous emulsion or pharmaceutical composition is 30% or less or 1-30%, preferably 20% or 1-20%, or 10% or 1-10%. In one embodiment, the concentration (w/w) of said oil or fat in said aqueous emulsion or pharmaceutical composition is preferably ≦ 15% or 0.5-15%, more preferably ≦ 15% or 0.5-15%, or ≦ 5% or 0.5-5%.

In one embodiment, the oil or fat may be selected from an oil for injection and a derivative thereof, wherein the oil for injection is preferably selected from one or more of the following: ethyl oleate, medium-chain (C6-10) fatty acid glyceride, isopropyl myristate, soybean oil, tea oil, olive oil, castor oil, peanut oil, cottonseed oil, corn oil, linseed oil, poppy seed oil and sesame oil. In one embodiment, the oil comprises iodized oil. In one embodiment, the iodine content of the iodized oil is 30-50% (w/w). In one embodiment, the iodized oil comprises a superliquefied iodized oil.

In one embodiment, the lipid is a liquid carrier for the aqueous emulsion or pharmaceutical composition.

In another aspect of the present disclosure, there is provided a developer which is an aqueous-based emulsion comprising an iodized oil and an amino acid compound, wherein in the emulsion the concentration of the iodized oil is < 50% by weight and the weight ratio of the iodized oil to the amino acid compound is < 8/1 or (1-320)/40 or preferably < 4/1 or (1-160)/40.

In the context of the present disclosure, the term "liquid carrier" is used to refer to a liquid carrier. In the context of the present disclosure, the term "carrier" is used to indicate a substance which does not itself have a significant chemico-pharmacological effect but which is capable of delivering a pharmaceutically active ingredient to a defined site in the body without significantly diminishing its activity, the carrier typically also having one or more of the following effects: (1) sustained release to increase the duration of action of the pharmaceutically active ingredient (e.g. sustained release carrier); (2) increasing the selectivity of the pharmaceutically active ingredient (e.g., targeting vectors); (3) reducing the toxicity of the active ingredient; (4) overcoming the difficult problems encountered in the formulation of pharmaceutical agents (e.g. co-dispersing carriers); (5) physical effects such as embolic effects (e.g. embolic carriers).

In one embodiment, the amino acid compound is selected from amino acid compounds that can be used as nutritional agents, preferably selected from one or more of the following amino acids and derivatives thereof, carried in pharmacopoeia: arginine, alanine, glutamic acid, glycine, histidine, lysine, valine, isoleucine, leucine, phenylalanine, threonine, proline, serine. In one embodiment, the amino acid compound is one or more selected from the group consisting of: arginine, glutamic acid hydrochloride, glycine, histidine, lysine hydrochloride, lysine, valine, isoleucine, leucine, serine, alanyl-glutamine. In one embodiment, the amino acid class compound includes arginine. In one embodiment, the amino acid based compounds include arginine and other amino acid based compounds selected from those available as nutraceuticals as loaded in pharmacopoeia. In one embodiment, the amino acid class compound includes arginine and serine. In one embodiment, the amino acid class compounds include arginine and glycine.

In one embodiment, the pharmaceutical composition further comprises one or more antineoplastic agents. In one embodiment, the antineoplastic agent is selected from an unconventional antineoplastic agent or/and a conventional antineoplastic agent. In one embodiment, the antineoplastic agent is selected from the group consisting of non-conventional antineoplastic agents. In one embodiment, the concentration (w/w) of the non-conventional anti-neoplastic drug is more than 0.25%, preferably 0.35-40%. In one embodiment, the antineoplastic agent is selected from conventional antineoplastic agents. In one embodiment, the concentration (w/w) of the conventional antitumor agent is more than 0.05%, 0.05-15%, preferably 0.5-10%. In one embodiment, the antineoplastic agents include non-conventional antineoplastic agents and conventional antineoplastic agents, and the concentration (w/w) of said non-conventional antineoplastic agents in said pharmaceutical composition is greater than 0.25%, preferably 0.35-40%; the concentration (w/w) of the conventional antitumor drug is more than 0.05 percent, 0.05-15 percent and preferably 0.5-10 percent.

In the context of the present disclosure, the term "conventional antineoplastic agent" is used to refer to compounds which exhibit tumor cell inhibition in cell experiments, and also exhibit tumor cell targeting by absorption in animal experiments to produce a tumor-inhibiting effect as expected, e.g., as approved by a drug administration (e.g., FDA) for use as a conventional chemotherapeutic agent against a particular solid tumor. In the context of the present disclosure, the term "non-conventional antineoplastic agent" is used to refer to a conventional ineffective drug that is locally effective, which typically requires administration under specific conditions (e.g., intratumoral administration at a certain target initial concentration). The term "conventional ineffective drug" used is distinguished from the term "conventional antitumor drug", and refers to a drug other than the conventional antitumor drug, specifically, a compound which shows or does not show a tumor cell inhibitory effect in a cell experiment, but does not show a tumor cell-targeting effect by absorption in an animal experiment to thereby produce an antitumor effect as expected.

In one embodiment, the antineoplastic agent comprises a non-conventional antineoplastic agent, and wherein the non-conventional antineoplastic agent is selected from one or more of the group consisting of: vital dye in a concentration (w/w) of 0.5-15% in the pharmaceutical composition; a strong base at a concentration of 0.5% or more, or 0.5-6%, preferably 0.5-1.5% or 1.5-6% in the pharmaceutical composition; a weak base at a concentration (w/w) in the pharmaceutical composition of 3.5-50%; a strong acid at a concentration of 0.5% or more, or 0.5-3%, preferably 0.5-1.5% or 1.5-3% in the pharmaceutical composition; a weak acid in a pharmaceutical composition at a concentration (w/w) of 0.5-50%.

In the context of the present disclosure, the term "vital dye" is used to refer to a pharmaceutically acceptable aromatic compound that selectively absorbs or reflects light of a particular wavelength at a target region, which may, for example, include one or more of the following organic dyes and derivatives thereof: methylene blue (including its hydrates), patent blue, isothio blue, toluidine blue, trypan blue, basic blue, eosin, basic fuchsin, crystal violet, gentian violet, neutral red, janus green B, safranin, bengal red, and the like.

In one embodiment, the non-conventional anti-tumor drug comprises one or more selected from vital dyes, and the concentration (w/w) of the vital dyes in the pharmaceutical composition is 0.5% or more, 0.5-15%. In one embodiment, the vital dye is one or more selected from the group consisting of: methylene blue, patent blue, isothio blue, bengal red. In one embodiment, the concentration (w/w) of methylene blue, patent blue, or/and isothio blue is 0.5-1.5%. In one embodiment, the concentration (w/w) of said bengal is between 5 and 15%.

In one embodiment, the non-conventional anti-neoplastic agent comprises one or more selected from the group consisting of strong bases, and the concentration of the strong base in the pharmaceutical composition is 0.8% or more, 0.8 to 4%, preferably 0.8 to 1.5% (when the strong base is not used in combination with a weak acid or/and a weak base) or 1.5 to 4% (when the strong base is used in combination with a weak acid or/and a weak base). In one embodiment, the strong base is one or more selected from the group consisting of: sodium hydroxide, potassium hydroxide, calcium hydroxide. In one embodiment, the strong base comprises sodium hydroxide.

In one embodiment, the non-conventional antitumor agent comprises one or more selected from weak bases, and the concentration of the weak base is 3.0% or more and 5% or more, and is 3.0 to 35% or 5 to 35%. In one embodiment, the weak base is one or more selected from the group consisting of: inorganic salts of polybasic weak acids, alkali metal salts of organic weak acids, and weak bases containing nitrogen. In one embodiment, the weak acid inorganic salt is selected from one or more of the following: sodium phosphate, sodium carbonate, potassium carbonate, borax, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium bicarbonate, potassium bicarbonate, calcium bicarbonate and sodium hydrosulfate, wherein the concentration of the polybasic acid inorganic salt is more than or equal to 3.0 percent or more than or equal to 5 percent and is 3.0-15 percent or 5-10 percent. In one embodiment, the weak acid inorganic salt is preferably sodium bicarbonate. In one embodiment, the alkali metal salt of an organic weak acid is selected from one or more of the following: potassium hydrogen phthalate, sodium acetate, sodium propionate, sodium butyrate, sodium malonate, sodium lactate, sodium citrate, sodium malate and sodium dodecyl sulfate, wherein the concentration of the organic weak acid alkali metal salt is more than or equal to 5.0 percent and is 5.0-15 percent or 5-20 percent. In one embodiment, the alkali metal salt of an organic weak acid is preferably sodium acetate. In one embodiment, the weak nitrogen-containing base is selected from one or more of the following: ammonia water, ammonia chloride, 2-aminoethanol, tromethamine, triethanolamine, tris, 2-aminoethanol, tromethamine, triethanolamine, meglumine and gluglucosylamine, wherein the concentration of the nitrogen-containing weak base is not less than 5.0 percent and is 5.0 to 35 percent or 15 to 50 percent. In one embodiment, the non-conventional antineoplastic agent comprises 3 ± 1% sodium hydroxide and 7 ± 2% sodium bicarbonate.

In one embodiment, the non-conventional antineoplastic agent comprises one or more strong acids, and wherein the concentration of said strong acid is 0.8% or more, 0.8-5%, preferably 0.8-1.5% (when said strong acid is not combined with a weak acid or/and a weak base) or 1.0-5.0% (when said strong acid is combined with a weak acid or/and a weak base). In one embodiment, the strong acid is one or more selected from the group consisting of: hydrochloric acid, sulfuric acid, nitric acid. In one embodiment, the strong acid is preferably hydrochloric acid.

In one embodiment, the non-conventional anti-neoplastic agent comprises one or more weak acids, and the concentration of the weak acid is 3.0% or more, or 5% or more, or 3.0 to 20%. In one embodiment, the weak acid is one or more selected from the group consisting of the following C1-10 aliphatic carboxylic acids having 0-3 hydroxyl groups: acetic acid, glycolic acid, propionic acid, malonic acid, butyric acid, succinic acid, lactic acid (2-hydroxypropionic acid), citric acid (2-hydroxy-1, 2, 3-tricarballylic acid), malic acid (2-hydroxysuccinic acid), tartaric acid, oxalic acid, gluconic acid. In one embodiment, the weak acid is acetic acid. In one embodiment, the non-conventional antineoplastic agent comprises 1.5 + 1% hydrochloric acid and 8 + 3% acetic acid.

In one embodiment, the anti-neoplastic agent comprises the conventional anti-neoplastic agent and the vital dye. In one embodiment, the antineoplastic agent comprises the conventional antineoplastic agent and one or more selected from the group consisting of: the strong base, the weak base, the strong acid, the weak acid. In one embodiment, the antineoplastic agent comprises the conventional antineoplastic agent, the vital dye, and one or more selected from the group consisting of: the strong base, the weak base, the strong acid, the weak acid.

In one embodiment, the conventional antineoplastic agent is selected from one or more of the following groups: drugs that disrupt the structure and function of DNA, drugs that interfere with the transcription of RNA that intercalate into DNA, drugs that interfere with DNA synthesis, drugs that affect protein synthesis. In one embodiment, the conventional antitumor agent is one or more selected from the group consisting of: uracil derivatives, cyclophosphamide, gemcitabine, epirubicin, antitumor antibiotics, teniposide, metal platinum complex and taxanes. In one embodiment, the conventional antitumor agent is preferably one or more selected from the group consisting of the following drugs and their analogous derivatives: doxorubicin hydrochloride, gemcitabine, cisplatin, 5-fluorouracil, alexancept, carboplatin, mitomycin, captopop, cyclophosphamide, doxorubicin, carmustine, camptothecin, paclitaxel, vincristine, elemene, docetaxel, fotemustine.

In one embodiment, the conventional antineoplastic agent is selected from water-soluble conventional antineoplastic agents. In one embodiment, the conventional antineoplastic agent is selected from oil-soluble conventional antineoplastic agents. In one embodiment, the conventional antitumor agent is a combination selected from the group consisting of a water-soluble conventional antitumor agent and an oil-soluble conventional antitumor agent. In one embodiment, the water-soluble conventional antineoplastic agent comprises one or more of the following groups: doxorubicin hydrochloride, gemcitabine, cisplatin, 5-fluorouracil, alexancept, carboplatin, mitomycin, captopop, cyclophosphamide. In one embodiment, the oil-soluble conventional antineoplastic agent comprises one or more of the following groups: adriamycin, carmustine, camptothecin, paclitaxel, vincristine, elemene, docetaxel and fotemustine.

In one embodiment, the pharmaceutical composition further comprises optionally other components. In one embodiment, the other components include one or more of: ethanol, 1, 2-propylene glycol and glycerol. In one embodiment, the additional component further comprises an additional carrier, such as a particulate carrier.

According to the preparation method of the present invention, the preparation of the aqueous emulsion of the present invention comprises at least the following steps: 1) preparing an aqueous solution comprising the amino acid based compound and optionally water soluble further components; 2) preparing an oil solution comprising the oil and optionally other components of the oil that are soluble in the oil; and 3) thoroughly mixing the aqueous solution prepared in step 1) with the oil solution prepared in step 2).

According to the preparation method of the present invention, the preparation of the pharmaceutical composition of the present invention comprises at least the following steps: 1) preparing an aqueous solution comprising the amino acid-based compound, the antineoplastic agent (if a water-soluble antineoplastic agent is selected) and optionally the water-soluble further components; 2) preparing an oil solution comprising the lipid, the antineoplastic agent (if an oil-soluble antineoplastic agent is selected) and optionally other components of the lipid-soluble; and 3) thoroughly mixing the aqueous solution prepared in step 1) with the oil solution prepared in step 2).

According to one embodiment disclosed herein, the powder injection for injection of the pharmaceutical composition of the present invention may be prepared by a method comprising the steps of: cooling the aqueous solution prepared from the pharmaceutical composition to 10-25 deg.C, lyophilizing, plugging, and capping to obtain sterilized dry powder; preparing a sterile aqueous vehicle comprising the water and the optional other components in the required amounts according to formulation or dosing concentration; and preparing a sterile oil solution containing the oil or fat and optionally other components in amounts required according to formulation or administration concentration. The sterile dry powder is preferably sterile freeze-dried powder, and the preparation method comprises the following steps: 1) preparing an aqueous solution comprising or not comprising the amino acid based compound but optionally other components; 2) sterilizing, filtering and packaging; 3) freeze drying; 4) and (5) plugging and capping. The freeze-drying process conditions include, for example: the pre-freezing condition is that the temperature is kept at minus 45 ℃ for 4 hours; sublimation drying condition is that the heating rate is 0.1 ℃/min, and the heating is kept for at least 10 hours when the temperature is raised to-15 ℃; the desorption drying conditions were 30 ℃ for 6 hours. When in use, the sterile dry powder of the powder injection for injection is redissolved in a sterile aqueous solvent containing or not containing amino acid compounds to form a redissolved liquid medicine, and then the redissolved liquid medicine is fully mixed with a sterile oil solution to obtain the emulsion. In use, the emulsion can be directly used as intravenous injection or can be directly used for intratumoral administration of liquid medicines after being diluted.

In the present disclosure, the pharmaceutical composition is preferably for use in the treatment of solid tumors by intravenous administration or/and intratumoral administration. In one embodiment, the injectable emulsion comprises an intravenous emulsion. In one embodiment, the injectable emulsion comprises an intratumoral administration emulsion. In one embodiment, the pharmaceutical composition is preferably for use in imaging or/and treating solid tumors by intratumoral administration.

Within the scope of the present disclosure, the term "tumor" refers to a mass formed due to abnormal proliferation of cells or mutated cells, including solid tumors. The term "solid tumor" refers to a tumor having a tumor body, which may be due to any pathology (malignant and non-malignant) and at any stage of the tumor, including for example the following groups classified by tumor cell type: epithelial cell tumors, sarcomas, lymphomas, germ cell tumors, blastomas; and tumors named as the organ or tissue in which the tumor cell foci are located, including, for example, tumors named as the following organs or tissues: skin, bone, muscle, breast, kidney, liver, lung, gall bladder, pancreas, brain, esophagus, muscle of the shoulder, large intestine, small intestine, spleen, stomach, prostate, emerald, ovary, or uterus.

Specifically, the malignant tumor includes, 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, and the like. The non-malignant tumor includes, for example, breast tumor, pancreatic tumor, thyroid tumor, prostate tumor, liver tumor, lung tumor, intestinal tumor, oral tumor, esophageal tumor, stomach tumor, nasopharyngeal tumor, laryngeal tumor, testicular cancer, vaginal cancer, uterine tumor, fallopian tube tumor, ovarian tumor, etc.

The pharmaceutical composition disclosed herein is a therapeutic pharmaceutical composition which, when used for imaging or/and treating solid tumors, can also be administered in combination with other interventions, systemic chemotherapy, immunotherapy, photodynamic therapy, sonodynamic therapy, surgical intervention or a combination of such therapies to further enhance the therapeutic effect. In one embodiment, the method comprises optionally further performing one or more other treatments, such as chemotherapy, immunotherapy, radiotherapy, surgery, physical ablation, before, during or after intravenous or/and intratumoral administration of the pharmaceutical composition.

In the use and method for imaging or/and treating solid tumors according to the present disclosure, the intratumorally administrable pharmaceutical composition comprising lipids, water, and amino acid compounds is administered in their concentration or amount ratio in the pharmaceutical composition disclosed herein. This concentration or amount provides better emulsification than administration, and also allows the treatment to be more effective.

In another aspect of the present disclosure, there is provided a device for treating a disease, in particular a solid tumor, comprising a container a and a container B, wherein the container a contains an injectable aqueous solution, wherein the aqueous solution comprises a desired amount of an amino acid based compound and optionally other water soluble components according to formulation or administration concentration; and the container B contains an injectable oil solution, wherein the oil solution comprises a desired amount of lipids and optionally other lipophilic components according to formulation or administration concentration; wherein the ratio of the aqueous solution to the oil solution is such that in the emulsion thus formed, the concentration of the oil is < 50% by weight, and the weight ratio of the oil to the amino acid-based compound is not more than 8/1 or (1-320)/40, preferably not more than 4/1 or (1-160)/40. The definitions and descriptions of terms in this disclosure also apply to this device.

Based on the studies described in more detail below, the pharmaceutical composition of the present invention shows better performance in terms of emulsification, thereby allowing more effective effects of treatment using a drug comprising the pharmaceutical composition of the present invention, although specific mechanisms remain to be further studied.

Examples

The present invention is further illustrated by the following specific examples, which are not to be construed as limiting the invention thereto. In the following examples, all experimental animals were performed according to the relevant regulations and industry discipline. Unless otherwise specified, all tests were carried out according to the usual methods.

Materials, reagents and the like used in the following examples are commercially available unless otherwise specified. Some of the reagents used in the following examples are listed in Table 1.

TABLE 1

The experimental animals used in the following examples were all purchased from professional laboratory animals company and were all SPF (Specific Pathogen Free) grade animals. Taking mice as an example, there are 3 types: BALB/C mouse, nude mouse, C57BL/6 mouse, wherein the nude mouse is mutant line (BALB/C-nu) mouse obtained by introducing nude gene (nu) into BALB/C mouse. The mice are healthy females with the age of 6-8 weeks and the body weight of 17.5-20.5 g.

In the following examples, unless otherwise indicated, subcutaneous transplantation of tumor animals was performed according to the general practice of subcutaneous inoculation of solid tumor cells according to the guidelines issued by the drug administration. Unless otherwise indicated, solid tumors grow to the desired volume (e.g., mice carry tumors 30-500 mm)³) Then for successful modeling, the model was randomly divided into experimental groups of 6 animals each using PEMS 3.2 software (compiled by the national institutes of public health, western, university, Sichuan). Items for experimental observation, measurement and analysis include general state, body weight, food intake, animal graft versus host disease, solid tumor volume, tumor weight, survival time, and the like.

The tumor volume calculation formula is as follows:

tumor volume (V) ═ l/2 × a × b²Wherein a represents the tumor length and b represents the tumor width.

The tumor growth inhibition rate (abbreviated as tumor inhibition rate in the invention) is calculated by the following formula:

tumor inhibition rate Y (%) ═ (CW-TW)/CW × 100%, where TW is the average tumor weight of the study group; CW is the average tumor weight of the negative control group.

In the following examples, experimental results (e.g. tumor weights) are expressed as means ± standard deviation (x ± s), differences between two experimental animal groups and group means are compared by significance test using statistical software SPSS 13.0 or SPSS 19.0, tests are performed using statistic t, test level α is 0.05, P <0.05 indicates that the difference is statistically significant, otherwise it is not statistically significant.

Example 1: preparation of aqueous emulsions

Many different aqueous emulsions of the present invention can be formulated according to the above-described method of preparing the aqueous emulsion of the present invention. The composition of a portion of the aqueous emulsion of the present invention prepared in this example is shown in Table 2.

TABLE 2

Serial number	Oil and fat	Amino acid compound	Water (W)
				1	5% iodized oil	2.5% arginine/2.5% serine	90% water
2	1% iodized oil/5% ethyl oleate	5% arginine/5% serine	84% of water
				3	1% iodized oil/5% castor oil	5% arginine/5% serine	84% of water
4	25% iodized oil	15% arginine/15% serine	45% water

Several examples of the preparation tests of the aqueous emulsion of the invention are listed below.

For example, the amino acids (such as 2.5g arginine, 2.5g serine), oil (such as 5g super iodized oil), and 90ml water in the pharmaceutical composition are measured according to the required concentration. Then, the amino acid compound and water are fully mixed to form an aqueous solution, and the aqueous solution and the grease are respectively divided into required amounts (such as 19ml of the aqueous solution/bottle and 1ml of iodized oil/bottle) for storage and standby. In use, the aqueous amino acid compound solution is mixed well with the oil or fat in the desired amount ratio (e.g., 9.5: 0.5) and the mixture (e.g., preparation 1 in the above table) is ready for use as a contrast or/and embolization injection. Preparations 2-4 in the above table can also be prepared according to this method.

Example 2: preparation of water-based emulsion composition containing antitumor agent

According to the above-described method for preparing the antitumor-drug-containing pharmaceutical composition of the present invention, a wide variety of antitumor-drug-containing water-based emulsion compositions of the present invention can be formulated. The composition of a portion of the aqueous emulsion compositions of the present invention prepared in this example is shown in Table 3.

TABLE 3

Several examples of the preparation test of the water-based emulsion containing an antitumor agent of the present invention are listed below.

For example, amino acids (such as 5g arginine, 5g serine), oils (such as 7.5g polyoxyethylene 35-castor oil), 67.4ml water, and antitumor agent and other components (such as 10g ethanol, 5g propylene glycol, 0.1g taxol) are measured in the pharmaceutical composition according to the desired concentration. Then, the lipophilic component (e.g. taxol) is dissolved in ethanol/propylene glycol and polyoxyethylene 35-castor oil is added to form an oil solution, the amino acid compound, the hydrophilic component and water are fully mixed to form an aqueous solution, and the aqueous solution and the oil solution are respectively divided into required amounts (e.g. 7.74g of the aqueous solution/bottle and 2.26g of the oil solution/bottle) to be stored for later use. In use, the aqueous solution is mixed well with the oil solution in the desired amount ratio (e.g., 7.74: 2.26) and the mixture (e.g., preparation 1 in the above table) or a dilution of the mixture in water is ready for use as an intravenous injection. Preparation 2 in the above table was also prepared in this way.

For example, the amino acids (e.g. 20g arginine), lipids (e.g. 2.5g iodized oil, 12.5g ethyl oleate), 64.5ml water, and anti-tumor drugs and other ingredients (e.g. 0.5g docetaxel) in the pharmaceutical composition are measured according to the required concentration. Then, firstly, the lipophilic component (such as docetaxel) is dissolved in ethyl oleate and added with iodized oil to form an oil solution, then the amino acid compound, the hydrophilic component and water are fully mixed to form an aqueous solution, and the aqueous solution and the oil solution are respectively subpackaged into required amounts (such as 8.45 g/bottle of the aqueous solution and 1.55 g/bottle of the oil solution) for storage and standby. In use, the aqueous solution is mixed well with the oil solution in the desired amount ratio (e.g., 8.45: 1.55) and the mixture (e.g., preparation 3 in the above table) is ready for use as an intratumoral injection. Preparation 4 in the above table was also prepared in this way.

For example, amino acids (such as 20g arginine), oil (such as 5g super iodized oil), 74ml water, antitumor agent and other components (such as 1g 5-fluorouracil) in the pharmaceutical composition are measured according to the required concentration. Then, the amino acid compound, the water-soluble component (e.g., 5-fluorouracil) and water are fully mixed to obtain an aqueous solution, and the aqueous solution and the oil and fat are separately dispensed into required amounts (e.g., 9.5g of the aqueous solution per bottle and 0.5g of iodized oil per bottle) and stored for later use. In use, the aqueous solution is mixed well with the oil or fat in a desired amount ratio (e.g., 9.5: 0.5) and the mixture (e.g., preparation 5 of the above table) is used as an intratumoral injection. Preparations 6-13 in the above table were also prepared in this way.

For example, the water solution packaged in the above preparations (for example, 9-9.5 g/bottle) is cooled to 10-25 deg.C, and then freeze-dried, corked and covered to prepare sterilized dry powder for use. When in use, the sterilized dry powder is dissolved in about corresponding amount of water for injection (such as 7.5-8 ml/bottle) (if difficult, the solution can be heated to 60 deg.C), and then corresponding amount of oil or fat solution is added into the solution and mixed for use.

Example 3: research on action of amino acid compound in oil-water emulsification

Emulsion samples and their compositions are shown in the table below, and were prepared as follows: firstly, respectively measuring and adding distilled water with variable weight according to the required concentration of samples 1-4 (the sample 1 is 4ml, the sample 2 is 9ml, and the samples 3 and 4 are 7ml) into 4 beakers, then placing the beakers to a magnetic stirrer for stirring (the rotating speed is 600rpm/min), respectively measuring and adding amino acid compounds with variable weight according to the required concentration of the samples 1-4 (the samples 1 and 2 are 0g, and the samples 3 and 4 are 2g respectively) to prepare an aqueous solution, and then respectively measuring and adding super-liquefied iodine oil with variable weight according to the required concentration of the samples 1-4 (the sample 1 is 6g, and the samples 2, 3 and 4 are 1g) into 4 beakers respectively. After stirring was continued for 5 minutes, 1ml of liquid was drawn in from the beaker using 4X 6 (6 in each case) 1ml syringes, each of which was observed for the time at which demixing occurred in the standing still state. The time to delamination for each sample is given in table 4.

TABLE 4

Sample number	Oil and fat concentration	Water concentration	Amino acid compound and concentration thereof	Time to layer (min)
					1	60％	40％	0	Are all made of>60
2	10％	90％	0	Are all made of<0.25
					3	10％	70％	20% arginine	50±3
4	10％	70％	10% arginine/10% tryptophan	48±4

Emulsions formed by mixing fats and oils with aqueous solutions exhibit separation of the oil and aqueous phases under quiescent conditions. The layering speed is an important index for measuring the emulsification quality and the stability thereof. In the above table, the difference in time between stratification for sample 2 and sample 1 is statistically significant (P < 0.05). In accordance with the teachings of the prior art, the ultra-liquefied iodized oil is well emulsified with water only at a relatively high concentration, which seems to be consistent with this result.

However, despite their same oil concentration (10%), the difference in the stratification times for samples 3, 4 and 2 were statistically significant (P <0.05, respectively), whereas the difference in the stratification times for samples 3 and 4 was not statistically significant (P >0.05), and the stability of the aqueous emulsion reflected by the stratification times could meet the clinical dosing requirements.

According to the above and more similar studies, the amino acids can improve the stability of the water-oil emulsion, or/and reduce the concentration of lipids required for the preparation of the emulsion (at least by < 50%, preferably < 30%). Thus, the amino acid compound can form the aqueous emulsion of the present invention together with a low-concentration oil or fat.

Example 4: research on amino acid compound/oil content ratio in water-based emulsion

Emulsion samples and their compositions are shown in the table below, and were prepared as follows: firstly, respectively measuring and measuring the distilled water with the variable weight (9 ml, 8.88ml, 8.75ml, 8.5ml, 8.0ml and 7.0ml) according to the required concentration of the sample 1-6, adding the distilled water into 6 beakers, then placing the beakers to a magnetic stirrer for stirring (rotating speed of 600rpm/min), respectively measuring and adding the arginine with the variable weight (0 g, 0.125g, 0.25g, 0.5g, 1.0g and 2.0g) according to the required concentration of the sample 1-6 for preparing the aqueous solution, and then respectively measuring and measuring 1g of the super-liquefied iodine oil and adding the super-liquefied iodine oil into 6 beakers. After stirring was continued for 5 minutes, 1ml of liquid was drawn in from the beaker using 6X 6 (6 per group) 1ml syringes each of which was observed in a standing still state for the time at which each layer appeared. The time to delamination for each sample is given in table 5.

TABLE 5

Sample number	Amount ratio of oil/amino acid Compound	Oil and fat concentration	Amino acid compound concentration	Time to layer (min)
					1	-	10％	0	Are all made of<1
2	10/1.25	10％	1.25％	25±2
					3	10/2.5	10％	2.5％	30±8
4	10/5	10％	5％	36±4
					5	10/10	10％	10％	48±7
6	10/20	10％	20％	52±3

In the above table, the delamination occurred almost instantaneously in sample 1, while the delamination time differences between sample 2 and sample 1 were statistically significant (P <0.05), the delamination time differences between samples 3 to 6 and sample 1, respectively, were also statistically significant (P <0.05), and the delamination time of the aqueous emulsion increased with the increase in the amount ratio of the oil/amino acid compound.

According to the above-mentioned studies and more similar studies, amino acid compounds are essential components in the aqueous emulsion of the present invention, provided that: the amount ratio (w/w) of the oil/amino acid compound is not more than 8/1 or (1-320)/40, preferably not more than 4/1 or (1-160)/40.

Example 5: selection of amino acid compounds

Emulsion samples and their compositions are shown in the table below, all formulated as in example 1. Firstly, respectively measuring 10g to 6 beakers of amino acid compounds shown in the following table, respectively adding water, placing the mixture on a magnetic stirrer to stir (rotating speed is 600rpm/min) to prepare 9.5ml of aqueous solution, respectively measuring the super-liquefied iodized oil with the required weight (0.5 g/beaker), respectively adding the super-liquefied iodized oil into each beaker, and mixing for 5 minutes. Then, 1ml of the liquid was drawn from the beaker by using 6X 6 (6 in each group) 1ml syringes, respectively, and the time at which each delamination occurred was observed in the standing still state of the syringes. The time to delamination for each sample is given in table 6.

TABLE 6

Sample number	Oil and fat concentration	Amino acid compound and concentration thereof	Time to layer (min)
				1	10％	10% lysine	<5
2	10％	10% proline	<5
				3	10％	10% valine	<5
4	10％	10% alanine	<5
				5	10％	10% arginine	51±4
6	10％	5% arginine/5% serine	49±6

In the above table, samples 5 and 6 are clearly distinguished from the other samples. Surprisingly, arginine has a high similarity to lysine, but the emulsion stability of the latter is much lower than that of the former.

Example 5: emulsion study of pharmaceutical composition comprising antitumor drug

Emulsion samples and their compositions are shown in the table below, and were prepared as follows: firstly, distilled water with different weights (sample 1 is 7ml, samples 2 and 5 are 6.8ml, samples 3, 6 and 7 are 6.9ml, sample 4 is 6.99ml, sample 8 is 6ml and sample 9 is 5.9ml) is respectively measured according to the required concentration of samples 1-9 and added into 9 beakers, and then respectively placing the mixture in a magnetic stirrer for stirring (the rotating speed is 600rpm/min), respectively adding the same amount of amino acid compounds (samples 1-9 are 1g of arginine and 1g of serine) into cups for dissolving, respectively measuring and adding the antitumor drugs (sample 1 is 0, samples 2 and 5 are 0.2g, samples 3, 6 and 7 are 0.1g, sample 4 is 0.01g, sample 8 is 1g and sample 9 is 1.1g) according to the required concentration of the samples 1-9 for preparing aqueous solution, and respectively adding the same amount of super-liquefied iodine oil (samples 1-9 are 1g) into 9 beakers. After stirring was continued for 5 minutes, 1ml of the liquid was drawn in from the beaker using 9X 6 (6 in each case) 1ml syringes, which were observed in the standing still state for the time at which each layer had appeared. The time to delamination for each sample is given in table 7.

TABLE 7

In the above table, the emulsion samples to which the above antitumor agents were added all showed stability consistent with that of sample 1 even in the group to which a larger amount of antitumor agent (samples 7, 8) was added.

According to the above-mentioned studies and more similar studies, the addition of an antitumor agent (conventional antitumor agent or/and non-conventional antitumor agent) at an effective injection concentration within the above-mentioned amount ratio of the oil/amino acid-based compound still forms a stable injectable aqueous emulsion. The following experiments further optimize the composition of the pharmaceutical composition emulsion containing the antitumor agent.

In one experiment, the experimental animals were BALB/c mice, and the modeled cells were breast cancer 4T1 cells at 1X 10⁶Individual cells/animal right axillary subcutaneous transplantation tumor modeling. Successfully modeled test animals (mean tumor volume 278 mm)³) The groups were randomly divided into 1 negative control group (0) and 6 drug study groups (1-6). The negative control was physiological saline and study drugs are shown in the table below. 6 drugs each contain 5-fluoroUracil/arginine aqueous solution and super iodized oil were prepared according to the preparation method of example 2. Specifically, 35ml of 1% 5-fluorouracil/20% arginine aqueous solution is prepared, 4.9ml of aqueous solution is measured and added into 6 beakers, the volume-variable water solution for injection is added according to the oil concentration required by the samples 1-6, the aqueous solution is pumped into a 1ml syringe for standby, and the required amount of super-liquefied iodized oil is mixed with the required amount of oil concentration required by the samples 1-6 to form the emulsion when the emulsion is used. The intratumoral injection is used as the intratumoral administration mode in each experimental group. Each group was administered once every 3 days for a total of 3 times, and the injection amount was 150. mu.l/tube. On day 3 after the end of the administration, the animals were euthanized, and the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group, and the results are shown in table 8.

TABLE 8

It is generally believed that the oil in the emulsion for intratumoral administration is also a sustained release or/and embolization vehicle, and that sustained release or/and embolization is beneficial for improving the efficacy of the drug. However, in the above table, group 7 using high concentration of lipids according to the prior art showed a tumor inhibition rate of < 40%. In fact, high concentrations of lipids at least limit the maximum concentration of water-soluble antineoplastic agents in the emulsion.

The above studies and more similar studies show that the concentration of oil is preferably 30% or less or 1 to 30%, more preferably 20% or 1 to 20%, or 10% or 1 to 10% in the composition emulsion comprising the antitumor agent. The following experiments have studied more tumors under this preferred condition.

In each of the following experiments, the successfully modeled human cancer cell-bearing nude mice were randomly divided into 1 negative control group and 6 study groups (A, B, C, D, E, F groups). The corresponding negative control was physiological saline, and the A, B, C, D, E, F groups used 6 study drugs in order: 20% arginine/1% 5-fluorouracil aqueous solution, 20% arginine/2.5% super-liquefied iodized oil/1% 5-fluorouracil/76.5% aqueous emulsion, 10% arginine/10% serine/7% sodium bicarbonate/3% sodium hydroxide aqueous solution, 10% arginine/10% serine/2.5% super-liquefied iodized oil/7% sodium bicarbonate/3% sodium hydroxide/67.5% aqueous emulsion, 5% arginine/5% glycine/1% methylene blue aqueous solution, 5% arginine/5% glycine/2.5% super-liquefied iodized oil/1% methylene blue/86.5% aqueous emulsion. Wherein: the aqueous solution is prepared according to a conventional aqueous solution preparation method (solid or/and liquid antitumor drugs are dissolved in water), and the emulsion is prepared according to the preparation method of example 2 (aqueous solution and super-liquefied iodized oil are mixed and emulsified). Each group was injected intratumorally, once every 3 days for a total of 3 times, 150. mu.l/patient. On day 3 after the end of the administration, the animals were euthanized, and the tumor weight was measured after dissection, and the tumor inhibition rate was calculated from the negative control group.

1) Application of the compound in pancreatic tumor treatment

In the study, the nude mouse with human pancreatic cancer cells (PANC-1) (average tumor volume 302 mm) successfully modeled³) The groups were randomized into a negative control group and 6 study groups (A, B, C, D, E, F groups). A. The tumor inhibition rates of B, C, D, E, F groups were: 95.6 percent, 91.7 percent, 97.3 percent, 94.6 percent, 91.2 percent and 92.3 percent, which all meet the commonly considered effective anti-tumor standard (the tumor inhibition rate is more than or equal to 40 percent).

2) Application of the compound in lung tumor treatment

In the study, the nude mouse with human lung cancer cells (A549) successfully modeled (mean tumor volume 249 mm)³) The groups were randomized into a negative control group and 6 study groups (A, B, C, D, E, F groups). A. The tumor inhibition rates of B, C, D, E, F groups were: 86.2 percent, 81.3 percent, 91.3 percent, 89.6 percent, 87.4 percent and 85.3 percent, which all meet the commonly considered effective anti-tumor standard (the tumor inhibition rate is more than or equal to 40 percent).

3) Application of the compound in thyroid tumor treatment

In the study, the nude mice bearing human thyroid cancer cells (SW579) successfully modeled (mean tumor volume 261 mm)³) The groups were randomized into a negative control group and 6 study groups (A, B, C, D, E, F groups). A. The tumor inhibition rates of B, C, D, E, F groups were: 83.1%, 76.4%, 88.1%, 87.5%, 89.6%, 88.4%, all of which are satisfiedThe standard of effective anti-tumor (tumor inhibition rate is more than or equal to 40 percent) is generally considered.

4) Use in the treatment of prostate tumors

In this study, human prostate cancer cell (LNCaP/AR) -bearing nude mice (mean tumor volume 273 mm) were successfully modeled³) The groups were randomized into a negative control group and 6 study groups (A, B, C, D, E, F groups). A. The tumor inhibition rates of B, C, D, E, F groups were: 82.7%, 79.1%, 92.4%, 91.3%, 89.1%, 86.9% all meet the commonly recognized effective anti-tumor standard (the tumor inhibition rate is more than or equal to 40%).

5) Application of the compound in liver tumor treatment

In the study, the nude mice (average tumor volume 297 mm) with human hepatoma cells (HepG2) successfully modeled³) The groups were randomized into a negative control group and 6 study groups (A, B, C, D, E, F groups). A. The tumor inhibition rates of B, C, D, E, F groups were: 94.3 percent, 90.2 percent, 93.5 percent, 89.3 percent, 92.4 percent and 91.3 percent, which all meet the commonly considered effective anti-tumor standard (the tumor inhibition rate is more than or equal to 40 percent).

6) Application of the compound in treating head and neck tumors

In this study, nude mice with human head and neck cancer cells (F μ da) successfully modeled (average tumor volume 267 mm)³) The groups were randomized into a negative control group and 6 study groups (A, B, C, D, E, F groups). A. The tumor inhibition rates of B, C, D, E, F groups were: 83.1%, 81.6%, 91.4%, 92.7%, 90.5%, 89.2%, all meet the commonly recognized effective anti-tumor standard (tumor inhibition rate is more than or equal to 40%).

7) Application of the compound in treatment of nasopharyngeal tumors

In the study, nude mice (mean tumor volume 258 mm) with human nasopharyngeal carcinoma cells (CNE1) successfully modeled³) The groups were randomized into a negative control group and 6 study groups (A, B, C, D, E, F groups). A. The tumor inhibition rates of B, C, D, E, F groups were: 81.4%, 79.1%, 90.2%, 86.4%, 93.5%, 88.4%, all meet the commonly recognized effective anti-tumor standard (tumor inhibition rate is more than or equal to 40%).

8) Application of the compound in treating gastric tumor

In the research experiment, the nude mouse with human gastric cancer cell (BGC823) successfully modeled (average tumor volume 264 mm)³) The groups were randomized into a negative control group and 6 study groups (A, B, C, D, E, F groups). A. The tumor inhibition rates of B, C, D, E, F groups were: 81.4%, 79.1%, 90.2%, 86.4%, 93.5%, 88.4%, all meet the commonly recognized effective anti-tumor standard (tumor inhibition rate is more than or equal to 40%).

9) Application of the compound in ovarian tumor treatment

In the study, nude mice (average tumor volume 291 mm) with human ovarian carcinoma cells (PA1) successfully modeled³) The groups were randomized into a negative control group and 6 study groups (A, B, C, D, E, F groups). A. The tumor inhibition rates of B, C, D, E, F groups were: 79.6 percent, 78.3 percent, 89.3 percent, 84.6 percent, 91.1 percent and 83.9 percent meet the commonly considered effective anti-tumor standard (the tumor inhibition rate is more than or equal to 40 percent).

Similar results were obtained with some other compositions of the invention prepared by the method of example 2, for use in the treatment of each of the tumors described above.

The present disclosure relates to the following items:

1. a stable water-based emulsion comprising an amino acid-based compound and an oil or fat, and in which emulsion the concentration of the oil or fat is < 50% by weight, and the weight ratio of the oil or fat to the amino acid-based compound is not more than 8/1 or (1-320)/40, preferably not more than 4/1 or (1-160)/40.

2. The emulsion according to item 1, wherein the concentration (w/w) of the oil or fat in the emulsion is 30% or less or 1 to 30%, or 20% or 1 to 20%, or 10% or less or 1 to 10%.

3. The emulsion according to item 1 or 2, wherein the oil or fat is selected from an oil for injection and a derivative thereof, wherein the oil for injection is one or more selected from the group consisting of: ethyl oleate, medium-chain (C6-10) fatty acid glyceride, isopropyl myristate, soybean oil, tea oil, olive oil, castor oil, peanut oil, cottonseed oil, corn oil, linseed oil, poppy seed oil and sesame oil.

4. The emulsion according to clause 1 or 2, wherein the oil comprises an iodized oil.

5. The emulsion according to item 1 or 2, wherein the amino acid-based compound is selected from one or more of the following amino acids and derivatives thereof: arginine, alanine, glutamic acid, glycine, histidine, lysine, valine, isoleucine, leucine, phenylalanine, threonine, proline, serine.

6. The emulsion according to item 5, wherein the amino acid based compound comprises arginine.

7. Use of an aqueous emulsion according to one of items 1 to 6 for the preparation of a pharmaceutical composition for the treatment of solid tumors.

8. The use according to clause 7, wherein the pharmaceutical composition further optionally comprises one or more antineoplastic agents.

9. A pharmaceutical composition for the treatment of solid tumors comprising an aqueous-based emulsion according to one of items 1 to 6.

10. The pharmaceutical composition according to item 9, further comprising one or more antineoplastic agents.

11. A method of treating a solid tumor comprising administering in vivo to an individual in need thereof a therapeutically effective volume of an aqueous-based emulsion according to one of items 1-6 or a pharmaceutical composition according to item 9 or 10.

12. The use, pharmaceutical composition or method according to one of items 1 to 11, wherein the concentration (w/w) of the amino acid(s) in the pharmaceutical composition is > 3%, or ≧ 5%, or 5-50%.

13. The use, pharmaceutical composition or method according to one of items 1 to 12, wherein the concentration (w/w) of the lipid in the pharmaceutical composition is 15% or less or 0.5 to 15%, more preferably 15% or 0.5 to 15%, or 5% or less or 0.5 to 5%.

14. The use or pharmaceutical composition according to item 8 or 10, wherein the antineoplastic agent is selected from one or more of an atypical antineoplastic agent or/and a conventional antineoplastic agent.

15. The use or pharmaceutical composition according to item 14, wherein in said pharmaceutical composition, if present, the concentration (w/w) of said non-conventional anti-neoplastic drug is more than 0.25%, preferably 0.35-40%; if present, the conventional antineoplastic agent is present at a concentration (w/w) of greater than 0.05%, 0.05-15%, preferably 0.5-10%.

16. The use or pharmaceutical composition according to item 14 or 15, wherein the conventional anti-neoplastic drug comprises one or more selected from the group consisting of: drugs that disrupt the structure and function of DNA, drugs that interfere with the transcription of RNA that intercalate into DNA, drugs that interfere with DNA synthesis, drugs that affect protein synthesis.

17. The use or pharmaceutical composition according to item 14 or 15, wherein the non-conventional anti-neoplastic drug is selected from one or more of the group consisting of: vital dye in a concentration (w/w) of 0.5-15% in the pharmaceutical composition; a strong base at a concentration of 0.5% or more, or 0.5-6%, preferably 0.5-1.5% or 1.5-6% in the pharmaceutical composition; a weak base at a concentration (w/w) in the pharmaceutical composition of 3.5-50%; a strong acid at a concentration of 0.5% or more, or 0.5-3%, preferably 0.5-1.5% or 1.5-3% in the pharmaceutical composition; a weak acid in a pharmaceutical composition at a concentration (w/w) of 0.5-50%.

18. The use or pharmaceutical composition according to one of items 14 to 17, wherein the antineoplastic agent comprises the conventional antineoplastic agent and the vital dye.

19. The use or pharmaceutical composition according to clauses 17 or 18, wherein the vital dye is one or more selected from the group consisting of: methylene blue, patent blue, isothio blue, bengal red, and the concentration (w/w) of the vital dye in the pharmaceutical composition is 0.5% or more, 0.5-15%, preferably 0.5-1.5% or 3.5-15%.

20. The use or pharmaceutical composition according to one of items 14 to 17, wherein the pharmaceutical composition comprises the conventional anti-neoplastic drug and a non-conventional anti-neoplastic drug selected from one or more of the following: the strong base, the weak base, the strong acid, the weak acid.

21. The use or pharmaceutical composition according to clauses 17 or 20, wherein the strong base is sodium hydroxide.

22. The use or pharmaceutical composition according to clauses 17 or 20, wherein the weak base is one or more selected from the group consisting of: sodium phosphate, sodium carbonate, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium bicarbonate, potassium hydrogen phthalate, sodium acetate, sodium propionate, sodium butyrate, sodium malonate, sodium lactate, sodium citrate, sodium malate and sodium dodecyl sulfate.

23. The use or pharmaceutical composition according to clauses 17 or 20, wherein the strong acid is hydrochloric acid.

24. The use or pharmaceutical composition according to clause 17 or 20, wherein the weak acid is one or more selected from the group consisting of: acetic acid, propionic acid, butyric acid, malonic acid, lactic acid, citric acid, malic acid.

25. The use or pharmaceutical composition according to one of items 14 to 24, wherein the conventional anti-neoplastic drug is one or more selected from the group consisting of: uracil derivatives, cyclophosphamide, gemcitabine, epirubicin, antitumor antibiotics, teniposide, metal platinum complex and taxanes.

26. The use or pharmaceutical composition according to one of items 14 to 25, wherein the conventional antitumor agent is one or more selected from the group consisting of: doxorubicin hydrochloride, gemcitabine, cisplatin, 5-fluorouracil, alexancept, carboplatin, mitomycin, captopop, cyclophosphamide, doxorubicin, carmustine, camptothecin, paclitaxel, vincristine, elemene, docetaxel, fotemustine.

27. The use, pharmaceutical composition or method according to items 2-7, wherein said tumor comprises a malignant tumor and a non-malignant tumor.

28. The use, pharmaceutical composition for use or method according to item 27, wherein the malignant tumor comprises breast cancer, pancreatic cancer, thyroid cancer, nasopharyngeal cancer, prostate cancer, liver cancer, lung cancer, intestinal cancer, oral cancer, esophageal cancer, gastric cancer, laryngeal cancer, testicular cancer, vaginal cancer, uterine cancer, ovarian cancer.

29. The use, pharmaceutical composition or method according to one of items 6 to 28, wherein the method comprises optionally further performing one or more other treatments, such as chemotherapy, immunotherapy, radiotherapy, surgery, physical ablation, before, during or after administration of the pharmaceutical composition.

30. A method for preparing an aqueous emulsion according to one of items 1 to 6, comprising the steps of: 1) preparing an aqueous solution comprising the amino acid compound and optionally other water-soluble components; 2) preparing an oil solution comprising lipids and optionally other lipophilic components; and 3) sufficiently mixing the aqueous solution prepared in the step 1) and the oil solution prepared in the step 2) into an injectable emulsion.

31. A method of preparing a pharmaceutical composition according to clause 9 or 10, comprising the steps of: 1) preparing an aqueous solution comprising the amino acid-based compound, optionally a water-soluble antineoplastic agent and optionally other water-soluble components; 2) preparing an oil solution comprising lipids, optionally lipophilic antineoplastic agents and optionally other lipophilic components; 3) mixing the aqueous solution prepared in step 1) with the oil solution prepared in step 2) thoroughly.

32. A device for treating a solid tumor comprising a container a and a container B, wherein the container a contains an injectable aqueous solution, wherein the aqueous solution comprises the desired amount of an amino acid based compound and optionally other water soluble components according to formulation or dosing concentration; and the container B contains an injectable oil solution, wherein the oil solution comprises a desired amount of lipids and optionally other lipophilic components according to formulation or administration concentration; wherein the ratio of the aqueous solution to the oil solution is such that in the emulsion thus formed, the concentration of the oil is < 50% by weight, and the weight ratio of the oil to the amino acid-based compound is not more than 8/1 or (1-320)/40, preferably not more than 4/1 or (1-160)/40.

33. The emulsion according to item 4, which is used as a developer.

34. A developer which is an aqueous based emulsion comprising an iodized oil and an amino acid based compound, wherein in the emulsion the concentration of said iodized oil is < 50% by weight and the weight ratio of said iodized oil to said amino acid based compound is < 8/1 or (1-320)/40, preferably < 4/1 or (1-160)/40.

Various modifications of the invention in addition to those described herein will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference, including all patents, patent applications, journal articles, books, and any other publications, cited in this application is hereby incorporated by reference in its entirety.

Claims

2. An emulsion according to claim 1, wherein the concentration (w/w) of said oil or fat in the emulsion is 30% or less or 1-30%, or 20% or 1-20%, or 10% or 1-10%.

3. An emulsion according to claim 1 or 2 wherein the oil or fat is selected from an oil for injection and derivatives thereof, wherein the oil for injection is one or more selected from: ethyl oleate, medium-chain (C6-10) fatty acid glyceride, isopropyl myristate, soybean oil, tea oil, olive oil, castor oil, peanut oil, cottonseed oil, corn oil, linseed oil, poppy seed oil and sesame oil.

4. An emulsion according to claim 1 or 2, wherein the amino acid compound is selected from one or more of the following amino acids and derivatives thereof: arginine, alanine, glutamic acid, glycine, histidine, lysine, valine, isoleucine, leucine, phenylalanine, threonine, proline, serine.

5. Use of an aqueous emulsion according to one of claims 1 to 4 for the preparation of a pharmaceutical composition for the treatment of solid tumors.

6. A pharmaceutical composition for the treatment of solid tumors comprising the aqueous-based emulsion according to any one of claims 1-4.

7. The pharmaceutical composition according to claim 6, further comprising one or more antineoplastic agents.

8. Process for the preparation of an aqueous emulsion according to one of claims 1 to 4, comprising the following steps: 1) preparing an aqueous solution comprising the amino acid compound and optionally other water-soluble components; 2) preparing an oil solution comprising lipids and optionally other lipophilic components; and 3) sufficiently mixing the aqueous solution prepared in the step 1) and the oil solution prepared in the step 2) into an injectable emulsion.

9. A device for treating a solid tumor comprising a container a and a container B, wherein the container a contains an injectable aqueous solution, wherein the aqueous solution comprises the desired amount of an amino acid based compound and optionally other water soluble components according to formulation or dosing concentration; and the container B contains an injectable oil solution, wherein the oil solution comprises a desired amount of lipids and optionally other lipophilic components according to formulation or administration concentration; wherein the ratio of the aqueous solution to the oil solution is such that in the emulsion thus formed, the concentration of the oil is < 50% by weight, and the weight ratio of the oil to the amino acid-based compound is not more than 8/1 or (1-320)/40, preferably not more than 4/1 or (1-160)/40.

10. A developer which is an aqueous based emulsion comprising an iodized oil and an amino acid based compound, wherein in the emulsion the concentration of said iodized oil is < 50% by weight and the weight ratio of said iodized oil to said amino acid based compound is < 8/1 or (1-320)/40, preferably < 4/1 or (1-160)/40.