Pharmaceutical composition for treating acute kidney injury and application thereof

Abstract

The invention specifically discloses a pharmaceutical composition for treating acute kidney injury and application thereof, and relates to the technical field of biological medicines. The invention provides a pharmaceutical composition. The pharmaceutical composition comprises: (1) a therapeutically effective amount of sodium citrate and methionine; and (2) a pharmaceutically or immunologically acceptable carrier or excipient. The pharmaceutical composition provided by the invention can effectively prevent acute kidney injury through an intraperitoneal injection way, and provides a new treatment way for acute kidney injury related to rhabdomyolysis.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to a pharmaceutical composition for treating acute kidney injury and application thereof. Background Art

[0002] Acute kidney injury (AKI) is a life-threatening condition. In the past few years, the application of uniform definitions and classifications has shown that even mild forms of AKI, documented by a minimal and fully reversible increase in plasma creatinine, are associated with reduced patient survival and progression of chronic kidney disease. AKI affects patients from all medical disciplines, for example during sepsis or after surgery, with ischemia characterized by lack of oxygen and nutrient supply being its main cause. It occurs particularly in critically ill patients, for example after cardiac surgery and liver transplantation, with an incidence of up to 50%.

[0003] Citrate shows protective effects on cardiovascular and renal function in ischemia-induced acute kidney injury. BMC Nephrol 18, 130 (2017). It mentioned that an acute kidney injury model was established in rats by ischemia-reperfusion (I / R), and different concentrations of sodium citrate (0.3mmol / kg / h; 0.6mmol / kg / h and 1.0mmol / kg / h) were continuously infused before and 45 minutes after ischemia. It was found that compared with the I / R control group, the infusion of 1.0mmol / kg / h of citrate led to a decrease in creatinine and LDH activity, and an increase in tissue ATP content, indicating that citrate has a protective effect during I / R-induced AKI.

[0004] However, this sodium citrate preparation only exerts a protective effect when administered before kidney damage. When administered after damage, the protective effect is limited, which restricts the use of this preparation after AKI occurs. In addition, the pH of the sodium citrate solution of 1.0 mmol / kg / h is low, which is far from the normal physiological pH value, and intravenous use increases vascular irritation. On the other hand, the sodium citrate used is in a high concentration. In kidney damage, due to the decrease in metabolism, citric acid accumulation and acidosis may occur, resulting in more severe systemic symptoms caused by the kidney. Summary of the invention

[0005] 1. Technical issues to be resolved

[0006] In view of this, one of the main purposes of the present invention is to use sodium citrate and methionine in the preparation of a medicament for preventing and / or treating acute kidney injury and / or diseases and / or symptoms associated with acute kidney injury.

[0007] (II) Technical solution

[0008] In order to achieve the above object, the present invention provides a pharmaceutical composition, which comprises:

[0009] (1) a therapeutically effective amount of sodium citrate and methionine;

[0010] (2) A pharmaceutically or immunologically acceptable carrier or excipient.

[0011] In one embodiment, the pharmaceutical composition further comprises citric acid and / or sodium chloride.

[0012] In one embodiment, the mass ratio of sodium citrate to methionine is 3.08:2.0-4.3:2.0.

[0013] In one embodiment, the amount of sodium citrate used is 4.3 g; and / or the amount of methionine used is 2.0 g.

[0014] In one embodiment, the mass ratio of sodium citrate, methionine, citric acid and sodium chloride is 3.08:2.0:1.12:9.0-4.3:2.0:1.2:9.0 。

[0015] In one embodiment, the mass ratio of sodium citrate, methionine, citric acid and sodium chloride is 4.3:2.0:1.2:9.0.

[0016] In one embodiment, the amount of sodium citrate is 4.3 g; and / or the amount of methionine is 2.0 g; and / or the amount of citric acid is 1.2 g; and / or the amount of sodium chloride is 9.0 g.

[0017] In another aspect, the present invention further provides a pharmaceutical preparation, which comprises the above-mentioned pharmaceutical composition.

[0018] In one embodiment, the administration route of the pharmaceutical preparation comprises one or a combination of intranasal, intrathecal, intraarterial, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, ocular, sublingual, oral, intracerebral and transdermal.

[0019] In one embodiment, the route of administration is intraperitoneal injection.

[0020] In another aspect, the present invention also provides use of the above-mentioned pharmaceutical composition or pharmaceutical preparation in the preparation of a medicament for preventing and / or treating acute kidney injury and / or diseases and / or symptoms associated with acute kidney injury.

[0021] In one embodiment, said acute kidney injury is induced by rhabdomyolysis.

[0022] In one embodiment, the pharmaceutical composition or pharmaceutical preparation is administered 0-3 hours after rhabdomyolysis-induced acute kidney injury model.

[0023] In one embodiment, the pharmaceutical composition or pharmaceutical preparation is administered at 0 h after rhabdomyolysis-induced acute kidney injury model.

[0024] In one embodiment, the dosage of the pharmaceutical composition or pharmaceutical preparation is 0-400ul / mouse.

[0025] In one embodiment, the dosage is 200ul / mouse.

[0026] In one embodiment, the administering subject is healthy.

[0027] In one embodiment, the administering subject is non-healthy.

[0028] In one embodiment, the subject of administration suffers from acute kidney injury.

[0029] In one embodiment, the administering subject does not suffer from acute kidney injury.

[0030] In one embodiment, the subject includes mammals and non-mammals.

[0031] Examples of mammals include, but are not limited to, any member of the class Mammalia: humans, non-human primates such as chimpanzees and other apes and monkeys; farm animals such as cattle, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs and cats; laboratory animals, including rodents such as rats, mice and guinea pigs, etc. Examples of non-mammals include, but are not limited to, birds, fish or other non-mammals, etc.

[0032] In one embodiment, the subject is a mouse.

[0033] (III) Beneficial effects

[0034] The present invention provides the use of sodium citrate and methionine in the preparation of a medicament for preventing and / or treating acute kidney injury and / or diseases and / or symptoms associated with acute kidney injury. Compared with the prior art, the medicament has the following beneficial effects:

[0035] 1. Intraperitoneal injection is simpler and can be done as a single injection or multiple injections over multiple days.

[0036] 2. The solution has stable properties, close to the natural pH value and osmotic pressure of the human body, which can reduce the irritation of the preparation to local tissues.

[0037] 3. There are obvious advantages in improving the degree of kidney damage, with indicators such as creatinine and urea nitrogen decreasing by more than 30%.

[0038] 4. Currently, the main treatment options for acute kidney injury caused by rhabdomyolysis include eliminating the cause of rhabdomyolysis, hydration (mainly intravenous infusion of normal saline), alkalinizing urine, supplementing vitamins and proteins to prevent further muscle dissolution, and initiating renal dialysis treatment once acute renal failure occurs. The drug combination provided by the present invention can effectively prevent the occurrence of acute kidney injury through intraperitoneal injection, and provides a new treatment approach for acute kidney injury related to rhabdomyolysis.

[0039] (IV) Terms and Definitions

[0040] As used herein, the terms "patient" or "subject" are used interchangeably and refer to mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class Mammalia: humans, non-human primates such as chimpanzees and other apes and monkeys; farm animals such as cattle, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs and cats; laboratory animals, including rodents such as rats, mice and guinea pigs, etc. Examples of non-mammals include, but are not limited to, birds, fish or other non-mammals, etc.

[0041] As used herein, the term "pharmaceutical composition" refers to a composition comprising an immune checkpoint inhibitor and / or a histone acetyltransferase inhibitor formulated together with one or more pharmaceutically acceptable carriers.

[0042] The formulation of the pharmaceutical composition can be adjusted according to the application. In particular, the pharmaceutical composition can be formulated using methods known in the art to provide quick, continuous or delayed release of the active ingredient after administration to a mammal.

[0043] The dosage form of the pharmaceutical composition disclosed in the present invention can be granules, tablets, lyophilized powder, suppositories, capsules, sublingual tablets, liquid solutions, nasal drops, sprays, and quantitative sprays.

[0044] The pharmaceutical compositions of the present invention may be administered using any known method.The term "administering" or "applying" a substance, compound or agent to a subject may be performed using one of a variety of methods known to those skilled in the art.

[0045] For example, the compound or agent can be administered intranasally (e.g., by inhalation), intrathecally (into the spinal canal or subarachnoid space), intraarterially, intradermally, intramuscularly, intraperitoneally, intravenously, subcutaneously, ocularly, sublingually, orally (by ingestion), intracerebrally, and transdermally (by absorption, e.g., through a skin catheter). The compound or agent can also be suitably introduced by a rechargeable or biodegradable polymeric device or other device (e.g., a patch and a pump or formulation) that provides extended, slowed, or controlled release of the compound or agent. Administration can also be performed, for example, once, multiple times, and / or over one or more extended periods of time.

[0046] As used herein, the term "pharmaceutically acceptable" ingredients are substances that are suitable for use in humans and / or animals without excessive adverse reactions (such as toxicity, irritation, and allergic response), ie, at a reasonable benefit / risk ratio.

[0047] As used herein, the term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" is a carrier for the administration of therapeutic agents, including various excipients and diluents. The term refers to such pharmaceutical carriers: they are not necessary active ingredients themselves and are not overly toxic after administration. Suitable carriers are well known to those of ordinary skill in the art, and a full discussion of pharmaceutically acceptable excipients can be found in Remington's Pharmaceutical Sciences (Mack Pub. Co., NJ 1991).

[0048] In the composition, pharmaceutically acceptable carriers include any and all solvents, dispersion media, preservatives, antioxidants, coatings, isotonic and absorption delaying agents, surfactants, fillers, disintegrants, adhesives, diluents, lubricants, glidants, pH regulators, buffers, enhancers, wetting agents, solubilizers, surfactants, antioxidants, etc. that are compatible with drug administration. The use of such media and agents for pharmaceutically active substances is well known in the art. The composition may contain other active compounds that provide supplementary, additional or enhanced therapeutic functions. Solid carriers or excipients, such as lactose, starch or talc, or liquid carriers, such as water, fatty oils or liquid paraffin. Other examples of the carrier include culture media, such as DM EM or RPMI; and low temperature storage media, comprising components that scavenge free radicals, provide pH buffering, osmotic / osmotic support, energy substrates and ion concentrations, which can balance the intracellular state at low temperatures; and mixtures of organic solvents and water.

[0049] As used herein, the term "therapeutically effective amount" refers to an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dosage level includes the type and severity of the subject, age, sex, drug activity, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, factors including concomitant medications, and other factors well known in the medical field.

[0050] As used herein, the term "treating" a condition or patient means taking steps to obtain a beneficial or desired result, including a clinical result. Beneficial or desired clinical results include, but are not limited to, or eliminating, substantially inhibiting, slowing, or reversing the progression of a disease, condition, or disorder, substantially improving or alleviating the clinical or aesthetic symptoms of a condition, substantially preventing the clinical or aesthetic symptoms of a disease, condition, or disorder, and avoiding harmful or annoying symptoms. Treatment also refers to accomplishing one or more of the following: (a) lessening the severity of the condition; (b) limiting the development of symptoms characteristic of the condition being treated; (c) limiting the worsening of symptoms characteristic of the condition being treated; (d) limiting the recurrence of the condition in patients who previously had the condition; and / or (e) limiting the recurrence of symptoms in patients who were previously free of symptoms of the condition.

[0051] As used herein, the term "prevent" or "preventing" refers to reducing the likelihood of the onset (or recurrence) of a disease, disorder, condition, or associated symptoms. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0053] Figure 1 This is a schematic diagram of the experimental scheme for constructing a rhabdomyolysis-induced acute kidney injury model and then intraperitoneally injecting different volumes of the mixture at different times.

[0054] Figure 2 It is the effect of intraperitoneal injection of different volumes of mixture at different times.

[0055] Figure 3 This is a schematic diagram of the experimental scheme for constructing a rhabdomyolysis-induced acute kidney injury model followed by intraperitoneal injection of different drug combinations.

[0056] Figure 4 It is the effect of different drug combinations injected intraperitoneally.

[0057] Figure 5 It is a schematic diagram of the experimental scheme for administering drug combinations by different administration methods after constructing a rhabdomyolysis-induced acute kidney injury model.

[0058] Figure 6 It is the combined effect of drugs administered in different ways. DETAILED DESCRIPTION

[0059] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0060] As used herein, “containing,” “having,” or “including” include “comprising,” “mainly consisting of,” “substantially consisting of,” and “consisting of;” “mainly consisting of,” “substantially consisting of,” and “consisting of” are subordinate concepts of “containing,” “having,” or “including.”

[0061] The experimental methods used in the following examples are all conventional methods unless otherwise specified, and the reagents, methods and equipment used are all conventional reagents, methods and equipment in the technical field unless otherwise specified.

[0062] Example 1

[0063] Preparation of pharmaceutical preparations:

[0064] Add 800ml of water for injection into the container, add 1.20g of citric acid, 4.30g of sodium citrate, 9g of sodium chloride, and 2.00g of methionine in sequence, stir and dissolve at room temperature for about 30 minutes, add 1mol / L hydrochloric acid or 1mol / L sodium hydroxide to adjust the pH to 6.8-7.2, stir well to completely dissolve, and add water for injection to 1000ml. Filter twice with a 0.22um filter membrane to obtain a mixture for intraperitoneal injection.

[0065] Example 2

[0066] Construction and treatment of animal model of acute kidney injury caused by rhabdomyolysis:

[0067] An induction solution of 50% v / v glycerol / PBS was prepared and injected intramuscularly into the right hind limb of C57BL / 6 mice (male, 10-12 weeks old) at 8 mL / kg to establish a rhabdomyolysis renal disease model (Rh).

[0068] To construct the rhabdomyolysis acute kidney injury model 0h, 3h, such as Figure 1As shown, different total doses, different concentrations of the mixture in Example 1 or 300ul of 0.9% sodium chloride solution (6 mice in each group) were administered by tail vein, oral gavage, and intraperitoneal injection. 12 hours after the acute kidney injury model was established, the mice were sacrificed and peripheral blood was collected to detect creatine kinase, blood creatinine, and urea nitrogen. The kidney tissue was used for H&E pathological staining and renal tubular injury scoring, and polymerase chain reaction (PCR) was used to detect the inflammation expression index IL-6 and the injury index Kim-1.

[0069] The results are as follows Figure 2 As shown (N=6; #<0.05vs con, ##<0.01vs con; *<0.05vs Rh, **<0.01vsRh), according to the results of creatine kinase, blood creatinine, tissue HE staining, PCR Kim-1, and IL-6, it is clear that 0h, 200u1 of the mixture can significantly reduce kidney damage, and increasing the dose cannot further improve kidney damage.

[0070] Example 3

[0071] After establishing the rhabdomyolysis-induced acute kidney injury model, different drug combinations were intraperitoneally injected:

[0072] (1) Prepare a sodium citrate preparation with a concentration matching that of the mixture: add 800 ml of water for injection into a container, add 3.16 g of citric acid, 10.84 g of sodium citrate, and 9 g of sodium chloride in sequence, stir and dissolve at room temperature for about 30 minutes, add 1 mol / L hydrochloric acid or 1 mol / L sodium hydroxide to adjust the pH to 6.8-7.2, stir thoroughly to completely dissolve, and add water for injection to 1000 ml. Filter twice with a 0.22 μm filter membrane to obtain the citric acid preparation.

[0073] (2) Prepare a methionine preparation with a concentration matching that of the mixture: first add 800 ml of water for injection into a container, then add 9 g of sodium chloride and 8.22 g of methionine in sequence, stir and dissolve at room temperature for about 30 minutes, add 1 mol / L hydrochloric acid or 1 mol / L sodium hydroxide to adjust the pH to 6.8-7.2, stir thoroughly to completely dissolve, and add water for injection to 1000 ml. Filter twice with a 0.22 um filter membrane to obtain the methionine preparation.

[0074] like Figure 3 As shown, 0 hours after the model was established in Example 2, 200ul of 0.9% sodium chloride solution, the mixture in Example 1, sodium citrate preparation or methionine preparation was intraperitoneally injected.

[0075] The results are as follows Figure 4As shown (N=6; #<0.05vs con, ##<0.01vs con; *<0.05vs Rh, **<0.01vsRh), according to the results of creatine kinase, blood creatinine, tissue HE staining, PCR Kim-1, and IL-6, it is clear that 200ul of the mixture can significantly reduce renal damage. The improvement effect of using only one of the preparations is limited, and the effect of using the mixture is greater than the effect of adding the two preparations together.

[0076] Example 4

[0077] After establishing the rhabdomyolysis-induced acute kidney injury model, drug combinations were administered by different administration methods:

[0078] like Figure 5 As shown, 200ul of 0.9% sodium chloride solution and the mixture in Example 1 were applied in different ways 0 hours after the model was established in Example 2.

[0079] The results are as follows Figure 6 As shown (N=6; #<0.05vs con, ##<0.01vs con; *<0.05vs Rh, **<0.01vsRh), according to the results of creatine kinase, blood creatinine, tissue HE staining, PCR Kim-1, and IL-6, it is clear that intraperitoneal injection of 200ul of the mixture can significantly reduce renal injury, and other routes can improve renal injury, but the effect is limited. The improvement effect of intraperitoneal injection is better than the sum of the improvements of the other two methods of administration.

[0080] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0081] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: (1) a therapeutically effective amount of sodium citrate and methionine; (2) A pharmaceutically or immunologically acceptable carrier or excipient.

2. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition also includes citric acid and / or sodium chloride.

3. The pharmaceutical composition according to claim 2, characterized in that The mass ratio of the sodium citrate to methionine is 3.08:2.0-4.3:2.

0.

4. The pharmaceutical composition according to claim 3, characterized in that The mass ratio of sodium citrate to methionine is 4.3:2.

0.

5. The pharmaceutical composition according to claim 4, characterized in that The amount of sodium citrate used is 4.3 g; and / or the amount of methionine used is 2.0 g.

6. A pharmaceutical preparation, characterized in that The pharmaceutical preparation comprises the pharmaceutical composition according to any one of claims 1 to 5.

7. The pharmaceutical preparation according to claim 6, characterized in that The administration routes include: intranasal, intrathecal, intraarterial, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, ocular, sublingual, oral, intracerebral and transdermal, or a combination thereof.

8. The pharmaceutical preparation according to claim 7, characterized in that The administration route is intraperitoneal injection.

9. Use of the pharmaceutical composition according to any one of claims 1 to 5 or the pharmaceutical preparation according to any one of claims 6 to 8 in the preparation of a medicament for preventing and / or treating acute kidney injury and / or diseases and / or symptoms associated with acute kidney injury.

10. The use according to claim 9, characterized in that: The acute kidney injury was induced by rhabdomyolysis.

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