WO2022060121A1 - Pharmaceutical composition for preventing or treating sepsis, acute lung injury, or acute respiratory distress syndrome, containing water-solubilized bile acid - Google Patents

Pharmaceutical composition for preventing or treating sepsis, acute lung injury, or acute respiratory distress syndrome, containing water-solubilized bile acid Download PDF

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WO2022060121A1
WO2022060121A1 PCT/KR2021/012692 KR2021012692W WO2022060121A1 WO 2022060121 A1 WO2022060121 A1 WO 2022060121A1 KR 2021012692 W KR2021012692 W KR 2021012692W WO 2022060121 A1 WO2022060121 A1 WO 2022060121A1
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udca
pharmaceutical composition
water
agent
solubilized
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PCT/KR2021/012692
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French (fr)
Korean (ko)
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송영호
고휘진
조봉상
권경애
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주식회사 아미코젠파마
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Priority claimed from KR1020210122927A external-priority patent/KR102624174B1/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
    • A61K31/573—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics

Definitions

  • the present invention relates to a pharmaceutical composition for the prevention or treatment of sepsis, acute lung injury, or acute respiratory distress syndrome, comprising water-solubilized bile acid. More specifically, the present invention relates to a water-soluble ursodeoxycholic acid (UDCA)-based bile acid, or a pharmaceutically acceptable salt thereof, and an anti-inflammatory or anti-allergic agent, sepsis, acute lung injury, Or it relates to a pharmaceutical composition for preventing or treating acute respiratory distress syndrome.
  • UDCA ursodeoxycholic acid
  • Sepsis is a disease that causes rapid and excessive secretion of inflammatory cytokines due to the body's immune response to pathogens invading from the outside, which causes multiple organ failure and death due to a systemic inflammatory response. It can also cause fatal damage to vital organ functions. In particular, lungs damaged by endotoxin become the cause of acute respiratory distress syndrome and become the most important factor influencing the prognosis of sepsis patients.
  • Sepsis causes 30 million cases of sepsis every year worldwide, and the mortality rate within one month is about 30%. It is estimated that at least 1.7 million adults in the United States develop sepsis each year, resulting in 270,000 deaths. Sepsis is the direct cause of 1 in 3 hospital-related deaths.
  • cytokine storm refers to a phenomenon in which the immune system in the human body is overworked and even normal cells are attacked in order to fight bacteria or viruses infiltrating from the outside. In other words, it is a reaction in which secondary infection symptoms occur as the DNA of normal cells is modified due to excessive secretion of cytokines, proteins secreted by immune cells. This cytokine storm was also pointed out as the main cause of high mortality during the Spanish flu and bird flu in the past. On the other hand, since cytokine storm is a symptom of an excessive immune response, it is more likely to occur in young people with high immunity.
  • Cytokine storms often result in short-term death because they cause a massive inflammatory response and multiple organ damage in the body.
  • MERS-infected patients spread in Korea, patients with worsening conditions appeared in younger age groups without underlying diseases.
  • COVID-19 outbreak that spread around the world from 2019 to 2021, he pointed out the cytokine storm as the main cause of death through a thesis investigating the confirmed cases of covid-19.
  • LPS endotoxin
  • ALI acute lung injury
  • Ursodeoxycholic acid improves and improves the stagnant bile flow after LPS treatment only when pre-administered in an LPS-induced cholestasis animal model (Non-Patent Document 2) , LPS-induced sepsis (LPS-induced sepsis) in the animal model to suppress the cytokine storm and especially to increase the survival rate has not yet been studied.
  • LPS-induced sepsis LPS-induced sepsis
  • UDCA is not known at all regarding the prevention or treatment of acute lung injury disease and acute respiratory distress syndrome, and there is no research on it.
  • Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cytokine storm, acute lung injury disease, or acute respiratory distress syndrome caused by infections such as bacteria or viruses.
  • Another object of the present invention is to provide a pharmaceutical composition that can effectively prevent or treat cytokine storm, acute lung injury disease, or acute respiratory distress syndrome caused by infections such as bacteria or respiratory viruses only by oral administration.
  • Another object of the present invention is to provide a pharmaceutical composition that can effectively prevent or treat cytokine storm, acute lung injury disease, or acute respiratory distress syndrome caused by infections such as bacteria or respiratory viruses without inflammation or side effects.
  • a first formulation comprising a water-solubilized Ursodeoxycholic acid (UDCA)-based bile acid or a pharmaceutically acceptable salt thereof; And a therapeutically acceptable anti-inflammatory agent or an anti-allergic agent, a second agent; including, and inhibiting the expression of inflammatory cytokines, sepsis prevention or treatment, a pharmaceutical composition is provided.
  • UDCA Ursodeoxycholic acid
  • a first agent comprising a water-solubilized Ursodeoxycholic acid (UDCA) series bile acid or a pharmaceutically acceptable salt thereof; And a therapeutically acceptable anti-inflammatory agent or an anti-allergic agent, a second agent; including, for the prevention or treatment of acute lung injury disease or acute respiratory distress syndrome, a pharmaceutical composition is provided.
  • UDCA Ursodeoxycholic acid
  • the acute lung injury disease or acute respiratory distress syndrome may be caused by bacterial or respiratory viral infection.
  • the respiratory virus is influenza virus (influenza virus), SARS coronavirus (SARS-CoV), MERS coronavirus (MERS-CoV), COVID-19 virus (SARS-CoV-2), It may be at least one selected from adenovirus, parainfluenza virus (PIV), respiratory syncytial virus (RSV), and rhinovirus.
  • influenza virus influenza virus
  • SARS-CoV SARS coronavirus
  • MERS coronavirus MERS coronavirus
  • COVID-19 virus SARS-CoV-2
  • It may be at least one selected from adenovirus, parainfluenza virus (PIV), respiratory syncytial virus (RSV), and rhinovirus.
  • the acute lung injury disease may be one that causes lung inflammation or lung lesions, and reduces the survival rate.
  • the pharmaceutical composition may be in the form of a formulation for oral administration.
  • the water-solubilized UDCA-based bile acid may be included to be administered in a therapeutically effective amount of 5 to 450 mg/kg per day when orally administered.
  • the pharmaceutical composition may be administered for at least 5 days and at least once a day.
  • the water-solubilized UDCA-based bile acid is one selected from water-solubilized ursodeoxycholic acid (UDCA), tauroursodeoxycholic acid (tUDCA), and glycoursodeoxycholic acid (gUDCA). may include more than one.
  • the first agent may include: a first substance including the UDCA-based bile acid; a second substance which is an invert product of at least one water-soluble starch selected from maltodextrin, dextrin, liquid glucose, corn syrup solid, soluble starch, dextran, and combinations thereof; and water, wherein both the first material and the second material remain in a solution state without being precipitated for all pH values within a selected pH value range.
  • the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:13 at a pH value of 6 to 9.
  • the UDCA-based bile acid is included in an amount of 0.01 to 5 parts by weight based on the total weight of the first formulation, and the water-soluble starch inversion product is used in an amount of 1 to 70 parts by weight based on the total weight of the first formulation. may be included.
  • the sum of the concentrations of UDCA-based bile acids including UDCA, tUDCA, and gUDCA for at least 5 minutes in the lung is equal to the concentration of non-UDCA-based bile acids. can be greater than the sum.
  • the UDCA-based bile acid is UDCA
  • the UDCA concentration within 1 hour in lung tissue may be 0.65 ⁇ 1.03 ⁇ g/g tissue or more.
  • the UDCA-based bile acid is tUDCA
  • the tUDCA concentration within 2 hours in lung tissue may be 0.13 ⁇ 0.17 ⁇ g/g tissue or more.
  • the first formulation may be in the form of a liquid, syrup, or dried formulation.
  • the pharmaceutical composition may be administered by nasal or intravenous injection.
  • the second agent may include a steroid agent or a pharmaceutically acceptable salt thereof, and may increase the survival rate.
  • the second agent may be a glucocorticoid type.
  • the second agent may be included to be administered in a therapeutically effective amount of 0.05 to 10 mg/kg per day.
  • the first agent and the second agent may be administered independently of each other.
  • the present invention prevents sepsis by inhibiting excessive production of inflammatory factors TNF-alpha and IL-6, which are inflammatory factors that cause a cytokine storm in an LPS-induced sepsis mouse animal model, including water-soluble ursodeoxycholic acid (UDCA)-based bile acids Or it can be treated.
  • TNF-alpha and IL-6 are inflammatory factors that cause a cytokine storm in an LPS-induced sepsis mouse animal model, including water-soluble ursodeoxycholic acid (UDCA)-based bile acids Or it can be treated.
  • the present invention includes a water-solubilized bile acid and an anti-inflammatory or anti-allergic agent to significantly increase absorption in lung tissue, suppress cytokine expression, and improve survival rate of sepsis caused by infections such as bacteria and respiratory viruses; It can prevent, improve and treat acute respiratory distress syndrome or lung damage.
  • the water-solubilized ursodeoxycholic acid (UDCA)-based bile acid inhibits the excessive production of TNF-alpha and IL-6, which are inflammatory factors that cause cytokine storm, in an LPS-induced sepsis mouse animal model. It can be usefully used in the treatment of sepsis, acute lung injury and acute respiratory distress syndrome by increasing the survival rate during repeated oral administration.
  • UDCA ursodeoxycholic acid
  • the water-solubilized clean aqueous solution of ursodeoxycholic acid (UDCA) can rapidly deliver an effective therapeutic amount of UDCA into the lung tissue only by oral administration, and in an animal model of LPS-induced sepsis, dexamethasone of a steroid preparation is used in combination with a low dose. Since it was found by the present invention to improve the survival rate and lung disease during repeated administration, it can be usefully used in the treatment of sepsis, acute lung injury and acute respiratory distress syndrome.
  • sepsis acute in which water-soluble UDCA, tUDCA (taurodeoxycholic acid) or gUDCA (glycodeoxycholic acid), which are in vivo metabolites of UDCA, can be rapidly delivered in a therapeutically effective amount to the lung tissue without adverse reactions
  • a composition for preventing or treating lung injury disease and acute respiratory distress syndrome may be provided.
  • the composition according to the present invention can achieve the prevention or treatment effect of sepsis, acute lung injury, and acute respiratory distress disease even by oral administration, nasal administration, or intravenous administration, and in particular, oral administration
  • oral administration nasal administration, or intravenous administration, and in particular, oral administration
  • the UDCA-based bile acid and dexamethasone which are active ingredients in the composition of the present invention, are synthetic drugs, the manufacturing cost is low, and thus, it is possible to effectively prevent or treat sepsis, acute lung injury disease, and acute respiratory distress syndrome at a low cost.
  • Example 1 shows whether a clean aqueous solution is generated according to the pH value of the UDCA solution prepared according to Example 1-3 of the present invention.
  • FIG. 7 is a graph showing the measurement results of the initial inflammation-inducing cytokine TNF-alpha (cytokine TNF-alpha) in serum in an LPS-induced sepsis animal model over time.
  • cytokine IL-6 cytokine IL-6
  • Figure 9 shows the survival rate in the case of single repeated administration of UDCA purification and aqueous solubilized UDCA clean aqueous solution in an LPS-induced sepsis animal model.
  • 10 is a graph showing the survival rate in case of repeated administration alone according to the dose of dexamethasone in an LPS-induced sepsis animal model.
  • 11 shows the survival rate according to repeated administration of water-soluble UDCA clean aqueous solution and low-dose dexamethasone in combination in an LPS-induced sepsis animal model.
  • PK pharmacokinetic
  • UDCA-type bile acids such as UDCA, TUDCA, and GUDCA
  • Pharmacokinetic (PK) data showing the amount of change.
  • UDCA-based bile acids a water-solubilized UDCA clean aqueous solution formulation according to the present invention
  • PK Pharmacokinetic
  • PK 15 is pharmacokinetic (PK) data showing changes with time of bile acids transported to the lung tissue after oral administration of AGP600 containing a water-solubilized UDCA-based bile acid according to the present invention.
  • treatment refers to any action in which the symptoms of sepsis, acute lung injury, or acute respiratory distress disease do not worsen, improve, or change advantageously by administration of the composition of the present invention.
  • the term “comprising as an active ingredient” or “comprising a therapeutically active amount” means a composition for preventing or treating sepsis, acute lung injury disease, or acute respiratory distress syndrome, oral administration, and intravenous injection As such, it means to contain enough to exhibit an effect, for example, a preventive effect, a therapeutic effect, and the like.
  • prevention is to treat the drug before the onset of sepsis, acute lung injury or acute respiratory distress disease, and then take oral administration after the onset of the disease, and then at least reduce the parameters related to the condition to be treated, for example, the severity of symptoms. It means any action you do.
  • the term "clear aqueous solution” or “clean aqueous solution” refers to a solution state substantially free of deposits visually.
  • septicemia or acute lung injury disease includes sepsis, acute lung injury disease, or acute respiratory distress syndrome.
  • YSB201, YSB201-xx or AGP600 is a code name for a water-solubilized UDCA clean aqueous solution formulation.
  • ursodeoxycholic acid (UDCA)-based bile acid refers to ursodeoxycholic acid (UDCA), tauro-ursodeoxycholic acid (tUDCA) and glycoursode. Contains glyco-ursodeoxycholic acid (gUDCA).
  • the present inventors have investigated the effect of inhibiting the excessive production of TNF-alpha and IL-6, which are inflammatory factors that cause cytokine storm, after administration of water-solubilized UDCA-based bile acids in a mouse animal model of endotoxin (Lipopolysaccharide; LPS)-induced sepsis, and in particular, anti-inflammatory
  • LPS endotoxin
  • the present invention was completed by confirming an increase in survival rate upon repeated administration by using a low dose of an agent or an anti-allergic agent in combination.
  • a first formulation comprising a water-solubilized Ursodeoxycholic acid (UDCA)-based bile acid or a pharmaceutically acceptable salt thereof; And a therapeutically acceptable anti-inflammatory agent or an anti-allergic agent, a second agent; including, and inhibiting the expression of inflammatory cytokines, sepsis prevention or treatment, a pharmaceutical composition is provided.
  • UDCA Ursodeoxycholic acid
  • a first agent comprising a water-solubilized Ursodeoxycholic acid (UDCA) series bile acid or a pharmaceutically acceptable salt thereof; And a therapeutically acceptable anti-inflammatory agent or an anti-allergic agent, a second agent; including, for the prevention or treatment of acute lung injury disease or acute respiratory distress syndrome, a pharmaceutical composition is provided.
  • UDCA Ursodeoxycholic acid
  • the pharmaceutical composition of the present application is characterized in that it comprises a ursodeoxycholic acid (UDCA) series bile acid solubilized in the first agent.
  • UDCA ursodeoxycholic acid
  • the UDCA-based bile acid may be a UDCA selected from soluble UDCA, water-soluble UDCA derivatives, UDCA salts, and UDCA conjugated with amines, and may be water-solubilized.
  • water-soluble metal salts and water-soluble O-sulfonated bile acids of UDCA may also be included in the soluble UDCA salts.
  • the water-solubilized UDCA-based bile acid may include at least one selected from water-solubilized ursodeoxycholic acid (UDCA), tauroursodeoxycholic acid (tUDCA), and glycoursodeoxycholic acid (gUDCA).
  • UDCA water-solubilized ursodeoxycholic acid
  • tUDCA tauroursodeoxycholic acid
  • gUDCA glycoursodeoxycholic acid
  • tUDCA glycoursodeoxycholic acid
  • tUDCA glycoursodeoxycholic acid
  • gUDCA glycoursodeoxycholic acid
  • Ursodeoxycholic acid is a hydrophilic bile acid, can be administered orally, and has a low concentration of only about 3% of total bile acids in the human body, but is usually present in human bile, It is the only drug approved.
  • the pharmacological actions of UDCA include antioxidant action, anti-inflammatory action, and anti-apoptotic action. These effects are a very important mechanism for the treatment of lung disease, and are more pronounced when UDCA acts as a single molecule. Therefore, the problem is how to absorb and deliver a substance having such an effect well into the lung tissue of the human body.
  • Crystalline UDCA is classified as a skin redness and is an amphipathic molecule having both hydrophilicity and lipophilicity. Therefore, since it is difficult to act as a single molecule of UDCA, the composition of the present invention enables the UDCA crystals to function as a single molecule by solubilizing them in water at a high concentration.
  • UDCA When administered orally in tablets or capsules, which are conventional crystalline UDCA formulations, about 30 to 60% of the total dose is absorbed along the stomach and intestines by nonionic passive diffusion. Only (20% of the ingested dose) is absorbed by the active transport mechanism into the ileus.
  • UDCA When UDCA is absorbed by hepatocytes, it can be conjugated to tUDCA and gUDCA, and tUDCA and gUDCA bound to UDCA are excreted as bile acids secreted in humans by the liver's first-pass clearance, so the blood concentration of UDCA after oral administration is very high. goes low Therefore, no research results have been published that UDCA can be transported into the lung tissue when orally administered with an existing UDCA formulation. none. In order to solve this problem, in order to treat sepsis, acute lung injury, and acute respiratory distress syndrome, UDCA-based bile acids must be efficiently transported to the lung tissue at a therapeutically effective concentration.
  • composition of the present application because of its chemical characteristics, it is classified as enterohepatic circulation and the hepatic first-pass clearance during oral administration is high. It is possible to solve the fundamental problem of crystalline UDCA-based bile acids that could not be transported.
  • the pharmaceutical composition of the present application is formulated into a clean aqueous solution by dissolving UDCA-based bile acids so that the therapeutically active amount of the UDCA-based bile acids can be transported to the lung tissue within a short period of time by oral administration alone, It can be provided as a composition for preventing or treating acute respiratory distress syndrome.
  • the solubility of UDCA in the pharmaceutical composition of the present application may be about 3,000 times or more (0.15 M vs. 0.05 mM) of the solubility of crystalline UDCA in water, and compared to tUDCA, about 300 times or more (0.15 M) vs. 0.45 mM). Accordingly, the present applicant has completed the present invention regarding a pharmaceutical composition for the treatment of sepsis, acute lung injury, or acute respiratory distress syndrome using water-solubilized UDCA-based bile acids.
  • the pharmaceutical composition of the present application rapidly transports the UDCA-based bile acids to the lung tissue in a therapeutically effective amount, thereby inhibiting the overexpression of the initial inflammatory factors TNF-alpha and IL-6, which are the causative agents of the cytokine storm. there is.
  • the pharmaceutical composition of the present disclosure may down-regulate the expression of a protein at the gene level.
  • the pharmaceutical composition of the present application may increase the survival rate as well as inhibition of cytokine expression by combining the second agent, which is a therapeutically acceptable anti-inflammatory agent.
  • the second agent which is a therapeutically acceptable anti-inflammatory agent.
  • the second agent may include a steroid agent or a pharmaceutically acceptable salt thereof. According to the above configuration, it is possible to reduce the dosage of the steroid preparation, which is the second preparation, so that the side effects caused by the steroid preparation can be minimized.
  • Adrenal corticosteroid steroids can cause infections or worsen infections as side effects when used in high doses for a long time, and adrenocortical dysfunction, diabetes, Cushing's syndrome, peptic ulcer, stomach pain, heartburn, depression, insomnia, osteoporosis, muscle pain, fatty liver , edema, high blood pressure, fluid retention, glaucoma, cataracts, blurred vision, thrombosis, acne, pigmentation, thin and fragile skin, rash, and weight gain.
  • treatment such as sepsis or acute respiratory distress, especially in patients with severe coronavirus infection-19 (covid-19), cannot use high doses for a long period of time, and only low doses are prescribed for a short period of time.
  • medical experts said, "When an inflammatory disease such as a cytokine storm occurs, it is correct to administer corticosteroids to block the activity of urgent immune cells to relieve inflammation. We are giving the opinion that caution is necessary because it can have an adverse effect,” he said.
  • the second agent may be a glucocorticoid-based agent.
  • the steroid preparation herein includes dexamethasone or a pharmaceutically acceptable salt thereof, hydrocortisone or a pharmaceutically acceptable salt thereof, cortisone or a pharmaceutically acceptable salt thereof, prednisone ( prednisone) or a pharmaceutically acceptable salt thereof, prednisolone or a pharmaceutically acceptable salt thereof, methylprednisolone or a pharmaceutically acceptable salt thereof, triamcinolone or a pharmaceutically acceptable salt thereof , may be selected from the group consisting of betamethasone or a pharmaceutically acceptable salt thereof.
  • dexamethasone or a pharmaceutically acceptable salt thereof may be suitable for the steroid preparation.
  • the second agent may be included to be administered in a therapeutically effective amount of 0.1 to 10 mg/kg/day.
  • the second formulation is administered orally, administered as an injection, or injected through a vein for adolescents and adults aged 12 years or older and weighing at least 40 kg. 6 mg once a day, a therapeutically effective amount for up to 10 days. Administration may be suitable for the treatment of sepsis, acute lung injury, or acute respiratory distress syndrome with minimal side effects.
  • the therapeutically effective amount of dexamethasone or a pharmaceutically acceptable salt thereof is dexamethasone in the range of about 0.05 to about 1.5 mg/kg/day, while minimizing side effects, in terms of treatment of sepsis, acute lung injury, or acute respiratory distress syndrome.
  • the therapeutically effective amount of dexamethasone or a pharmaceutically acceptable salt thereof may be more suitably in the range of about 0.05 to about 1.0 mg/kg/day, and in another embodiment, the therapeutically effective amount is 0.05 to about 0.5 mg A range of /kg/day may be more suitable, and in another embodiment a therapeutically effective amount may be more suitable in the range of about 0.05 to about 0.25 mg/kg/day, and in another embodiment a therapeutically effective amount is A range from about 0.05 to about 0.15 mg/kg/day may be even more suitable, and a range from about 0.05 to about 0.1 mg/kg/day may be even more suitable .
  • Dexamethasone is a synthetic component of the glucocorticoid family and has anti-inflammatory, anti-allergic, and anti-endotoxic effects, so it is used for inflammatory and autoimmune diseases (Zhong-hua Wang et al., 2013), but when administered, it suppresses inflammation. By overdosing it, it is necessary to consider various side effects such as failure to suppress bacteria or viruses by lowering the immune response, infection, and gastrointestinal bleeding. In particular, when high-dose steroids are used for a long time, they can cause side effects such as infection or worsening of infections due to weakened immunity.
  • dexamethasone can cause various side effects such as depression, insomnia, osteoporosis, muscle pain, fatty liver, edema, high blood pressure, fluid retention, glaucoma, cataract, blurred vision, thrombosis, acne, pigmentation, thin and fragile skin, rash, and weight gain. Therefore, when administering dexamethasone, the various side effects should be considered, and dexamethasone has not yet been approved as a sepsis-only treatment in any country. However, since the pharmaceutical composition of the present application is administered at a low dose as described above, it is possible to prevent problems due to the side effects in advance.
  • the administration of the pharmaceutical composition of the present disclosure may be performed at the same time as the administration of the first agent, the water-solubilized UDCA-based bile acid, and the second agent, or the two agents may be administered in the same or overlapping time period (eg, , at the same time, on the same day or during the same week).
  • the first agent and the second agent may be administered independently of each other in terms of formulation form and/or administration timing.
  • the first agent may be administered orally
  • the second agent may be administered by intravenous/muscular injection. Even when both the first agent and the second agent are orally administered, they may be administered with different administration timings.
  • the acute lung injury disease or acute respiratory distress syndrome may be caused by bacterial or viral infection.
  • the virus may be a respiratory virus, and the respiratory virus is an influenza virus, SARS-CoV, MERS-CoV, COVID-19.
  • Virus SARS-CoV-2
  • adenovirus adenovirus
  • parainfluenza virus PSV
  • RS virus respiratory syncytial virus
  • RSV rhinovirus
  • the respiratory virus may be a SARS coronavirus, a MERS coronavirus, or a COVID-19 virus.
  • the acute lung injury disease may be one that causes lung inflammation or lung lesions, and reduces the survival rate.
  • the pharmaceutical composition of the present application may have a dosage form suitable for administering a therapeutically effective amount of a UDCA family bile acid or a pharmaceutically acceptable salt thereof.
  • composition of the present application may be in the form of a formulation for oral administration, intranasal administration, or intravenous administration.
  • formulation form for oral administration may be more suitable in that it can maximize the convenience of the patient for the treatment of sepsis, acute lung injury disease, and acute respiratory distress syndrome.
  • the UDCA-based bile acid can be efficiently delivered to the lung tissue in a therapeutically active amount without inflammation or side effects, thereby improving lung disease and suppressing cytokine storm.
  • the pharmaceutical composition of the present application when the pharmaceutical composition of the present application is orally administered to a subject, the sum of the concentrations of UDCA-based bile acids, including UDCA, tUDCA, and gUDCA, for at least 5 minutes in the lung is the same as the concentration of non-UDCA-based bile acids. can be greater than the sum.
  • the UDCA-based bile acid is UDCA
  • the UDCA concentration is 0.65 ⁇ within 1 hour in the lung tissue. It can be made to be 1.03 ⁇ g/g tissue or more.
  • the UDCA-based bile acid is UDCA
  • the UDCA concentration of 7.13 ⁇ 2.29 ⁇ g/g tissue can be achieved within 1 hour in lung tissue.
  • the UDCA of the pharmaceutical composition of the present application starts to be distributed in the lung tissue within a short time of 5 to 10 minutes after oral administration, and can rise to Tmax 0.083 ⁇ 0.0 hr, Cmax 7.13 ⁇ 2.29 ⁇ g/g tissue, and for a certain period of time, that is, It can stay for up to about an hour and then wash-out.
  • the UDCA-based bile acid is tUDCA
  • the tUDCA concentration within 2 hours in lung tissue may be 0.13 ⁇ 0.17 ⁇ g/g tissue or more.
  • the water-solubilized UDCA-based bile acid may be included to be administered in a therapeutically effective amount of 5 to 450 mg/kg per day when orally administered.
  • it may be suitable for the dosage form to be in the form of a formulation for administration at least once a day including 50 mg/kg to 450 mg/kg of UDCA or a pharmaceutically acceptable salt thereof as UDCA, , It may be more suitable to have a formulation for administration at least once a day containing UDCA or a pharmaceutically acceptable salt thereof of 50 to 400 mg/kg/time.
  • the dosage form may be in the form of a formulation for administration at least once a day including 50 to 350 mg/Kg of UDCA or a pharmaceutically acceptable salt thereof as UDCA. In another embodiment, it may be more suitable for the dosage form to be in the form of a formulation for administration at least once a day including 50 to 300 mg/Kg/time of UDCA or a pharmaceutically acceptable salt thereof as UDCA. . In another embodiment, it may be more suitable for the dosage form to be in the form of a formulation for administration at least once a day containing 50 to 200 mg/Kg/time of UDCA or a pharmaceutically acceptable salt thereof as UDCA. .
  • the dosage form may be in the form of a formulation for administration at least once a day containing 50 to 150 mg/Kg/time of UDCA or a pharmaceutically acceptable salt thereof as UDCA.
  • the dosage form may be in the form of a formulation for once-a-day administration containing 450 mg/kg/time of UDCA or a pharmaceutically acceptable salt thereof as UDCA.
  • the oral administration interval of the first agent may be administered at intervals of 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days depending on the symptoms.
  • the pharmaceutical composition of the present application may be administered for at least 5 days and at least once a day. In one embodiment, it may be suitable for the pharmaceutical composition to be administered for 5 to 30 days. In another embodiment, the pharmaceutical composition may be more suitable to be administered for at least 10 days and up to 30 days. In another embodiment, it may be more suitable for the pharmaceutical composition to be administered for at least 10 days and up to 20 days.
  • the pharmaceutical composition of the present application may be administered intravenously.
  • the pharmaceutical composition of the present application can rapidly deliver UDCA bile acid to the lungs through intravenous injection, and does not cause problems such as blockage of blood vessels, inflammation, or redness.
  • the pharmaceutical composition may be an intravenous (IV injection) formulation in the lung tissue, and the UDCA-based bile acid of the composition may be delivered to the lung tissue and does not cause skin redness or inflammation in the lung tissue after injection.
  • the water-solubilized UDCA-based bile acid may be included to be administered in a therapeutically effective amount of 1.0 to 100 mg/kg per day when administered intravenously.
  • the pharmaceutical composition herein may be administered intranasally.
  • UDCA bile acid can be rapidly delivered to the lungs through the nasal mucosa through intranasal administration, and inflammation, redness, etc. do not cause problems.
  • the water-solubilized UDCA-based bile acid may be included to be administered in a therapeutically effective amount of 1.0 to 80 mg/kg per day when administered intranasally.
  • it may be suitable to include solubilized UDCA-based bile acid to be administered in a therapeutically effective amount of 5 to 80 mg/kg per day.
  • the first agent may include a first substance including the UDCA-based bile acid; a second substance which is an invert product of at least one water-soluble starch selected from maltodextrin, dextrin, liquid glucose, corn syrup solid, soluble starch, dextran, and combinations thereof; and water, wherein both the first material and the second material remain in a solution state without being precipitated for all pH values within a selected pH value range.
  • UDCA In the aqueous solubilized UDCA-based bile acid, UDCA is stabilized in water together with maltodextrin, a water-soluble starch inversion product, and as a result, the aqueous solubility of pure UDCA molecules can be increased 3,000 times or more.
  • Water-soluble ursodeoxycholic acid is a nonionic molecule with amphiphilic properties due to its molecular nature, and is dissolved in an aqueous solution in the form of a single molecule. Because it is absorbed, the absorption rate of UDCA is dramatically increased.
  • water-solubilized UDCA containing UDCA dissolved in water at a high concentration of up to 60 g/L as a main ingredient is the most ideal multifunctional drug. Or it can prevent or treat an acute lung injury disease.
  • the first material depends on the pH value range.
  • the minimum weight ratio of the second material to the material may vary. If the minimum weight ratio condition is not satisfied, precipitation of the first material may occur.
  • the water-soluble starch compound is maltodextrin
  • the minimum weight ratio of maltodextrin to UDCA is 1:30, 1:25, 1:20, 1:16, 1:15, 1:13, 1:12, 1:6.
  • the amount of high molecular weight, water-soluble starch invert used in the first formulation may be defined as an amount having a desired concentration in the selected ursodeoxycholic acid and solubility in the pH range described herein.
  • the minimum amount of maltodextrin may be equally applied to tUDCA and gUDCA.
  • the pH value of the pharmaceutical composition is 3 to 10
  • the water-soluble starch inversion product is maltodextrin
  • the minimum weight ratio of maltodextrin to the UDCA-based bile acid may be 1:16 to 1:30. .
  • the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:12 at a pH value of 6.1 to 9.6.
  • the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:13 at a pH value of 6 to 9.
  • the pH value of the pharmaceutical composition is 6.5 to 8
  • the water-soluble starch inversion product is maltodextrin
  • the minimum weight ratio of maltodextrin to UDCA may be 1:13 to 1:30.
  • the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:15 at a pH value of 6 to 9.5.
  • the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:20 at a pH value of 5.5 to 9.4.
  • the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:25 at a pH value of 5.1 to 9.6.
  • the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:30 at a pH value of 2.9 to 9.0.
  • the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:30 at a pH value of 2.9 to 10.2.
  • the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:29.8 at a pH value of 2.2 to 10.8.
  • the UDCA-based bile acid may be included in an amount of 0.01 to 5 parts by weight based on the total weight of the first agent.
  • the prevention or treatment effect of sepsis, acute respiratory distress syndrome, or acute lung injury disease may be insignificant, and when it exceeds 5 parts by weight, clean aqueous solution It can be difficult to manufacture.
  • UDCA or maltodextrin does not become a clean aqueous solution and precipitation occurs due to precipitation, it may be difficult to use intravenous or oral administration in lung tissue.
  • UDCA When precipitation occurs, UDCA is not soluble in water and may exist as crystalline UDCA. If it is used for intranasal administration or intrapulmonary injection, pulmonary inflammation is highly likely to occur due to crystalline UDCA. Preparation as a clean aqueous solution is to remove all crystalline UDCA, which causes skin redness, especially in the preparation of lung tissue injections, intranasal administration preparations, and intravenous injection preparations.
  • UDCA is 0.01 to 5 parts by weight, 0.04 to 5 parts by weight, 0.1 to 5 parts by weight, 1 to 5 parts by weight, 2 to 5 parts by weight, 3 to 5 parts by weight based on the total weight of the first agent. parts, 4 to 5 parts by weight, 0.01 to 3 parts by weight, 0.1 to 3 parts by weight, 1 to 3 parts by weight, 2 to 3 parts by weight, 0.01 to 2.5 parts by weight, 0.1 to 2.5 parts by weight, 1 to 2.5 parts by weight can
  • 0.04 to 2.5 parts by weight of the UDCA content may be suitable, 0.1 to 2.5 parts by weight is suitable, and 1 to 2.5 parts by weight may be more suitable with respect to the total weight of the composition. It may be more suitable to be included in an amount of 0.04 to 2.5 parts by weight.
  • the water-soluble starch inversion product may be suitable to include the water-soluble starch inversion product in an amount of 1 to 70 parts by weight based on the total weight of the first formulation.
  • the water-soluble starch inversion product when included in an amount of less than 1 part by weight, UDCA cannot be dissolved in water in an effective amount, so the prevention or treatment effect of sepsis, acute lung injury disease, or acute respiratory distress syndrome may be insignificant, 70 In the case of an excess of parts by weight, precipitation may occur and UDCA or maltodextrin may precipitate out of the aqueous solution, which may cause redness in the lungs.
  • the maltodextrin may contain 1 to 60 parts by weight, 5 to 60 parts by weight, 10 to 60 parts by weight, 20 to 60 parts by weight, 30 to 60 parts by weight, 40 to 60 parts by weight based on the total weight of the composition.
  • the water-soluble starch inversion product of the present application includes carbohydrates obtained directly from partial or incomplete hydrolysis of starch under various pH conditions.
  • Non-limiting examples may be maltodextrin, dextrin, liquid glucose, corn syrup solids (dry powder of liquid glucose), and combinations thereof.
  • the corn syrup solid may be Maltrin ® M200, and the maltodextrin may be Maltrin ® M700, but is not limited thereto, and may be manufactured under the trade name GPC of Grain Processing Corporation, Muscatine, Iowa, USA.
  • the polymer When the water-soluble starch inversion product of the present application is made of a polymer, the polymer may have at least one reducing end and at least one non-reducing end, and may be straight-chain or branched.
  • the molecular weight may be about 100 mass units or more, or 106 mass units or more.
  • the high molecular weight water-soluble starch conversion product may have a molecular weight of 105 mass units or more.
  • the composition may be provided as an aqueous solution comprising a combination of water-soluble starch invert and/or soluble non-starch polysaccharide.
  • the first formulation of the present application may further include a soluble non-starch polysaccharide.
  • the water-soluble non-starch polysaccharide can be obtained under various pH conditions by various hydrolysis or synthesis mechanisms. Non-limiting examples include dextran, guar gum, pectin, indigestible soluble fiber, and the like.
  • the minimum weight ratio of liquid glucose (e.g., corn syrup) to UDCA required to prevent precipitation of the first formulation herein is about 1:25 (i.e., about 12.5 g per 500 mg of UDCA in 100 ml of water, about 25 g per 1 g of UDCA in 200 ml of water), but is not limited thereto.
  • the minimum amount of dry powder of liquid glucose (corn syrup solids, e.g., Maltrin ® M200) required to prevent precipitation is about 30 g per gram of UDCA in 100 ml of water. and may be about 60 g per 2 g of UDCA in 200 ml of water, but is not limited thereto.
  • the minimum amount of soluble non-starch polysaccharide required to prevent precipitation may be about 50 g of guar gum per 500 mg of ursodeoxycholic acid in 100 ml of water, and ursode in 100 ml of water. 80 g of pectin per 500 mg of oxycholic acid.
  • the minimum required amount of high molecular weight water-soluble starch invert or soluble non-starch polysaccharide can be determined mainly by the absolute amount of UDCA in the solution formulation rather than the concentration.
  • the pharmaceutical composition of the present application may further include dietary fiber when formulated for oral administration.
  • dietary fiber include, but are not limited to, guar gum, pectin, psyllium, oat gum, soybean fiber, oat bran, corn hull, cellulose and wheat bran.
  • the pharmaceutical composition of the present application may further include an emulsifying agent and a suspending agent.
  • emulsifiers include guar gum, pectin, acacia, carrageenan, sodium carboxymethyl cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, methyl cellulose, polyvinyl alcohol, povidone, gum tragacanth, xanthan gum and sorbitan. esters.
  • the pharmaceutical composition of the present application may further include pharmaceutically acceptable additives, and pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, Mannitol, syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry, sodium starch glycolate, lead carnauba, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, Sucrose, dextrose, sorbitol and talc and the like can be used.
  • pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, Mannitol, syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry
  • the pharmaceutically acceptable additive is preferably included in an amount of 0.1 to 90% by weight based on the composition, but is not limited thereto.
  • the pharmaceutical composition of the present application may be administered in various parenteral formulations during actual clinical administration, and when formulated, commonly used fillers, extenders, binders, wetting agents, disintegrants, diluents such as surfactants or excipients are used. can be prepared.
  • Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and injections.
  • the pH of the pharmaceutical composition of the present application may be in a range of 2.9 to 10.2, and may be in a clear aqueous solution state in which the composition is dissolved at the pH value.
  • the first agent may be solubilized in water, and may be in a clean aqueous solution state without precipitation at the above pH. That is, the first formulation may be in a clean aqueous solution state without precipitation of UDCA-based bile acids even after several months (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months).
  • the selected pH range that does not precipitate the compound may be, but is not limited to, about pH 2.9 to about pH 10.2, and suitable, but not limited to, about pH 3 to about pH 9, pH 6 to 8 is more suitable, more preferably pH 6.5 to 8.
  • the pH adjusting material is not limited thereto, but HCl, H 3 PO 4 , H 2 SO 4 , HNO 3 , CH 3 COOH, citric acid, malic acid, tartaric acid, lactic acid, phosphate, edetic acid (eidetic acid) ) and alkalis.
  • the nature and mode of use of such pH adjusting substances are well known in the art.
  • the above pH range is any subset of pH levels obtainable in an aqueous system that, depending on the method of administration, is sufficient for the various formulations to be maintained in solution from the formulation, to be injected into lung tissue or to be absorbed into the blood by oral administration. .
  • the composition can be used as a formulation in which the composition according to the invention is in solution without precipitation at the pH levels prevailing in the oral cavity, stomach and intestines.
  • the UDCA family of bile acids remain dissolved under acidic conditions as free, although generally insoluble under acidic conditions.
  • the pharmaceutical composition of the present application may further include a composition in which solubility is maintained when added to the formulation.
  • the pharmaceutical composition is in the form of intrapulmonary injection, oral administration, intranasal injection or intravenous injection, such as sepsis or acute lung injury disease, prevention of sepsis or acute lung injury disease.
  • a solution may be provided.
  • the first formulation may be in the form of a liquid, syrup, or dried formulation.
  • the syrup may be a regular syrup or a thick syrup.
  • the pharmaceutical composition of the present application may be dried and formulated in powder form.
  • the composition in powder form is easy to store and handle, and there is an advantage in that it is easy to prepare a composition in a desired form.
  • the inventor of the present invention established a method for preparing an aqueous solution in a dissolved form at a high concentration by increasing the water solubility of UDCA having the above characteristics through prior research.
  • a clear and clean UDCA aqueous solution stock solution was prepared by including specific water-soluble starch inversion water and water in a certain ratio for UDCA-based bile acids.
  • a method for preparing a clean aqueous solution of water-solubilized UDCA-based bile acids which is the first formulation of the present application, reference may be made to the method disclosed in Korean Patent Publication No. 10-2018-0092886, the entire patent document is incorporated herein by reference.
  • composition of the present application may be administered in combination with other sepsis, acute lung injury disease, or a therapeutic agent for acute respiratory distress syndrome.
  • the therapeutic agent for sepsis, acute lung injury, or acute respiratory distress syndrome may be for intravenous injection.
  • compositions and methods may be used.
  • many of the compounds may be administered with solubilized ursodeoxycholic acid, other compounds may be included.
  • 6.7 g of sodium hydroxide pellets were dissolved in 400 ml of purified water. Then, 60 g of UDCA was dissolved in the sodium hydroxide solution at room temperature under stirring. Then, 360 g of maltodextrin was added little by little to the clear solution and stirred. Then, a preservative was added in an amount suitable for a pharmaceutical formulation to the clear solution obtained while performing sonication at a high throughput (750W, 20kHz), and the pH was adjusted by dropwise addition of HCl. Purified water was added to adjust the total to 1,000 ml. If necessary, the clear solution was filtered through an appropriate filter device.
  • the prepared ursodeoxycholic acid solution formed a clear aqueous solution without visual precipitation at pH 10.3, 9.2, and 6.7, but formed a precipitate at pH 5.4.
  • Dissolve 6.7 g of sodium hydroxide pellets in 400 ml of purified water made it Then, 60 g of UDCA was dissolved in the sodium hydroxide solution at room temperature under stirring. Then, 360 g of maltodextrin was added little by little to the clear solution and stirred.
  • a preservative was added in an amount suitable for a pharmaceutical formulation to the clear solution obtained while performing sonication at a high throughput (750W, 20kHz), and the pH was adjusted by dropwise addition of HCl. Purified water was added to adjust the total to 1,000 ml. If necessary, the clear solution was filtered through an appropriate filter device. This filtration is important for sterilization or removal of impurities from the raw material, but not for removing particulate matter since the solution is already transparent. As shown in Table 1, the prepared ursodeoxycholic acid solution formed a clear aqueous solution without visual precipitation at pH 10.3, 9.2, and 6.7, but formed a precipitate at pH 5.4.
  • Example 1-1 it was prepared according to the same guidelines as in Example 1-1, except that 720 g of maltodextrin as one high molecular weight water-soluble starch inversion product per 60 g of UDCA was used. As shown in Table 1, the prepared UDCA solution formed a clear aqueous solution without visual precipitation at pH 9.6, 7.3, 6.5, and 6.1, but formed a precipitate at pH 5.5.
  • Example 1-1 it was prepared according to the same guidelines as in Example 1-1, except that 750 g of maltodextrin as one high molecular weight water-soluble starch invert per 50 g of UDCA was used. At this time, 5.7 g of sodium hydroxide pellets were dissolved in 400 ml of purified water and then used. As shown in Table 1, the prepared ursodeoxycholic acid solution formed a clear aqueous solution without visual precipitation at pH 9.5, 8.9, 7.9, 7.1, and 6.0. However, a precipitate was formed at pH 5.5. 1 is a photograph showing whether a clean aqueous solution is generated by putting a UDCA solution at each pH value in a test tube.
  • Example 1-1 it was prepared according to the same guidelines as in Example 1-1, except that 350 g of maltodextrin as one high molecular weight water-soluble starch invert per 17.5 g of UDCA was used. At this time, 2.0 g of sodium hydroxide pellets were dissolved in 400 ml of purified water and then used. As shown in Table 1, the prepared UDCA solution formed a clear aqueous solution without visual precipitation at pH 9.4, 7.1, 6.1, and 5.5. However, a precipitate was formed at pH 5.1. 2 is a photograph showing whether a clean aqueous solution is generated by putting the UDCA solution at each pH value in a test tube.
  • Example 1-1 it was prepared according to the same guidelines as in Example 1-1, except that 350 g of maltodextrin as one high molecular weight water-soluble starch invert per 14 g of UDCA was used. At this time, 1.7 g of sodium hydroxide pellets were dissolved in 400 ml of purified water and then used. As shown in Table 1, the prepared UDCA solution formed a clear aqueous solution without visual precipitation at pH 9.6, 6.1, and 5.1. However, a precipitate was formed at pH 4.0. 3 is a photograph showing whether a clean aqueous solution is generated by putting the UDCA solution at each pH value in a test tube.
  • Example 1-1 it was prepared according to the same guidelines as in Example 1-1, except that 750 g of maltodextrin as one high molecular weight water-soluble starch invert per 25 g of UDCA was used. At this time, 2.8 g of sodium hydroxide pellets were dissolved in 400 ml of purified water and then used. As shown in Table 1, the prepared UDCA solution formed a clear aqueous solution without visual precipitation at pH 9.0, 8.0, 7.0, 6.0, 5.1, 4.1, and 2.9. 4 is a photograph showing whether a clean aqueous solution is generated by putting the UDCA solution at each pH value in a test tube.
  • 0.3 g of sodium hydroxide pellets were dissolved in 500 ml of purified water.
  • 1.0 g of UDCA, 0.5 g of tUDCA, and 0.5 g of gUDCA were dissolved in the sodium hydroxide solution at room temperature under stirring.
  • 60 g of maltodextrin was added little by little to the clear solution and stirred.
  • a preservative was added in an amount suitable for a pharmaceutical formulation to the clear solution obtained while performing sonication at a high throughput (750W, 20kHz), and the pH was adjusted by dropwise addition of HCl.
  • Purified water was added to adjust the total to 1,000 ml.
  • the prepared UDCA solution formed a clear aqueous solution without visual precipitation at pH 10.2, 9.0, 8.1, 7.1, 6.1, 5.1, 4.1, 2.9. 5 is a photograph showing whether a clean aqueous solution is generated by putting the UDCA solution at each pH value in a test tube.
  • the prepared UDCA solution formed a clear aqueous solution without visual precipitation at pH 10.8, 9.9, 8.6, 7.5, 6.5, 5.6, 4.7, 3.5, and 2.2. 6 is a photograph showing whether a clean aqueous solution is generated by putting the tUDCA solution at each pH value in a test tube.
  • Solubilized UDCA clean aqueous solution (referred to as test substance and/or codename AGP600)
  • the water-solubilized ursodeoxycholic acid (UDCA) formulation is a clear, clean aqueous solution containing pristine UDCA and water-soluble starch having a low glucose equivalent.
  • UDCA stock solution was prepared.
  • 750 g of maltodextrin as one high molecular weight water-soluble starch inversion product per 25 g of UDCA was used. After dissolving 2.8 g of sodium hydroxide pellets in 100 ml of purified water, 25 g of UDCA was added thereto and stirred to prepare a transparent UDCA solution (solution 1). In another beaker, 400 ml of purified water was prepared, and 750 g of maltodextrin was added thereto, followed by stirring to obtain a transparent maltodextrin solution (solution 2).
  • a preservative was added in an amount suitable for the pharmaceutical formulation as needed, and the pH was adjusted by dropwise addition using hydrochloric acid (pH 6.0 to 7.5), and then the medicinal water was added and adjusted to a total of 1,000 ml. Thereafter, the transparent solution was sterilized by filtration using a 0.2 ⁇ m filterware filtration device under aseptic conditions or heat sterilized at 80° C. to 100° C. The filtration is important for sterilization, but not to remove particulates as the solution is already clear.
  • the prepared aqueous solubilized UDCA solution formed a clear aqueous solution without visual precipitation at pH 12, 10, 9.0, 8.0, 7.0, 6.0, 5.0, 4.0, 3.0, 2.0.
  • the breeding environment is maintained at 23 ⁇ 3 °C, relative humidity of 55 ⁇ 15 %, ventilation frequency of 10-20 times/hr, lighting time of 12 hours (lights at 8 am - lights out at 8 pm), and illuminance of 150-300 Lux. They were bred in room 1 of the animal breeding area of the Gyeonggi Bio Center.
  • Teklad certified irradiated global 18 % protein rodent diet (2918C, ENVIGO, UK) was supplied from Dooyeol Biotech Co., Ltd. (No. 107 Seongbo Plaza, 91, Baumeo-ro, Seocho-gu, Seoul) and allowed to be freely consumed.
  • the rug was used after autoclaving. Contaminant inspection was performed according to Chemon's SOP. Breeding boxes and breeding densities were housed in a polycarbonate breeding box (W 170 x L 235 x H 125 mm) at 1 animal / breeding box during the period of acclimatization, administration and observation. Breeding boxes, rugs and water bottles were exchanged at least once a week.
  • UDCA water-soluble UDCA reducing the cytokine storm-related lethality by alleviating the rapid excessive inflammatory response and the basis of the mechanism of action were secured. If UDCA can inhibit the concentration of major pro-inflammatory cytokines, UDCA and water-soluble UDCA preparations can be used as prophylactic and therapeutic agents in sepsis, acute respiratory distress disease (ARDS) and novel coronavirus (Covid-19) diseases. .
  • ARDS acute respiratory distress disease
  • COvid-19 novel coronavirus
  • composition of the test group is shown in Table 2 below.
  • the solution prepared except for UDCA in the water-solubilized UDCA clean aqueous solution of Example 2 is called a water-solubilized UDCA-vehicle. That is, 500 ml of purified water was prepared in a beaker, and 750 g of maltodextrin was added thereto, followed by stirring to obtain a transparent maltodextrin solution. A preservative may be added to this solution in an amount suitable for pharmaceutical formulations, and the pH is adjusted to between 3.0 and 9.0 by dropwise addition with hydrochloric acid. Thereafter, purified water was added to adjust the total volume to 1,000 ml.
  • the water-solubilized UDCA-vehicle used in this experiment G4-G6 had a UDCA of 0.0mg/ml, a pH of around 6.0-7.0, and no preservatives.
  • water-solubilized UDCA clean aqueous solution stock solution (25 mg/ml as UDCA) was diluted to suit the G4-G6 group or administered to the test subject mice by adjusting the administration volume.
  • the water-solubilized UDCA (test substance) used in G4-G6 was 25 mg/ml, pH 6.0-7.0, and no preservatives were included.
  • the administration method is shown in Table 3 below.
  • the sepsis induction method is to induce sepsis by administering LPS (Lipopolysaccharide, Sigma Aldrich, Cat No. L3024) once intraperitoneally at a dose of 10 mg/kg using a 1 mL syringe 30 minutes after administration of the test substance (DAY 1). did
  • the evaluation method is as follows.
  • TNF-alpha ELISA kit R&D Systems ® , Cat. No.: MTA00B
  • IL- using serum isolated from blood after abdominal vena cava blood collection for each individual 4 hours after LPS administration 6 ELISA kit R&D Systems ® , Cat. No.: M6000B was used for analysis.
  • the cytokine inhibitory effect of the test substance is shown in FIGS. 7 and 8 .
  • this first test was performed to evaluate the effect on the reduction of inflammatory cytokines according to the administered concentration of the test substance (water-soluble UDCA) in a mouse sepsis model.
  • test substance water-soluble UDCA
  • the test group consists of excipient control group (G1), negative control group (G2), test substance (water soluble UDCA) 50 mg/kg administration group (G3), test substance (water solubilization UDCA) 150 mg/kg administration group (G4), test substance (Water-solubilized UDCA) 450 mg/kg administration group (G5), UDCA (crystalline UDCA formulation in white powder form: Where to buy Daewoong Bio, serial number B9007271) administration group (G6), positive control substance oral administration group (G7) and positive control substance It was set as the intraperitoneal administration group (G8). The number of animals per test group was set to 8 for the survival rate evaluation group (G1-G8). As test items, dead animals, observation of general symptoms, and survival rate were measured. The survival rate effect when the test substance (water-soluble UDCA) was administered alone is shown in Table 4 and FIG. 9 . The number in Table 4 is the number of surviving animals.
  • the survival rate of the test substance was rather lower, showing only 12.5% to 25%.
  • the crystalline UDCA formulation G6 administration group also showed an improvement of 37.5% compared to 25% of the water-soluble UDCA administration group, but only showed the survival level of the negative control group, and was lower than the G1 vehicle control group 50%.
  • the present inventors observed the results of the LPS-induced sepsis model experiment performed directly above in more detail.
  • the combination therapy of the test substance water-soluble UDCA
  • the combination therapy with dexamethasone a basic anti-inflammatory drug used as a positive control
  • composition of the primary test group for determining the dose of the positive control material was the negative control group (G1), the positive control group administered with 5 mg/kg (G2), the group administered with 1.5 mg/kg (G3), the group administered with 0.5 mg/kg (G4), 0.15 A mg/kg administration group (G5) and a 0.05 mg/kg administration group (G6) were set.
  • the combined dose of the positive control material was set at 0.15 mg/kg, and the Whether the survival rate is increased was confirmed in the experiments of Examples 3-5.
  • the test results were statistically analyzed by Kaplen-Meier survival analysis. The results are shown in Table 5 and FIG. 10 .
  • the number in Table 5 is the number of surviving animals.
  • Test group consists of negative control group (G1), 5 mg/kg positive control group (G2), 0.15 mg/kg positive control group (G3), 0.15 mg/kg excipient and positive control substance combination group (G4), test Substance (water-solubilized UDCA) 100 mg/kg and positive control substance 0.15 mg/kg combined administration group (G5), test substance (water-solubilized UDCA) 200 mg/kg and positive control substance 0.15 mg/kg combination administration group (G6), test Substance (water-soluble UDCA) 300 mg/kg and positive control substance 0.15 mg/kg combined administration group (G7) and test substance 0.5 mg/head nasal administration group (G8) were set. The number of animals per test group was all set to 8.
  • test substance water-soluble UDCA
  • water-soluble UDCA 200 mg/kg and positive control substance 0.15 mg/kg group
  • 2 cases died on DAY 3 and 1 case on DAY 5.
  • test substance water-soluble UDCA
  • 300 mg/kg and positive control substance 0.15 mg/kg group G7
  • 1 case died on DAY 2, 1 case on DAY 3, and 1 case on DAY 4.
  • Two cases died on DAY 3 in the test substance water-soluble UDCA) 0.5 mg/head/time nasal administration group (G8).
  • a survival rate of 75.00% was observed in the UDCA) 0.5 mg/head/time nasal administration group (G8).
  • the survival rate (G3) was 37.50%, whereas when administered in combination with a test substance, when water-solubilized UDCA was administered at 100 mg/kg (G5), the survival rate was 75.00%, surprisingly, the survival rate was 200 % (2.14 times) increased, and when the water-solubilized UDCA was administered at 200 mg/kg (G6), it was also increased by 166% (1.67 times) to 62.5%.
  • solubilized UDCA is very valuable for increasing the survival rate in the treatment of sepsis disease, ARDS, and coronavirus disease-19 (Covid-19) disease when co-administered with dexamethasone can be used
  • dexamethasone can only be treated for a short period of time in the treatment of severe coronavirus infection-19 (Covid-19) disease.
  • the results of the secondary test are shown in Table 6 and graphs of FIG. 11 .
  • the test results were statistically analyzed by Kaplen-Meier survival analysis, and the number in Table 6 is the number of surviving animals.
  • test substance water-soluble UDCA
  • water-soluble UDCA water-soluble UDCA
  • test substance water-soluble UDCA
  • water-soluble UDCA suppressed the expression levels of TNF-alpha and IL-6, the causative agents of cytokine storm, and the survival rate when co-administered with dexamethasone, a low-dose steroid, was approximately 200 % (2 times) increased.
  • water-soluble UDCA significantly inhibits the expression of cytokines such as TNF-alpha and IL-6 and increases the survival rate when combined with steroids such as mexamethasone. . In particular, it can increase the survival rate in the treatment of acute respiratory distress caused by acute lung injury in COVID-19.
  • lung tissue samples were collected for each hour, and drug components in living tissues were extracted using an organic solvent, and the concentration was quantitatively analyzed using an HPLC fluorescence detector. Sample collection was performed 0, 5, 10, 30 minutes and 1, 2, 4, 10, 24, 48, 72 hours after oral administration, and biological samples were collected from 4 mice at each hour. In the case of the control group that had fasted and fed the same as the test group, biological samples of lung tissue were collected after 0, 4, 10, 24, 48, and 72 hours.
  • AGP600 undiluted aqueous solution of UDCA in Example 2; UDCA concentration: 25 mg/ml was used as a test substance.
  • gUDCA glycoursodeoxycholic acid
  • tUDCA tauroursodeoxycholic acid
  • UDCA ursodeoxycholic acid
  • GCA Glycocholic acid hydrate
  • TCA Taurocholic acid sodium salt hydrate
  • CA Cholic acid
  • GCDCA Glycochenodeoxycholic acid
  • TCDCA Taurochenodeoxycholic acid
  • GDCA Glycodeoxycholic acid
  • TDCA Taurodeoxycholic acid
  • CDCA Chenodeoxycholic acid
  • DCA Deoxycholic acid
  • GLCA Glycolithocholic acid sodium salt
  • TLCA Glycolithocholic acid sodium salt
  • TLCA Glycolithocholic acid sodium salt
  • TLCA Glycolithocholic acid sodium salt
  • LCA Lithocholic acid
  • the analysis equipment is 2695 Alliance HPLC (high performance liquid chromatography) equipment of Water Corporation of the United States, and BilePak II column (4.6125 mm, JASCO, Japan) and EnzymePak 3 ⁇ -HSD column (4.635 mm, JASCO, Japan) of JASCO of Japan are used. used together.
  • the dose for each individual was converted based on the measured body weight, and a sonde was attached to a disposable syringe for oral administration.
  • mice C57BL/6 females
  • mice having a body weight range of 16 to 18 g were received and acclimatized for 7 days before being used in the experiment.
  • animals within ⁇ 20% of the total average body weight were used.
  • the place of purchase is Coretech Co., Ltd. (Pyeongtaek, Gyeonggi-do, Korea).
  • the environment of the animal room was set at 19 ⁇ 25°C, 40 ⁇ 60% humidity, and 150-300Lux illuminance.
  • composition and administration plan of the control and test groups are shown in Table 7.
  • test results were summarized using Microsoft Excel 2010, and additional pharmacokinetic analysis was performed on the results of concentration analysis in lung tissue using the Pharsight WinNonlin 7.0 program (Certara, USA).
  • Pharsight WinNonlin 7.0 program (Certara, USA).
  • p ⁇ 0.05 value was regarded as a statistically significant value and statistically processed, and a graph was prepared.
  • water-solubilized UDCA was rapidly delivered to the lungs, and a constant concentration was maintained in the lungs for 1 hour. It was found that the water-solubilized UDCA aqueous solution formulation of the present invention can transport UDCA in a therapeutically active amount to the lungs only by oral administration, and can maintain the delivered UDCA for 1 hour or more without immediately disappearing. Subsequently, tUDCA, a metabolite of UDCA, increased in the lung tissue at a time when UDCA gradually disappeared after Tmax and was maintained for a certain period of time, as shown in FIG. 13 .
  • UDCA and tUDCA which are UDCA-family bile acids, are known to have a cytoprotection function.
  • water-soluble UDCA of the present invention was orally administered, it was confirmed that the UDCA-based bile acids were present in the lung tissue at a higher concentration than other non-UDCA-based bile acids including cytotoxic hydrophobic bile acids for 1 hour or more ( 15).
  • UDCA in lung tissue of test group mice was the highest concentration (Cmax) in lung tissue at Tmax 0.083 hours. It showed 7.13 ⁇ 2.29 ⁇ g/g tissue, and AUC (hr ⁇ g/g tissue) was 12.79 ⁇ 8.49.
  • the AUC ratio which is the ratio of the sum of AUCs of UDCA-family bile acids (UDCA, tUDCA, and gUDCA) divided by the sum of AUCs of other bile acids, was 1.10.
  • tUDCA an in vivo metabolite of UDCA and a cytoprotective effect
  • tUDCA in lung tissue showed the highest concentration (Cmax) in lung tissue (Cmax) of 4.36 ⁇ 0.85 ⁇ g/g tissue at Tmax 1.13 ⁇ 0.31 hours, and AUC (hr ⁇ g/g tissue) was 28.31 ⁇ 10.57.
  • the average UDCA content within 1 hour after oral administration in lung tissue was 0.65 ⁇ 1.03 ⁇ g/g tissue or more, and the average TUDCA content within 2 hours after oral administration was 0.13 ⁇ 0.17 ⁇ g/g tissue.
  • Non-Patent Document 1 Mosamichi Ota, et al., 1977
  • UDCA containing 14 C radioactive isotope was administered to rats at 30 mg/Kg at 30 mg/Kg once a day at intervals of 7
  • the distribution in tissues was examined, and no radioisotope was detected in the lungs. That is, it was confirmed that UDCA that is not solubilized hardly transports UDCA into the lung tissue when taken orally.
  • UDCA is first delivered to the lung tissue at a high concentration, and then, when dexamethasone, a type of steroid, is administered in combination, the function of damaged lung cells is restored more effectively than when dexamethasone is administered alone, resulting in survival rate. and lung damage can be improved.
  • the water-soluble UDCA-based bile acid directly protects the damaged lung cells in the lung tissue, and is caused by bacteria or respiratory viruses such as influenza virus, SARS-CoV, and MER.
  • MERS-CoV COVID-19 virus (SARS-CoV-2), adenovirus, parainfluenza virus (PIV), respiratory syncytial virus (RSV), rhinovirus ( It can be seen that it can have a direct protective effect on sepsis, acute respiratory distress syndrome, or lung injury caused by infection with rhinovirus).
  • UDCA In order to transport UDCA to the lung tissue at a high concentration, UDCA was solubilized. Surprisingly, the present inventors found that UDCA could be transported to the lungs in a therapeutically active amount only by oral administration. In addition, UDCA does not disappear from the lungs immediately and stays for about 1 hour to exert its efficacy, and tUDCA, a metabolite of UDCA, is sequentially transported to the lungs and stayed for a total of 2 hours. It has been found that the prevention and treatment of injuries are effective.
  • UDCA is delivered to the lungs at a high concentration and inhibits the early proinflammatory factors TNF-a and IL-6 known to cause a cytokine storm, while at the same time effectively restoring the function of damaged lung cells, thereby improving survival rate and lung disease. show that it can be improved.
  • ursodeoxycholic acid a type of bile acid mainly used for liver treatment, suppresses cytokine storms caused by sepsis, acute respiratory distress or acute lung injury caused by infections such as bacteria or respiratory viruses, and , it was confirmed through LPS-induced sepsis animal model experiment whether it can improve the survival rate and lung disease.
  • oral administration of UDCA had an inhibitory effect on the overexpression of TNF-alpha and IL-6, which are the causative agents of the cytokine storm.
  • dexamethasone a synthetic corticosteroid (corticosteroid) preparation having an anti-inflammatory action.
  • dexamethasone showed a 100% survival rate only when high-dose repeated administration, as reported in previous papers.
  • low-dose repeated administration When not administered, the survival rate was only increased by 36.5% compared to the 0% survival rate.
  • the present inventors have devised a combination of repeated oral administration of a low dose of UDCA and dexamethasone in an animal model of LPS-induced sepsis for the development of a therapeutic agent for sepsis, acute lung injury, and acute respiratory distress.
  • the present inventors combined UDCA and a low dose of dexamethasone, a steroid, in combination with a repeated administration method to administer LPS-induced sepsis in an animal model.
  • the survival rate of UDCA alone increased from 25.0 to 37.5% to 75%
  • the survival rate could be increased by more than 200% (2 times) more than 37.5% when administered with a low dose of dexamethasone alone, 75% when administered in combination with water-soluble UDCA. It was also found that, at the same time, weight was restored and lung inflammation was greatly improved.

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Abstract

The present invention relates to a pharmaceutical composition for preventing or treating sepsis, acute lung injury, or acute respiratory distress syndrome, containing water-solubilized bile acid. More specifically, the present invention relates to a pharmaceutical composition for preventing or treating sepsis, acute lung injury, or acute respiratory distress syndrome, involving co-administration of water-solubilized ursodeoxycholic acid (UDCA)-based bile acid or a pharmaceutically acceptable salt thereof and an anti-inflammatory or anti-allergic agent.

Description

수가용화된 담즙산을 포함하는 패혈증, 급성 폐손상 질환, 또는 급성 호흡곤란증후군의 예방 또는 치료용 약학 조성물Pharmaceutical composition for prevention or treatment of sepsis, acute lung injury disease, or acute respiratory distress syndrome comprising water-solubilized bile acid
본 발명은 수가용화된 담즙산을 포함하는 패혈증, 급성 폐손상 질환, 또는 급성 호흡곤란증후군의 예방 또는 치료용 약학 조성물에 관한 것이다. 더욱 상세하게, 본 발명은 수가용화된 우르소데옥시콜산 (ursodeoxycholic acid, UDCA) 계열 담즙산, 또는 이의 약학적으로 허용가능한 염과 항염증 또는 항알러지 제제를 병용 투여하는, 패혈증, 급성 폐손상 질환, 또는 급성 호흡곤란증후군의 예방 또는 이로 인한 치료용 약학 조성물에 관한 것이다. The present invention relates to a pharmaceutical composition for the prevention or treatment of sepsis, acute lung injury, or acute respiratory distress syndrome, comprising water-solubilized bile acid. More specifically, the present invention relates to a water-soluble ursodeoxycholic acid (UDCA)-based bile acid, or a pharmaceutically acceptable salt thereof, and an anti-inflammatory or anti-allergic agent, sepsis, acute lung injury, Or it relates to a pharmaceutical composition for preventing or treating acute respiratory distress syndrome.
패혈증은 외부에서 침입한 병원균에 대한 인체의 면역반응에 의해 염증성 사이토카인의 급격한 과다한 분비가 일어나고, 이로 인해 전신성 염증 반응으로 다장기부전, 사망을 초래하는 질병으로 치사율이 매우 높으며 종종 폐나 신장과 같은 주요 장기 기능에 치명적인 손상을 초래하기도 한다. 특히 내독소로 인하여 손상받은 폐(Lung)는 급성 호흡곤란증후군의 원인이 되어 패혈증 환자의 예후를 좌우하는 가장 중요한 인자가 된다.Sepsis is a disease that causes rapid and excessive secretion of inflammatory cytokines due to the body's immune response to pathogens invading from the outside, which causes multiple organ failure and death due to a systemic inflammatory response. It can also cause fatal damage to vital organ functions. In particular, lungs damaged by endotoxin become the cause of acute respiratory distress syndrome and become the most important factor influencing the prognosis of sepsis patients.
패혈증으로 인해 매년 세계적으로 연간 3,000만명의 환자가 발생하고 한달내 사망률이 30%에 달할 정도이다. 미국에선 해마다 최소 170만 명의 성인이 패혈증을 일으켜 27만 명이 사망하는 것으로 알려졌다. 병원에서 사망하는 환자 3명 중 1명꼴은 패혈증이 직접 원인이라고 한다.Sepsis causes 30 million cases of sepsis every year worldwide, and the mortality rate within one month is about 30%. It is estimated that at least 1.7 million adults in the United States develop sepsis each year, resulting in 270,000 deaths. Sepsis is the direct cause of 1 in 3 hospital-related deaths.
패혈증의 주된 원인은 조절 불가능한 염증반응이다. "사이토카인 폭풍(cytokine storm)" 이라고도 불리우며, 외부에서 침투한 세균이나 바이러스에 대항하기 위해 인체 내에서 면역작용이 과다하게 이뤄지면서 정상 세포까지 공격하는 현상을 말한다. 즉, 면역세포가 분비하는 단백질인 사이토카인의 과다 분비로 정상 세포들의 DNA가 변형되면서 2차 감염 증상이 일어나는 반응이다. 이 사이토카인 폭풍은 과거 스페인 독감·조류독감 등이 유행할 때 높은 사망률의 주된 원인으로 지목되기도 했다. 한편, 사이토카인 폭풍은 면역반응의 과잉으로 나타나는 증상이기 때문에 면역력이 높은 젊은 층에서 발생할 확률이 더 높다. 사이토카인 폭풍은 신체에 대규모 염증반응과 다발성 장기손상을 일으키기 때문에 단기간 사망에 이르는 경우가 많다. 2015년 국내에서 메르스(MERS) 감염자가 확산될 때 기저질환이 없는 젊은 연령대에서 상태 악화를 보이는 환자들이 나타나자, 의료계에서는 이 현상의 원인으로 사이토카인 폭풍을 거론하기도 했다. 또 2019~2021년 전 세계로 확산된 코로나19 사태에서도 covid-19 확진자를 조사한 논문을 통해 사이토카인 폭풍을 주요 사인(死因)으로 지목한 바 있다.The main cause of sepsis is an uncontrolled inflammatory response. Also called "cytokine storm", it refers to a phenomenon in which the immune system in the human body is overworked and even normal cells are attacked in order to fight bacteria or viruses infiltrating from the outside. In other words, it is a reaction in which secondary infection symptoms occur as the DNA of normal cells is modified due to excessive secretion of cytokines, proteins secreted by immune cells. This cytokine storm was also pointed out as the main cause of high mortality during the Spanish flu and bird flu in the past. On the other hand, since cytokine storm is a symptom of an excessive immune response, it is more likely to occur in young people with high immunity. Cytokine storms often result in short-term death because they cause a massive inflammatory response and multiple organ damage in the body. In 2015, when MERS-infected patients spread in Korea, patients with worsening conditions appeared in younger age groups without underlying diseases. In addition, in the COVID-19 outbreak that spread around the world from 2019 to 2021, he pointed out the cytokine storm as the main cause of death through a thesis investigating the confirmed cases of covid-19.
1980년대 초반 이후로 수많은 패혈증 치료 후보 약물들에 대한 무작위 배정 비교 임상 시험이 수행되어 왔으나 아직까지도 패혈증 치료제로 허가받은 약물이 없는 실정이다. 즉 패혈증으로 인한 빠르고 과도한 염증을 조절할 수 있는 강력한 표적치료제가 없어 치료에 어려움을 겪고 있다. 현재 세계 어느 나라에서도 보건당국으로부터 의약품으로 승인받은 패혈증 전용 치료제는 아직 개발되지 않았다.Since the early 1980s, randomized comparative clinical trials have been conducted on numerous drug candidates for the treatment of sepsis, but there is still no drug approved for the treatment of sepsis. In other words, there is no strong targeted therapy that can control the rapid and excessive inflammation caused by sepsis, so it is difficult to treat. Currently, in any country in the world, a treatment for sepsis that has been approved as a medicine by the health authorities has not yet been developed.
내독소 (Lipopolysaccharide; LPS) 유발 패혈증 마우스 실험 동물모델은 복강 내로 내독소(LPS)를 주입하여 빠른 과도한 염증 반응으로 면역 세포들이 초기 염증유발 사이토카인인 TNF-alpha, IL-6 등을 과다 생성하여 사이토카인 폭풍(cytokine storm)을 일으키는 대표적인 패혈증 동물모델이다. LPS로 유발된 동물모델은 특히 급성 폐손상(Acute Lung Injury; ALI) 마우스 모델로도 잘 알려져 있으며, ALI 병리 연구 및 신약 개발에 널리 사용되고 있다(비특허문헌 1).In a mouse experimental animal model of endotoxin (LPS)-induced sepsis, endotoxin (LPS) was injected intraperitoneally to cause rapid excessive inflammatory response and immune cells over-produced early inflammatory cytokines such as TNF-alpha and IL-6. It is a representative animal model of sepsis that causes a cytokine storm. LPS-induced animal models are particularly well known as acute lung injury (ALI) mouse models, and are widely used in ALI pathology research and drug development (Non-Patent Document 1).
우르소데옥시콜산(UDCA)이 LPS-유발 담즙정체(cholestasis) 동물모델에서 미리 투여한 경우에만 LPS 처리 후 정체된 담즙산의 흐름(bile flow)을 향상시키고 개선한다는 발표는 있지만(비특허문헌 2), LPS-유도 패혈증(LPS-induced sepsis) 동물모델에서 사이토카인 폭풍을 억제하고 특히 생존율을 증가시킨다는 연구결과는 아직 없다. 특히 UDCA가 급성 폐손상 질환 및 급성 호흡곤란증후군에 대한 예방 또는 치료와 관련해서는 전혀 알려진 바 없으며, 이에 대한 연구도 전무한 상황이다.Ursodeoxycholic acid (UDCA) improves and improves the stagnant bile flow after LPS treatment only when pre-administered in an LPS-induced cholestasis animal model (Non-Patent Document 2) , LPS-induced sepsis (LPS-induced sepsis) in the animal model to suppress the cytokine storm and especially to increase the survival rate has not yet been studied. In particular, UDCA is not known at all regarding the prevention or treatment of acute lung injury disease and acute respiratory distress syndrome, and there is no research on it.
[선행기술문헌][Prior art literature]
[특허문헌][Patent Literature]
1: 한국공개특허 제10-2018-0092886호(우르소데옥시콜산을 함유하는 시각장애 예방 또는 치료용 조성물, 2018년 08월 20일 공개)1: Korea Patent Publication No. 10-2018-0092886 (composition for preventing or treating visual impairment containing ursodeoxycholic acid, published on August 20, 2018)
2: 한국공개특허 제10-2018-0036580호(수가용화(水加溶化)된 우르소데옥시콜산을 함유하는 염증성 피부질환 또는 중증 소양증 예방 또는 치료용 조성물, 2018년 04월 09일 공개)2: Korean Patent Laid-Open Patent No. 10-2018-0036580 (Composition for preventing or treating inflammatory skin disease or severe pruritus containing water-solubilized ursodeoxycholic acid, published on April 09, 2018)
[비특허문헌][Non-patent literature]
1: Ji-Sun Hwang et al., Glucosamine improves survival in a mouse model of sepsis and attenuates sepsis-induced lung injury and inflammation. J. Biol. Chem. 294(2) (2019). 608-6221: Ji-Sun Hwang et al., Glucosamine improves survival in a mouse model of sepsis and attenuates sepsis-induced lung injury and inflammation. J. Biol. Chem. 294(2) (2019). 608-622
2: Randa H. Ainosah et al., The effects of ursodeoxycholic acid on sepsis-induced cholestasis management in an animal model. Journal of Taibah University Medical Sciences (2020) 15(4), 312e3202: Randa H. Ainosah et al., The effects of ursodeoxycholic acid on sepsis-induced cholestasis management in an animal model. Journal of Taibah University Medical Sciences (2020) 15(4), 312e320
본 발명의 목적은 패혈증 예방 또는 치료용 약학 조성물을 제공하는 데 있다.It is an object of the present invention to provide a pharmaceutical composition for preventing or treating sepsis.
본 발명의 다른 목적은 세균 또는 바이러스 등의 감염에 의한 사이토카인 폭풍, 급성 폐손상 질환, 또는 급성 호흡곤란증후군 예방 또는 치료용 약학 조성물을 제공하는 데 있다.Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cytokine storm, acute lung injury disease, or acute respiratory distress syndrome caused by infections such as bacteria or viruses.
본 발명의 또 다른 목적은 경구투여만으로 세균 또는 호흡기 바이러스 등의 감염에 의한 사이토카인 폭풍, 급성 폐손상 질환, 또는 급성 호흡곤란증후군을 효과적으로 예방 또는 치료할 수 있는 약학 조성물을 제공하는 데 있다.Another object of the present invention is to provide a pharmaceutical composition that can effectively prevent or treat cytokine storm, acute lung injury disease, or acute respiratory distress syndrome caused by infections such as bacteria or respiratory viruses only by oral administration.
본 발명의 또 다른 목적은 염증이나 부작용 없이 세균 또는 호흡기 바이러스 등의 감염에 의한 사이토카인 폭풍, 급성 폐손상 질환, 또는 급성 호흡곤란증후군을 효과적으로 예방 또는 치료할 수 있는 약학 조성물을 제공하는 데 있다.Another object of the present invention is to provide a pharmaceutical composition that can effectively prevent or treat cytokine storm, acute lung injury disease, or acute respiratory distress syndrome caused by infections such as bacteria or respiratory viruses without inflammation or side effects.
본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명, 청구범위 및 도면에 의해 더욱 명확하게 된다.Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.
본원의 일 측면에 의하면, 수가용화된 우르소데옥시콜산(Ursodeoxycholic acid, UDCA) 계열 담즙산 또는 이의 약학적으로 허용가능한 염을 포함하는 제1 제제; 및 치료학적으로 허용가능한 항염증 제제 또는 항알러지 제제인 제2 제제;를 포함하고, 염증성 사이토카인 발현을 억제하는, 패혈증 예방 또는 치료용, 약학 조성물이 제공된다.According to one aspect of the present application, a first formulation comprising a water-solubilized Ursodeoxycholic acid (UDCA)-based bile acid or a pharmaceutically acceptable salt thereof; And a therapeutically acceptable anti-inflammatory agent or an anti-allergic agent, a second agent; including, and inhibiting the expression of inflammatory cytokines, sepsis prevention or treatment, a pharmaceutical composition is provided.
본원의 다른 측면에 의하면, 수가용화된 우르소데옥시콜산(Ursodeoxycholic acid, UDCA) 계열 담즙산 또는 이의 약학적으로 허용가능한 염을 포함하는 제1 제제; 및 치료학적으로 허용가능한 항염증 제제 또는 항알러지 제제인 제2 제제;를 포함하고, 급성 폐손상 질환 또는 급성 호흡곤란증후군 예방 또는 치료용, 약학 조성물이 제공된다.According to another aspect of the present application, a first agent comprising a water-solubilized Ursodeoxycholic acid (UDCA) series bile acid or a pharmaceutically acceptable salt thereof; And a therapeutically acceptable anti-inflammatory agent or an anti-allergic agent, a second agent; including, for the prevention or treatment of acute lung injury disease or acute respiratory distress syndrome, a pharmaceutical composition is provided.
본원의 일 실시예에 의하면, 급성 폐손상 질환 또는 급성 호흡곤란증후군은 세균 또는 호흡기 바이러스 감염에 의한 것일 수 있다.According to an embodiment of the present application, the acute lung injury disease or acute respiratory distress syndrome may be caused by bacterial or respiratory viral infection.
본원의 일 실시예에 의하면, 상기 호흡기 바이러스는 인플루엔자 바이러스(influenza virus), 사스 코로나바이러스(SARS-CoV), 메르스 코로나바이러스(MERS-CoV), COVID-19 바이러스(SARS-CoV-2), 아데노바이러스(adenovirus), 파라인플루엔자 바이러스 (parainfluenza virus, PIV), RS 바이러스(respiratory syncytial virus, RSV), 및 라이노바이러스(rhinovirus) 중에서 선택된 1종 이상일 수 있다.According to an embodiment of the present application, the respiratory virus is influenza virus (influenza virus), SARS coronavirus (SARS-CoV), MERS coronavirus (MERS-CoV), COVID-19 virus (SARS-CoV-2), It may be at least one selected from adenovirus, parainfluenza virus (PIV), respiratory syncytial virus (RSV), and rhinovirus.
본원의 일 실시예에 의하면, 상기 급성 폐손상 질환은 폐 염증 또는 폐 병변을 일으키고, 생존율을 감소시키는 것일 수 있다.According to an embodiment of the present application, the acute lung injury disease may be one that causes lung inflammation or lung lesions, and reduces the survival rate.
본원의 일 실시예에 의하면, 약학 조성물은 경구투여용 제제 형태일 수 있다.According to an embodiment of the present application, the pharmaceutical composition may be in the form of a formulation for oral administration.
본원의 일 실시예에 의하면, 상기 수가용화된 UDCA 계열 담즙산은 경구투여 시 1일 5 내지 450mg/kg의 치료학적 유효량으로 투여되도록 포함될 수 있다.According to an embodiment of the present application, the water-solubilized UDCA-based bile acid may be included to be administered in a therapeutically effective amount of 5 to 450 mg/kg per day when orally administered.
본원의 일 실시예에 의하면, 상기 약학 조성물은 5일 이상 그리고 1일 1회 이상 투여되는 것일 수 있다.According to an embodiment of the present application, the pharmaceutical composition may be administered for at least 5 days and at least once a day.
본원의 일 실시예에 의하면, 상기 수가용화된 UDCA 계열 담즙산은 수가용화된 우르소데옥시콜산(UDCA), 타우로우르소데옥시콜산(tUDCA) 및 글리코우르소데옥시콜산(gUDCA) 중에서 선택되는 1종 이상을 포함할 수 있다.According to an embodiment of the present application, the water-solubilized UDCA-based bile acid is one selected from water-solubilized ursodeoxycholic acid (UDCA), tauroursodeoxycholic acid (tUDCA), and glycoursodeoxycholic acid (gUDCA). may include more than one.
본원의 일 실시예에 의하면, 상기 제1 제제는, 상기 UDCA 계열의 담즙산을 포함하는 제1 물질; 말토덱스트린, 덱스트린, 액상 포도당, 옥수수 시럽 고체, 가용성 전분, 덱스트란 및 이들의 조합에서 선택되는 1종 이상의 수가용성 전분 전화물인 제2 물질; 및 물;을 포함하고, 상기 제1 물질 및 상기 제2 물질이 모두, 선택된 pH 값 범위 내의 모든 pH 값에 대해 침전되지 않고 용액상태로 남아 있는 것일 수 있다.According to an embodiment of the present application, the first agent may include: a first substance including the UDCA-based bile acid; a second substance which is an invert product of at least one water-soluble starch selected from maltodextrin, dextrin, liquid glucose, corn syrup solid, soluble starch, dextran, and combinations thereof; and water, wherein both the first material and the second material remain in a solution state without being precipitated for all pH values within a selected pH value range.
본원의 일 실시예에 의하면, 상기 수가용성 전분 전화물은 말토덱스트린이고, 상기 UDCA 계열의 담즙산에 대한 상기 말토덱스트린의 최소 중량비는 pH 값 6 내지 9에서 1:13일 수 있다.According to an embodiment of the present application, the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:13 at a pH value of 6 to 9.
본원의 일 실시예에 의하면, 상기 UDCA 계열의 담즙산은 제1 제제의 총 중량에 대하여 0.01 ~ 5 중량부로 포함되고, 상기 수가용화 전분 전화물은 제1 제제의 총 중량에 대하여 1 ~ 70 중량부로 포함될 수 있다.According to an embodiment of the present application, the UDCA-based bile acid is included in an amount of 0.01 to 5 parts by weight based on the total weight of the first formulation, and the water-soluble starch inversion product is used in an amount of 1 to 70 parts by weight based on the total weight of the first formulation. may be included.
본원의 일 실시예에 의하면, 약학 조성물은 대상에 경구투여 시 폐(lung)에서 적어도 5 분 동안 UDCA, tUDCA 및 gUDCA를 포함하는 UDCA 계열의 담즙산의 농도의 합이 UDCA 계열이 아닌 담즙산의 농도의 합보다 크게 될 수 있다.According to one embodiment of the present application, when the pharmaceutical composition is orally administered to a subject, the sum of the concentrations of UDCA-based bile acids including UDCA, tUDCA, and gUDCA for at least 5 minutes in the lung is equal to the concentration of non-UDCA-based bile acids. can be greater than the sum.
본원의 일 실시예에 의하면, 상기 UDCA 계열의 담즙산은 UDCA이고, 약학 조성물을 대상에 경구투여시 폐(lung) 조직 내에서 1시간 이내에 UDCA 농도가 0.65 ± 1.03 ㎍/g tissue 이상일 수 있다.According to an embodiment of the present application, the UDCA-based bile acid is UDCA, and when the pharmaceutical composition is orally administered to a subject, the UDCA concentration within 1 hour in lung tissue may be 0.65 ± 1.03 μg/g tissue or more.
본원의 일 실시예에 의하면, 상기 UDCA 계열의 담즙산은 tUDCA이고, 약학 조성물을 대상에 경구투여시 폐(lung) 조직 내에서 2시간 이내에 tUDCA 농도가 0.13 ± 0.17 ㎍/g tissue 이상일 수 있다.According to an embodiment of the present application, the UDCA-based bile acid is tUDCA, and when the pharmaceutical composition is orally administered to a subject, the tUDCA concentration within 2 hours in lung tissue may be 0.13 ± 0.17 μg/g tissue or more.
본원의 일 실시예에 의하면, 상기 제1 제제는 액상, 시럽, 또는 건조된 제제 형태일 수 있다.According to an embodiment of the present application, the first formulation may be in the form of a liquid, syrup, or dried formulation.
본원의 일 실시예에 의하면, 약학 조성물은 비강 또는 정맥 주사로 투여될 수 있다.According to one embodiment of the present application, the pharmaceutical composition may be administered by nasal or intravenous injection.
본원의 일 실시예에 의하면, 상기 제2 제제는 스테로이드 제제 또는 이의 약학적으로 허용가능한 염을 포함하고, 생존율을 증가시킬 수 있다.According to an embodiment of the present application, the second agent may include a steroid agent or a pharmaceutically acceptable salt thereof, and may increase the survival rate.
본원의 일 실시예에 의하면, 상기 제2 제제는 당질코르티코이드계일 수 있다.According to one embodiment of the present application, the second agent may be a glucocorticoid type.
본원의 일 실시예에 의하면, 상기 제2 제제는 1일 0.05 내지 10mg/kg의 치료학적 유효량으로 투여되도록 포함될 수 있다.According to an embodiment of the present application, the second agent may be included to be administered in a therapeutically effective amount of 0.05 to 10 mg/kg per day.
본원의 일 실시예에 의하면, 제1 제제 및 제2 제제는 서로 독립적으로 투여될 수 있다.According to an embodiment of the present application, the first agent and the second agent may be administered independently of each other.
본 발명은 수가용화된 우르소데옥시콜산(UDCA) 계열 담즙산을 포함하여, LPS 유발 패혈증 마우스 동물모델에서 사이토카인 폭풍을 일으키는 염증인자인 TNF-alpha, IL-6 의 과다 생성을 억제시켜 패혈증을 예방 또는 치료할 수 있다.The present invention prevents sepsis by inhibiting excessive production of inflammatory factors TNF-alpha and IL-6, which are inflammatory factors that cause a cytokine storm in an LPS-induced sepsis mouse animal model, including water-soluble ursodeoxycholic acid (UDCA)-based bile acids Or it can be treated.
본 발명은 수가용화된 담즙산 및 항염증 제제 또는 항알러지 제제를 포함하여 폐 조직 내 흡수도를 크게 높이고, 사이토카인 발현을 억제하고 생존율을 개선할 수 있는 박테리아, 호흡기 바이러스 등의 감염으로 인한 패혈증, 급성 호흡곤란증후군 또는 폐손상을 예방, 개선 및 치료할 수 있다.The present invention includes a water-solubilized bile acid and an anti-inflammatory or anti-allergic agent to significantly increase absorption in lung tissue, suppress cytokine expression, and improve survival rate of sepsis caused by infections such as bacteria and respiratory viruses; It can prevent, improve and treat acute respiratory distress syndrome or lung damage.
본 발명에 따른 수가용화된 우르소데옥시콜산(UDCA) 계열 담즙산은 LPS 유발 패혈증 마우스 동물모델에서 사이토카인 폭풍을 일으키는 염증인자인 TNF-alpha, IL-6 의 과다 생성을 억제시키고, 특히 덱사메타손을 저용량으로 병용해서 반복 경구투여 시 생존율까지 증가시킴으로써 패혈증, 급성 폐손상 및 급성 호흡곤란증 후군 치료에 유용하게 사용될 수 있다. The water-solubilized ursodeoxycholic acid (UDCA)-based bile acid according to the present invention inhibits the excessive production of TNF-alpha and IL-6, which are inflammatory factors that cause cytokine storm, in an LPS-induced sepsis mouse animal model. It can be usefully used in the treatment of sepsis, acute lung injury and acute respiratory distress syndrome by increasing the survival rate during repeated oral administration.
본 발명에 따른 수가용화된 우르소데옥시콜산(UDCA) 청정 수용액은 경구투여만으로도 폐 조직내로 UDCA를 유효적 치료량으로 신속히 전달시킬 수 있고, LPS- 유발 패혈증 동물모델에서 스테로이드 제제의 덱사메타손을 저용량으로 병용 반복 투여 시에는 생존율 및 폐질환을 개선시키는 것이 본 발명에 의하여 밝혀졌으므로 패혈증, 급성 폐손상 및 급성 호흡곤란증후군 치료에 유용하게 사용될 수 있다. The water-solubilized clean aqueous solution of ursodeoxycholic acid (UDCA) according to the present invention can rapidly deliver an effective therapeutic amount of UDCA into the lung tissue only by oral administration, and in an animal model of LPS-induced sepsis, dexamethasone of a steroid preparation is used in combination with a low dose. Since it was found by the present invention to improve the survival rate and lung disease during repeated administration, it can be usefully used in the treatment of sepsis, acute lung injury and acute respiratory distress syndrome.
본 발명의 일 실시예에 따르면, 수가용화된 UDCA, UDCA의 생체내 대사물인 tUDCA(taurodeoxycholic acid) 또는 gUDCA(glycodeoxycholic acid)를 폐 조직내까지 이상반응 없이 치료적 유효량으로 신속히 전달할 수 있는 패혈증, 급성 폐손상 질환, 및 급성 호흡곤란증후군 예방 또는 치료용 조성물을 제공할 수 있다.According to an embodiment of the present invention, sepsis, acute in which water-soluble UDCA, tUDCA (taurodeoxycholic acid) or gUDCA (glycodeoxycholic acid), which are in vivo metabolites of UDCA, can be rapidly delivered in a therapeutically effective amount to the lung tissue without adverse reactions A composition for preventing or treating lung injury disease and acute respiratory distress syndrome may be provided.
본 발명의 일 실시예에 따르면, 본 발명에 따른 조성물은 경구투여, 비강 투여, 또는 정맥주사로도 패혈증, 급성 폐손상 및 급성 호흡곤란증 질환 예방 또는 치료효과를 달성할 수 있고, 특히 경구투여인 경우 패혈증, 급성 폐손상 질환, 및 급성 호흡곤란증후군의 치료에 대한 환자의 편의성을 극대화시킬 수 있다는 이점이 있다.According to one embodiment of the present invention, the composition according to the present invention can achieve the prevention or treatment effect of sepsis, acute lung injury, and acute respiratory distress disease even by oral administration, nasal administration, or intravenous administration, and in particular, oral administration In this case, there is an advantage in that it is possible to maximize the convenience of the patient for the treatment of sepsis, acute lung injury disease, and acute respiratory distress syndrome.
따라서, 본 발명으로 조성물 중 유효성분인 UDCA 계열 담즙산 및 덱사메타손은 합성 의약품이므로 제조단가가 낮아 저렴한 비용만으로도 패혈증, 급성 폐손상 질환 및 급성 호흡곤란증후군의 효과적인 예방 또는 치료가 가능하다.Therefore, since the UDCA-based bile acid and dexamethasone, which are active ingredients in the composition of the present invention, are synthetic drugs, the manufacturing cost is low, and thus, it is possible to effectively prevent or treat sepsis, acute lung injury disease, and acute respiratory distress syndrome at a low cost.
도 1은 본 발명의 실시예 1-3에 의해 제조한 UDCA 용액의 pH값에 따른 청정 수용액 생성여부를 나타낸 것이다.1 shows whether a clean aqueous solution is generated according to the pH value of the UDCA solution prepared according to Example 1-3 of the present invention.
도 2는 본 발명의 실시예 1-4에 의해 제조한 UDCA 용액의 pH값에 따른 청정 수용액 생성여부를 나타낸 것이다.2 shows whether a clean aqueous solution is generated according to the pH value of the UDCA solution prepared according to Examples 1-4 of the present invention.
도 3은 본 발명의 실시예 1-5에 의해 제조한 UDCA 용액의 pH값에 따른 청정 수용액 생성여부를 나타낸 것이다.3 shows whether a clean aqueous solution is generated according to the pH value of the UDCA solution prepared according to Examples 1-5 of the present invention.
도 4는 본 발명의 실시예 1-6에 의해 제조한 UDCA 용액의 pH값에 따른 청정 수용액 생성 여부를 나타낸 것이다.4 shows whether a clean aqueous solution is generated according to the pH value of the UDCA solution prepared according to Examples 1-6 of the present invention.
도 5는 본 발명의 실시예 1-7에 의해 제조한 UDCA 용액의 pH값에 따른 청정 수용액 생성 여부를 나타낸 것이다.5 shows whether a clean aqueous solution is generated according to the pH value of the UDCA solution prepared according to Examples 1-7 of the present invention.
도 6은 본 발명의 실시예 1-8에 의해 제조한 UDCA 용액의 pH값에 따른 청정 수용액 생성 여부를 나타낸 것이다.6 shows whether a clean aqueous solution is generated according to the pH value of the UDCA solution prepared according to Examples 1-8 of the present invention.
도 7은 LPS 유발 패혈증 동물모델에서 혈청 내 초기 염증유발 사이토카인 TNF-alpha(cytokine TNF-alpha) 측정 결과를 시간에 따라 나타낸 것이다.7 is a graph showing the measurement results of the initial inflammation-inducing cytokine TNF-alpha (cytokine TNF-alpha) in serum in an LPS-induced sepsis animal model over time.
도 8은 LPS 유발 패혈증 동물모델에서 혈청 내 초기 염증유발 사이토카인 IL-6(cytokine IL-6) 측정 결과를 시간에 따라 나타낸 것이다.8 is a graph showing the measurement results of the initial inflammation-inducing cytokine IL-6 (cytokine IL-6) in serum in an LPS-induced sepsis animal model over time.
도 9는 LPS 유발 패혈증 동물모델에서 UDCA 정제 및 수가용화 UDCA 청정 수용액의 단독 반복투여 시의 생존율을 나타낸 것이다.Figure 9 shows the survival rate in the case of single repeated administration of UDCA purification and aqueous solubilized UDCA clean aqueous solution in an LPS-induced sepsis animal model.
도 10은 LPS 유발 패혈증 동물모델에서 덱사메타손의 용량에 따른 단독 반복투여 시의 생존율 나타낸 것이다.10 is a graph showing the survival rate in case of repeated administration alone according to the dose of dexamethasone in an LPS-induced sepsis animal model.
도 11은 LPS 유발 패혈증 동물모델에서 수가용화 UDCA 청정수용액과 저용량의 덱사메타손을 병용 반복 투여에 따른 생존율을 나타낸 것이다. 11 shows the survival rate according to repeated administration of water-soluble UDCA clean aqueous solution and low-dose dexamethasone in combination in an LPS-induced sepsis animal model.
도 12는 본 발명에 의한 수가용화된 UDCA 청정 수용액 제제인 AGP600의 경구투여 후 폐 조직내까지 운반된 UDCA의 시간에 따른 변화량을 보여주는 약동학(PK) 데이터이다.12 is pharmacokinetic (PK) data showing the change with time of UDCA delivered to the lung tissue after oral administration of AGP600, which is a water-solubilized UDCA clean aqueous solution formulation according to the present invention.
도 13은 본 발명에 의한 수가용화된 UDCA 청정 수용액 제제인 AGP600의 경구투여 후 폐 조직내까지 운반된 UDCA 계열 담즙산들(UDCA, TUDCA, GUDCA 같은 UDCA type bile acids) 및 비UDCA 계열 담즙산들의 시간에 따른 변화량을 보여주는 약동학(PK) 데이터이다.13 shows UDCA-type bile acids (UDCA-type bile acids such as UDCA, TUDCA, and GUDCA) and non-UDCA-type bile acids delivered to the lung tissue after oral administration of AGP600, a water-solubilized UDCA aqueous solution formulation according to the present invention. Pharmacokinetic (PK) data showing the amount of change.
도 14는 본 발명에 의한 수가용화된 UDCA 청정 수용액 제제인 AGP600의 경구투여 후 폐 조직내까지 운반된 UDCA 및 UDCA의 대사물, 즉 UDCA, tUDCA 및 gUDCA의 농도를 합한 것(UDCA 계열 담즙산)과 기타 담즙산들(비UDCA 계열 담즙산)의 농도를 합한 것의 시간에 따른 변화량을 보여주는 약동학(PK) 데이터이다.14 shows the sum of the concentrations of UDCA and UDCA metabolites, i.e., UDCA, tUDCA and gUDCA, delivered to the lung tissue after oral administration of AGP600, a water-solubilized UDCA clean aqueous solution formulation according to the present invention (UDCA-based bile acids); Pharmacokinetic (PK) data showing the change over time of the sum of the concentrations of other bile acids (non-UDCA bile acids).
도 15는 본 발명에 의한 수가용화 UDCA 계열 담즙산을 포함하는 AGP600의 경구투여 후 폐 조직내까지 운반된 담즙산들의 시간에 따른 변화량을 보여주는 약동학(PK) 데이터이다.15 is pharmacokinetic (PK) data showing changes with time of bile acids transported to the lung tissue after oral administration of AGP600 containing a water-solubilized UDCA-based bile acid according to the present invention.
본 발명을 더 쉽게 이해하기 위해 편의상 특정 용어를 본원에 정의한다. 본원에서 달리 정의하지 않는 한, 본 발명에 사용된 과학 용어 및 기술 용어들은 해당 기술 분야에서 통상의 지식을 가진 자에 의해 일반적으로 이해되는 의미를 가질 것이다. 또한, 문맥상 특별히 지정하지 않는 한, 단수 형태의 용어는 그것의 복수 형태도 포함하는 것이며, 복수 형태의 용어는 그것의 단수 형태도 포함하는 것으로 이해되어야 한다. In order to better understand the present invention, certain terms are defined herein for convenience. Unless defined otherwise herein, scientific and technical terms used herein shall have the meanings commonly understood by one of ordinary skill in the art. Also, unless the context specifically dictates otherwise, it is to be understood that a term in the singular includes its plural form as well, and a term in the plural form also includes its singular form.
본 발명에서 사용되는 용어 "치료"는 본 발명의 조성물의 투여로 패혈증, 급성 폐손상 또는 급성 호흡 곤란증 질환의 증상이 악화되지 않거나 호전 또는 이롭게 변경되는 모든 행위를 의미한다. As used herein, the term “treatment” refers to any action in which the symptoms of sepsis, acute lung injury, or acute respiratory distress disease do not worsen, improve, or change advantageously by administration of the composition of the present invention.
본 발명에서 사용되는 용어 "유효성분으로 포함하는" 또는 "치료적 활성량으로 포함하는"의 의미는 패혈증, 급성 폐손상 질환 또는 급성 호흡곤란증후군의 예방 또는 치료용 조성물, 경구투여제 및 정맥주사제로써 효과를 나타낼 수 있는 정도, 예를 들어, 예방효과, 치료효과 등을 나타낼 수 있을 정도로 함유하는 것을 의미한다.As used herein, the term “comprising as an active ingredient” or “comprising a therapeutically active amount” means a composition for preventing or treating sepsis, acute lung injury disease, or acute respiratory distress syndrome, oral administration, and intravenous injection As such, it means to contain enough to exhibit an effect, for example, a preventive effect, a therapeutic effect, and the like.
본 발명에서 사용되는 용어 "예방"은 패혈증, 급성 폐손상 또는 급성 호흡곤란증 질환 발병 전부터 약물을 처치하여, 발병 후에 경구복용을 시킨 후 치료되는 상태와 관련된 파라미터, 예를 들면 증상의 정도를 적어도 감소시키는 모든 행위를 의미한다.The term "prevention" as used in the present invention is to treat the drug before the onset of sepsis, acute lung injury or acute respiratory distress disease, and then take oral administration after the onset of the disease, and then at least reduce the parameters related to the condition to be treated, for example, the severity of symptoms. It means any action you do.
본 발명에서 사용되는 용어 "투명한(clear) 수용액" 또는 "청정 수용액"은 육안상 침전물이 실질적으로 없는 용액 상태를 의미한다.As used herein, the term "clear aqueous solution" or "clean aqueous solution" refers to a solution state substantially free of deposits visually.
본 발명에서 사용되는 용어 "패혈증 또는 급성 폐손상 질환"은 패혈증, 급성 폐손상 질환, 또는 급성 호흡곤란증후군을 포함한다.As used herein, the term "septicemia or acute lung injury disease" includes sepsis, acute lung injury disease, or acute respiratory distress syndrome.
본 발명에서 사용되는 용어 "YSB201, YSB201-xx 혹은 AGP600"은 수가용화된 UDCA 청정 수용액 제제를 지칭하는 코드명이다.As used herein, the term "YSB201, YSB201-xx or AGP600" is a code name for a water-solubilized UDCA clean aqueous solution formulation.
본 발명에서 사용되는 용어 "우르소데옥시콜산(Ursodeoxycholic acid, UDCA) 계열 담즙산"은 우르소데옥시콜산(ursodeoxycholic acid, UDCA), 타우로우르소데옥시콜산(tauro-ursodeoxycholic acid, tUDCA) 및 글리코우르소데옥시콜산(glyco-ursodeoxycholic acid, gUDCA)를 포함한다.As used herein, the term "ursodeoxycholic acid (UDCA)-based bile acid" refers to ursodeoxycholic acid (UDCA), tauro-ursodeoxycholic acid (tUDCA) and glycoursode. Contains glyco-ursodeoxycholic acid (gUDCA).
본 발명자들은 내독소 (Lipopolysaccharide; LPS) 유발 패혈증 마우스 동물모델에서 수가용화된 UDCA 계열 담즙산을 투여한 후 사이토카인 폭풍을 일으키는 염증인자인 TNF-alpha, IL-6 의 과다 생성 억제 효과와 특히 항염증 제제 또는 항알러지 제제를 저용량으로 병용해서 반복 투여 시 생존율 증가를 확인함으로써, 본 발명을 완성하게 되었다. The present inventors have investigated the effect of inhibiting the excessive production of TNF-alpha and IL-6, which are inflammatory factors that cause cytokine storm, after administration of water-solubilized UDCA-based bile acids in a mouse animal model of endotoxin (Lipopolysaccharide; LPS)-induced sepsis, and in particular, anti-inflammatory The present invention was completed by confirming an increase in survival rate upon repeated administration by using a low dose of an agent or an anti-allergic agent in combination.
본원의 일 측면에 의하면, 수가용화된 우르소데옥시콜산(Ursodeoxycholic acid, UDCA) 계열 담즙산 또는 이의 약학적으로 허용가능한 염을 포함하는 제1 제제; 및 치료학적으로 허용가능한 항염증 제제 또는 항알러지 제제인 제2 제제;를 포함하고, 염증성 사이토카인 발현을 억제하는, 패혈증 예방 또는 치료용, 약학 조성물이 제공된다.According to one aspect of the present application, a first formulation comprising a water-solubilized Ursodeoxycholic acid (UDCA)-based bile acid or a pharmaceutically acceptable salt thereof; And a therapeutically acceptable anti-inflammatory agent or an anti-allergic agent, a second agent; including, and inhibiting the expression of inflammatory cytokines, sepsis prevention or treatment, a pharmaceutical composition is provided.
본원의 다른 측면에 의하면, 수가용화된 우르소데옥시콜산(Ursodeoxycholic acid, UDCA) 계열 담즙산 또는 이의 약학적으로 허용가능한 염을 포함하는 제1 제제; 및 치료학적으로 허용가능한 항염증 제제 또는 항알러지 제제인 제2 제제;를 포함하고, 급성 폐손상 질환 또는 급성 호흡곤란증후군 예방 또는 치료용, 약학 조성물이 제공된다.According to another aspect of the present application, a first agent comprising a water-solubilized Ursodeoxycholic acid (UDCA) series bile acid or a pharmaceutically acceptable salt thereof; And a therapeutically acceptable anti-inflammatory agent or an anti-allergic agent, a second agent; including, for the prevention or treatment of acute lung injury disease or acute respiratory distress syndrome, a pharmaceutical composition is provided.
본원의 약학 조성물은 제1 제제에 수가용화된 우르소데옥시콜산(Ursodeoxycholic acid, UDCA) 계열 담즙산을 포함하는 것을 특징으로 한다.The pharmaceutical composition of the present application is characterized in that it comprises a ursodeoxycholic acid (UDCA) series bile acid solubilized in the first agent.
본원에 있어서, 상기 UDCA 계열 담즙산은 가용성 UDCA, 수가용성 UDCA 유도체, UDCA 염, 및 아민과 컨쥬게이트된 UDCA에서 선택되는 UDCA로서 수가용화된 것일 수 있다. 또한, 이에 한정하는 것은 아니나, UDCA의 수가용성 금속염 및 수가용성 O-설폰화 담즙산도 가용성 UDCA 염에 포함될 수 있다. In the present application, the UDCA-based bile acid may be a UDCA selected from soluble UDCA, water-soluble UDCA derivatives, UDCA salts, and UDCA conjugated with amines, and may be water-solubilized. In addition, although not limited thereto, water-soluble metal salts and water-soluble O-sulfonated bile acids of UDCA may also be included in the soluble UDCA salts.
본원에 있어서, 상기 수가용화된 UDCA 계열 담즙산은 수가용화된 우르소데옥시콜산(UDCA), 타우로우르소데옥시콜산(tUDCA) 및 글리코우르소데옥시콜산(gUDCA) 중에서 선택되는 1종 이상을 포함할 수 있다. 상기 타우로우르소데옥시콜산(tUDCA) 및 글리코우르소데옥시콜산(gUDCA)은 UDCA의 생체 내 대사물 즉 유도체로, tUDCA는 타우린과 컨쥬게이트된 UDCA 유도체이고, gUDCA는 글라이신과 컨쥬게이트된 우르소데옥시콜산 유도체를 의미한다.In the present application, the water-solubilized UDCA-based bile acid may include at least one selected from water-solubilized ursodeoxycholic acid (UDCA), tauroursodeoxycholic acid (tUDCA), and glycoursodeoxycholic acid (gUDCA). can The tauroursodeoxycholic acid (tUDCA) and glycoursodeoxycholic acid (gUDCA) are in vivo metabolites or derivatives of UDCA, tUDCA is a UDCA derivative conjugated with taurine, and gUDCA is ursode conjugated with glycine. oxycholic acid derivatives.
우르소데옥시콜산(UDCA)은 친수성 담즙산이고, 경구투여가 가능하며 인체 내 총 담즙산의 단지 약 3% 정도의 낮은 농도이지만, 통상 인간의 담즙에 존재하며, 미국 FDA에 의해 원발성 담즙정체성 간경변증 치료제로서 승인된 유일한 약물이다. UDCA의 약리 작용은 항산화작용, 항염증작용, 항세포사멸억제작용 등이 있다. 이런 효과들은 폐 질환의 치료에 아주 중요한 기작이며, UDCA가 단일분자로 역할을 할 때 더 극명하게 나타난다. 따라서 이런 효과가 있는 물질을 어떻게 인체의 폐 조직내까지 잘 흡수 및 전달할 수 있느냐가 문제이다. 결정형 UDCA가 피부 발적물로 분류되어 있고, 친수성과 친유성을 동시에 가지는 양매성 분자이어서 물에 거의 녹지 않고 소량 녹더라도 이량체(dimers), 사량체(tetramers), 또는 미셀(micell) 형태를 띠므로 UDCA 단일분자로서 작용하기 어렵기 때문에, 본 발명의 조성물은 UDCA 결정체를 물에 고농도로 가용화시켜 단일분자로서 기능을 나타낼 수 있게 하였다. Ursodeoxycholic acid (UDCA) is a hydrophilic bile acid, can be administered orally, and has a low concentration of only about 3% of total bile acids in the human body, but is usually present in human bile, It is the only drug approved. The pharmacological actions of UDCA include antioxidant action, anti-inflammatory action, and anti-apoptotic action. These effects are a very important mechanism for the treatment of lung disease, and are more pronounced when UDCA acts as a single molecule. Therefore, the problem is how to absorb and deliver a substance having such an effect well into the lung tissue of the human body. Crystalline UDCA is classified as a skin redness and is an amphipathic molecule having both hydrophilicity and lipophilicity. Therefore, since it is difficult to act as a single molecule of UDCA, the composition of the present invention enables the UDCA crystals to function as a single molecule by solubilizing them in water at a high concentration.
종래의 결정형 UDCA 제제형태인 정제나 캡슐제제로 경구투여 시, 전체 투여량 대비 약 30 ~ 60%는 비이온 수동 확산에 의해 빈창자 및 돌창자를 따라 흡수되고, 결정형 UDCA는 불용성이기 때문에 결장에서 소량(섭취 용량의 20%)만이 능동 수송 기전에 의해 돌창자에 흡수된다. UDCA가 간세포에 의해 흡수되면, tUDCA 및 gUDCA로 컨쥬게이션 될 수 있고, UDCA와 결합된 tUDCA 및 gUDCA는 인간에서 분비된 담즙산으로 간의 일차 통과 청소율에 의해 배설되므로 경구투여 후 UDCA의 혈액 내 농도는 매우 낮게 된다. 따라서, 기존 UDCA 제제로 경구투여 시 폐 조직내까지 UDCA를 운반할 수 있다는 연구결과가 발표된 적이 없으며, 특히 패혈증 또는 급성 폐손상과 같은 패혈증 또는 급성 폐손상 질환의 예방 또는 치료용 조성물로 제공된 예가 없다. 이 문제를 해결하기 위해서는, 패혈증, 급성 폐손상 질환, 및 급성 호흡곤란증후군을 치료하기 위해서는 최우선적으로 폐 조직까지 UDCA 계열 담즙산을 치료적으로 유효적인 농도로 효율적으로 운반시켜 줘야 한다. When administered orally in tablets or capsules, which are conventional crystalline UDCA formulations, about 30 to 60% of the total dose is absorbed along the stomach and intestines by nonionic passive diffusion. Only (20% of the ingested dose) is absorbed by the active transport mechanism into the ileus. When UDCA is absorbed by hepatocytes, it can be conjugated to tUDCA and gUDCA, and tUDCA and gUDCA bound to UDCA are excreted as bile acids secreted in humans by the liver's first-pass clearance, so the blood concentration of UDCA after oral administration is very high. goes low Therefore, no research results have been published that UDCA can be transported into the lung tissue when orally administered with an existing UDCA formulation. none. In order to solve this problem, in order to treat sepsis, acute lung injury, and acute respiratory distress syndrome, UDCA-based bile acids must be efficiently transported to the lung tissue at a therapeutically effective concentration.
본원의 상기 구성에 의해, 그 화학적 특성상 장-간 순환물질(enterohepatic circulation)로 분류되어 있고 경구투여 시 간의 일차통과청소율 (hepatic first-pass clearance)이 높기 때문에 폐 조직내까지 UDCA 계열 담즙산을 고농도로 운반할 수 없었던 결정형 UDCA 계열 담즙산의 근본적인 문제점을 해결할 수 있다. According to the composition of the present application, because of its chemical characteristics, it is classified as enterohepatic circulation and the hepatic first-pass clearance during oral administration is high. It is possible to solve the fundamental problem of crystalline UDCA-based bile acids that could not be transported.
즉, 본원의 약학 조성물은 경구투여만으로도 UDCA 계열 담즙산이 폐 조직내까지 빠른 시간내 치료적 활성량이 운반될 수 있도록, UDCA 계열 담즙산을 수가용화 시켜 청정수용액으로 제제화 하여, 패혈증, 급성 폐손상 질환 또는 급성 호흡곤란증후군의 예방 또는 치료용 조성물로 제공할 수 있다.That is, the pharmaceutical composition of the present application is formulated into a clean aqueous solution by dissolving UDCA-based bile acids so that the therapeutically active amount of the UDCA-based bile acids can be transported to the lung tissue within a short period of time by oral administration alone, It can be provided as a composition for preventing or treating acute respiratory distress syndrome.
이에 한정되는 것은 아니나, 본원의 약학 조성물 내 UDCA의 용해도가 결정형 UDCA의 물에 대한 용해도의 약 3,000배 이상(0.15 M 대 0.05 mM) 일 수 있고, tUDCA와 비교하여, 약 300배 이상(0.15 M 대 0.45 mM)일 수 있다. 따라서, 본 출원인은 수가용화된 UDCA 계열 담즙산을 이용하여 패혈증, 급성 폐손상 질환 또는 급성 호흡곤란증후군 치료용 약학 조성물에 관한 본 발명을 완성하였다.Although not limited thereto, the solubility of UDCA in the pharmaceutical composition of the present application may be about 3,000 times or more (0.15 M vs. 0.05 mM) of the solubility of crystalline UDCA in water, and compared to tUDCA, about 300 times or more (0.15 M) vs. 0.45 mM). Accordingly, the present applicant has completed the present invention regarding a pharmaceutical composition for the treatment of sepsis, acute lung injury, or acute respiratory distress syndrome using water-solubilized UDCA-based bile acids.
따라서 본원의 약학 조성물을 마우스에 경구투여 시(125mg/kg) PK 분석(pharmacokinetics study) 결과, 종래의 결정형 UDCA 제형(정제, 캡슐제)에서는 나타나지 않았던 폐 조직 내 UDCA 최고혈중농도 7.13 ± 2.29 ㎍/g tissue와 Tmax 5분의 특징이 나타났다. 즉, 기존 제제에서는 달성할 수 없었던 경구투여임에도 폐 조직내로 고농도 UDCA를 운반시킬 수 있고 패혈증 또는 급성 폐손상 질환의 예방 또는 치료효과를 달성할 수 있다.Therefore, as a result of PK analysis (pharmacokinetics study) when the pharmaceutical composition of the present application is orally administered to a mouse (125 mg/kg), the UDCA peak plasma concentration in the lung tissue, which did not appear in the conventional crystalline UDCA formulation (tablet, capsule), 7.13 ± 2.29 μg/ Characteristics of g tissue and Tmax of 5 min were shown. That is, it is possible to deliver a high concentration of UDCA into the lung tissue despite oral administration, which could not be achieved with existing formulations, and to achieve the prevention or treatment effect of sepsis or acute lung injury.
본원의 약학 조성물은, 상기와 같이 UDCA 계열 담즙산을 폐 조직까지 신속하게 치료적 유효량으로 운반하여, 사이토카인 폭풍의 원인물질인 초기 염증유발인자 TNF-alpha와 IL-6 의 과다 발현을 억제시킬 수 있다. 본원의 약학 조성물은 단백질의 발현을 유전자 수준에서 하향조절할 수 있다The pharmaceutical composition of the present application, as described above, rapidly transports the UDCA-based bile acids to the lung tissue in a therapeutically effective amount, thereby inhibiting the overexpression of the initial inflammatory factors TNF-alpha and IL-6, which are the causative agents of the cytokine storm. there is. The pharmaceutical composition of the present disclosure may down-regulate the expression of a protein at the gene level.
나아가, 본원의 약학 조성물은, 치료학적으로 허용가능한 항염증 제제인 제2 제제;를 조합하여 사이토카인 발현 억제뿐만 아니라 생존율까지 증가시킬 수 있다. 또한, 상기와 같은 제1 제제 및 제2 제제의 조합에 의해, 제2 제제인 항염증 제제의 투여량을 줄일 수 있어 항염증 제제에 의한 부작용을 최소화할 수 있다.Furthermore, the pharmaceutical composition of the present application may increase the survival rate as well as inhibition of cytokine expression by combining the second agent, which is a therapeutically acceptable anti-inflammatory agent. In addition, by the combination of the first agent and the second agent as described above, it is possible to reduce the dosage of the second agent, the anti-inflammatory agent, it is possible to minimize the side effects caused by the anti-inflammatory agent.
이에 한정되는 것은 아니나, 상기 제2 제제는 스테로이드 제제 또는 이의 약학적으로 허용가능한 염을 포함할 수 있다. 상기 구성에 의하면, 제2 제제인 스테로이드 제제의 투여량을 줄일 수 있어 스테로이드 제제에 의한 부작용을 최소화할 수 있다. Although not limited thereto, the second agent may include a steroid agent or a pharmaceutically acceptable salt thereof. According to the above configuration, it is possible to reduce the dosage of the steroid preparation, which is the second preparation, so that the side effects caused by the steroid preparation can be minimized.
부신피질호르몬제의 스테로이드는 고용량 장기간 사용시 부작용으로는 감염증 유발이나 감염증 악화가 나타날 수 있으며, 부신피질 기능부전, 당뇨병, 쿠싱증후군, 소화성 궤양, 위통, 가슴쓰림, 우울증, 불면, 골다공증, 근육통, 지방간, 부종, 고혈압, 체액저류, 녹내장, 백내장, 시야흐림, 혈전증, 여드름, 색소 침착, 얇고 연약한 피부, 발진, 체중증가 등을 유발할 수 있다. Adrenal corticosteroid steroids can cause infections or worsen infections as side effects when used in high doses for a long time, and adrenocortical dysfunction, diabetes, Cushing's syndrome, peptic ulcer, stomach pain, heartburn, depression, insomnia, osteoporosis, muscle pain, fatty liver , edema, high blood pressure, fluid retention, glaucoma, cataracts, blurred vision, thrombosis, acne, pigmentation, thin and fragile skin, rash, and weight gain.
따라서, 패혈증이나 급성 호흡곤란증 같은 치료에, 특히 중증 코로나바이러스감염증-19(covid-19) 환자에서도 고용량을 장기간 사용하지 못하고 저용량 단기간 처방에만 그치고 있다. 특히 의학 전문가들은 "사이토카인 폭풍 등 염증성 질환이 발생했을 때 코르티코이스테로이드를 투여해 급한 면역세포의 활동을 막아 염증을 완화하는 것을 맞지만, 장기적으로 투여할 경우 면역이 약해지면서 오히려 코로나바이러스감염증-19에 악영향을 줄 수 있기 때문에 주의가 필요하다는 그런 의견을 주고 있다"고 밝혔다.Therefore, treatment such as sepsis or acute respiratory distress, especially in patients with severe coronavirus infection-19 (covid-19), cannot use high doses for a long period of time, and only low doses are prescribed for a short period of time. In particular, medical experts said, "When an inflammatory disease such as a cytokine storm occurs, it is correct to administer corticosteroids to block the activity of urgent immune cells to relieve inflammation. We are giving the opinion that caution is necessary because it can have an adverse effect,” he said.
이에 한정되는 것은 아니나, 상기 제2 제제는 당질코르티코이드계일 수 있다. Although not limited thereto, the second agent may be a glucocorticoid-based agent.
이에 한정되는 것은 아니나, 본원에서 스테로이드 제제는 덱사메타손 또는 이의 약학적으로 허용가능한 염, 히드로코르티손(hydrocortisone) 또는 이의 약학적으로 허용가능한 염, 코르티손(cortisone) 또는 이의 약학적으로 허용가능한 염, 프레드니손(prednisone) 또는 이의 약학적으로 허용가능한 염, 프레드니솔론(prednisolone) 또는 이의 약학적으로 허용가능한 염, 메틸프레드니솔론( methylprednisolone) 또는 이의 약학적으로 허용가능한 염, 트리암시놀론(triamcinolone) 또는 이의 약학적으로 허용가능한 염, 베타메타손(betamethasone) 또는 이의 약학적으로 허용가능한 염으로 이루어진 군으로부터 선택될 수 있다. 이에 한정되는 것은 아니나, 상기 스테로이드 제제는 덱사메타손 또는 이의 약학적으로 허용가능한 염이 적합할 수 있다. Although not limited thereto, the steroid preparation herein includes dexamethasone or a pharmaceutically acceptable salt thereof, hydrocortisone or a pharmaceutically acceptable salt thereof, cortisone or a pharmaceutically acceptable salt thereof, prednisone ( prednisone) or a pharmaceutically acceptable salt thereof, prednisolone or a pharmaceutically acceptable salt thereof, methylprednisolone or a pharmaceutically acceptable salt thereof, triamcinolone or a pharmaceutically acceptable salt thereof , may be selected from the group consisting of betamethasone or a pharmaceutically acceptable salt thereof. Although not limited thereto, dexamethasone or a pharmaceutically acceptable salt thereof may be suitable for the steroid preparation.
이에 한정되는 것은 아니나, 상기 제2 제제는 0.1 내지 10mg/kg/day의 치료학적 유효량으로 투여되도록 포함될 수 있다. 상기 제2 제제는 12세 이상·체중 최소 40kg 이상의 청소년 및 성인 환자들을 대상으로 경구복용하거나, 주사제로 투여하거나, 정맥을 통해 주입하는 방식으로 1일 1회 6mg, 최대 10일까지 치료학적 유효량으로 투여되는 것이 부작용을 최소화면서 패혈증, 급성 폐손상 질환, 또는 급성 호흡곤란증후군 치료면에서 적합할 수 있다. Although not limited thereto, the second agent may be included to be administered in a therapeutically effective amount of 0.1 to 10 mg/kg/day. The second formulation is administered orally, administered as an injection, or injected through a vein for adolescents and adults aged 12 years or older and weighing at least 40 kg. 6 mg once a day, a therapeutically effective amount for up to 10 days. Administration may be suitable for the treatment of sepsis, acute lung injury, or acute respiratory distress syndrome with minimal side effects.
상기 덱사메타손 또는 이의 약학적으로 허용가능한 염의 치료학적 유효량은 덱사메타손으로서 약 0.05 내지 약 1.5 mg/kg/day의 범위가 부작용을 최소화면서 패혈증, 급성 폐손상 질환, 또는 급성 호흡곤란증후군 치료면에서 적합할 수 있다. 일 구현예에서, 덱사메타손 또는 이의 약학적으로 허용가능한 염의 치료학적 유효량은 약 0.05 내지 약 1.0 mg/kg/day의 범위가 더 적합할 수 있고, 다른 구현예에서 치료학적 유효량은 0.05 내지 약 0.5 mg/kg/day의 범위가 더 적합할 수 있고, 또 다른 구현예에서 치료학적 유효량은 약 0.05 내지 약 0.25 mg/kg/day의 범위가 더욱더 적합할 수 있고, 또 다른 구현예에서 치료학적 유효량은 약 0.05 내지 약 0.15 mg/kg/day의 범위가 더욱더 적합할 수 있고, 약 0.05 내지 약 0.1mg/kg/day의 범위가 더욱더 적합할 수 있다. The therapeutically effective amount of dexamethasone or a pharmaceutically acceptable salt thereof is dexamethasone in the range of about 0.05 to about 1.5 mg/kg/day, while minimizing side effects, in terms of treatment of sepsis, acute lung injury, or acute respiratory distress syndrome. can In one embodiment, the therapeutically effective amount of dexamethasone or a pharmaceutically acceptable salt thereof may be more suitably in the range of about 0.05 to about 1.0 mg/kg/day, and in another embodiment, the therapeutically effective amount is 0.05 to about 0.5 mg A range of /kg/day may be more suitable, and in another embodiment a therapeutically effective amount may be more suitable in the range of about 0.05 to about 0.25 mg/kg/day, and in another embodiment a therapeutically effective amount is A range from about 0.05 to about 0.15 mg/kg/day may be even more suitable, and a range from about 0.05 to about 0.1 mg/kg/day may be even more suitable .
덱사메타손은 당질코르티코이드 (glucocorticoid) 계열의 합성 성분이고 항염증, 항알레르기, 항내독성 효력을 가지고 있어, 염증 및 자가면역 질환에 사용되고 있으나(Zhong-hua Wang et al., 2013), 투여할 때 염증억제를 과도하게 함으로써 오히려 면역반응을 저하시켜 세균이나 바이러스를 억제하지 못한다던가, 감염 및 위장관 출혈 등의 여러 부작용을 고려해야 한다. 특히, 부신피질호르몬제의 스테로이드는 고용량을 장기간 사용 시, 오히려 면역약화로 감염증 유발이나 감염증 악화 등의 부작용을 일으킬 수 있으므로, 부신피질 기능부전, 당뇨병, 쿠싱증후군, 소화성궤양, 위통, 가슴쓰림, 우울증, 불면, 골다공증, 근육통, 지방간, 부종, 고혈압, 체액저류, 녹내장, 백내장, 시야흐림, 혈전증, 여드름, 색소 침착, 얇고 연약한 피부, 발진, 체중 증가 등 다양한 부작용을 유발할 수 있다. 따라서, 덱사메타손 투여 시에는 상기 여러가지 부작용을 고려해야 하고, 덱사메타손은 아직까지 어느 국가에서도 패혈증 전용 치료제로 허가받지 못한 상태이다. 그러나, 본원의 약학 조성물은 상기와 같이 저용량을 투여하므로 상기 부작용으로 인한 문제점을 사전에 방지할 수 있다.Dexamethasone is a synthetic component of the glucocorticoid family and has anti-inflammatory, anti-allergic, and anti-endotoxic effects, so it is used for inflammatory and autoimmune diseases (Zhong-hua Wang et al., 2013), but when administered, it suppresses inflammation. By overdosing it, it is necessary to consider various side effects such as failure to suppress bacteria or viruses by lowering the immune response, infection, and gastrointestinal bleeding. In particular, when high-dose steroids are used for a long time, they can cause side effects such as infection or worsening of infections due to weakened immunity. It can cause various side effects such as depression, insomnia, osteoporosis, muscle pain, fatty liver, edema, high blood pressure, fluid retention, glaucoma, cataract, blurred vision, thrombosis, acne, pigmentation, thin and fragile skin, rash, and weight gain. Therefore, when administering dexamethasone, the various side effects should be considered, and dexamethasone has not yet been approved as a sepsis-only treatment in any country. However, since the pharmaceutical composition of the present application is administered at a low dose as described above, it is possible to prevent problems due to the side effects in advance.
본원의 약학적 조성물의 투여는 제1 제제인 수가용화된 UDCA 계열 담즙산의 투여와 제2 제제의 투여가 동일 시각에 수행될 수 있거나, 또는 이들 두 제제는 동일한 또는 중복되는 시간 주기(예를 들어, 동일 시간, 동일 일자 또는 동일한 주) 동안 간단히 투여될 수 있다. The administration of the pharmaceutical composition of the present disclosure may be performed at the same time as the administration of the first agent, the water-solubilized UDCA-based bile acid, and the second agent, or the two agents may be administered in the same or overlapping time period (eg, , at the same time, on the same day or during the same week).
또한, 본원의 약학 조성물에서 제1 제제 및 제2 제제는 제제형태 및/또는 투여시기면에서 서로 독립적으로 투여될 수 있다. 이에 한정되는 것은 아니나, 제1 제제는 경구투여되고, 제2 제제는 정맥/근육 주사로 투여될 수 있다. 제1 제제 및 제2 제제가 모두 경구투여되는 경우에도, 투여시기를 달리하여 투여될 수 있다. In addition, in the pharmaceutical composition of the present application, the first agent and the second agent may be administered independently of each other in terms of formulation form and/or administration timing. Although not limited thereto, the first agent may be administered orally, and the second agent may be administered by intravenous/muscular injection. Even when both the first agent and the second agent are orally administered, they may be administered with different administration timings.
이에 한정되는 것은 아니나, 본원에 있어서 급성 폐손상 질환 또는 급성 호흡곤란증후군은 세균 또는 바이러스 감염에 의한 것일 수 있다. Although not limited thereto, in the present application, the acute lung injury disease or acute respiratory distress syndrome may be caused by bacterial or viral infection.
이에 한정되는 것은 아니나, 본원에 있어서 상기 바이러스는 호흡기 바이러스일 수 있고, 호흡기 바이러스는 인플루엔자 바이러스(influenza virus), 사스 코로나바이러스(SARS-CoV), 메르스 코로나바이러스(MERS-CoV), COVID-19 바이러스(SARS-CoV-2), 아데노바이러스(adenovirus), 파라인플루엔자 바이러스 (parainfluenza virus, PIV), RS 바이러스(respiratory syncytial virus, RSV), 라이노바이러스(rhinovirus) 등일 수 있다. 특히, 본원에 있어서 상기 호흡기 바이러스는 사스코로나바이러스, 메르스 코로나바이러스, 또는 COVID-19 바이러스일 수 있다.Although not limited thereto, in the present application, the virus may be a respiratory virus, and the respiratory virus is an influenza virus, SARS-CoV, MERS-CoV, COVID-19. Virus (SARS-CoV-2), adenovirus (adenovirus), parainfluenza virus (PIV), RS virus (respiratory syncytial virus, RSV), rhinovirus (rhinovirus), etc. may be. In particular, in the present application, the respiratory virus may be a SARS coronavirus, a MERS coronavirus, or a COVID-19 virus.
이에 한정되는 것은 아니나, 본원에 있어서 상기 급성 폐손상 질환은 폐 염증 또는 폐 병변을 일으키고, 생존율을 감소시키는 것일 수 있다.Although not limited thereto, in the present application, the acute lung injury disease may be one that causes lung inflammation or lung lesions, and reduces the survival rate.
본원에 있어서, 본원의 약학 조성물은 치료학적 유효량의 UDCA 계열 담즙산 또는 이의 약학적으로 허용가능한 염을 투여하기에 적합한 투여 형태를 가질 수 있다.In the present application, the pharmaceutical composition of the present application may have a dosage form suitable for administering a therapeutically effective amount of a UDCA family bile acid or a pharmaceutically acceptable salt thereof.
이에 한정되는 것은 아니나, 본원의 약학 조성물은 경구투여용, 비강내 투여(intranasal administration) 또는 정맥주사용 제제 형태일 수 있다. Although not limited thereto, the pharmaceutical composition of the present application may be in the form of a formulation for oral administration, intranasal administration, or intravenous administration.
이에 한정되는 것은 아니나, 경구투여용 제제 형태가 패혈증, 급성 폐손상 질환 및 급성 호흡곤란증후군의 치료에 대한 환자의 편의성을 극대화시킬 수 있다는 점에서 더 적합할 수 있다.Although not limited thereto, the formulation form for oral administration may be more suitable in that it can maximize the convenience of the patient for the treatment of sepsis, acute lung injury disease, and acute respiratory distress syndrome.
본원에서는 상술한 바와 같이, 경구투여만으로도 UDCA 계열 담즙산의 폐 조직내 약동학 특성을 크게 향상시킨 수가용화된 UDCA 청정 수용액 제제를 포함하여 신속하게 패혈증, 급성 폐손상 질환 또는 급성 호흡곤란증후군의 예방 또는 치료할 수 있다. 즉, 본원의 약학 조성물을 경구투여만으로도 UDCA 계열 담즙산을 염증이나 부작용 없이 효율적으로 폐 조직내까지 치료적 활성량으로 전달해 폐 질환을 개선시키고 사이토카인 폭풍을 억제시킬 수 있다.As described above, in the present application, by including a water-solubilized UDCA clean aqueous solution formulation that significantly improves the pharmacokinetic properties of UDCA-based bile acids in lung tissue only by oral administration, it is possible to rapidly prevent or treat sepsis, acute lung injury, or acute respiratory distress syndrome. can That is, by oral administration of the pharmaceutical composition of the present application alone, the UDCA-based bile acid can be efficiently delivered to the lung tissue in a therapeutically active amount without inflammation or side effects, thereby improving lung disease and suppressing cytokine storm.
이에 한정되는 것은 아니나 본원의 약학 조성물은 대상에 경구투여 시 폐(lung)에서 적어도 5 분이상 동안 UDCA, tUDCA 및 gUDCA를 포함하는 UDCA 계열의 담즙산의 농도의 합이 UDCA 계열이 아닌 담즙산의 농도의 합보다 크게 될 수 있다.Although not limited thereto, when the pharmaceutical composition of the present application is orally administered to a subject, the sum of the concentrations of UDCA-based bile acids, including UDCA, tUDCA, and gUDCA, for at least 5 minutes in the lung is the same as the concentration of non-UDCA-based bile acids. can be greater than the sum.
이에 한정되는 것은 아니나, 상기 UDCA 계열의 담즙산은 UDCA이고, 약학 조성물을 대상에 경구투여시 폐(lung) 조직 내에서 1시간 이내에 UDCA 농도가 0.65± 1.03 ㎍/g tissue 이상이 되도록 할 수 있다. Although not limited thereto, the UDCA-based bile acid is UDCA, and when the pharmaceutical composition is orally administered to a subject, the UDCA concentration is 0.65± within 1 hour in the lung tissue. It can be made to be 1.03 μg/g tissue or more.
본원의 일 실시예에 의하면, 상기 UDCA 계열의 담즙산은 UDCA이고, 약학 조성물을 대상에 경구투여시 폐(lung) 조직 내에서 1시간 이내에 UDCA 농도가 7.13 ± 2.29 ㎍/g tissue를 달성될 수 있다. 즉, 본원의 약학 조성물의 UDCA는 경구투여 후 5 내지 10분이라는 단시간 내에 폐 조직 내에 분포하기 시작하여 Tmax 0.083 ± 0.0 hr에, Cmax 7.13 ± 2.29 μg/g tissue 까지 상승할 수 있으며, 일정 시간 즉 약 1시간 정도까지 머물렀다가 소실(wash-out) 될 수 있다. According to an embodiment of the present application, the UDCA-based bile acid is UDCA, and when the pharmaceutical composition is orally administered to a subject, the UDCA concentration of 7.13 ± 2.29 μg/g tissue can be achieved within 1 hour in lung tissue. . That is, the UDCA of the pharmaceutical composition of the present application starts to be distributed in the lung tissue within a short time of 5 to 10 minutes after oral administration, and can rise to Tmax 0.083 ± 0.0 hr, Cmax 7.13 ± 2.29 μg/g tissue, and for a certain period of time, that is, It can stay for up to about an hour and then wash-out.
이에 한정되는 것은 아니나, 상기 UDCA 계열의 담즙산은 tUDCA이고, 약학 조성물을 대상에 경구투여시 폐(lung) 조직 내에서 2시간 이내에 tUDCA 농도가 0.13 ± 0.17 ㎍/g tissue 이상일 수 있다.Although not limited thereto, the UDCA-based bile acid is tUDCA, and when the pharmaceutical composition is orally administered to a subject, the tUDCA concentration within 2 hours in lung tissue may be 0.13 ± 0.17 μg/g tissue or more.
상기 수가용화된 UDCA 계열 담즙산은 경구투여 시 1일 5 내지 450mg/kg의 치료학적 유효량으로 투여되도록 포함될 수 있다. 일 구현예에서, 상기 투여 형태는 UDCA로서 50 mg/kg 내지 450 mg/kg의 UDCA 또는 이의 약학적으로 허용가능한 염을 포함하는 1일 1회 이상 투여용 제제의 형태를 가지는 것이 적합할 수 있고, 50 내지 400 mg/kg/time의 UDCA 또는 이의 약학적으로 허용가능한 염을 포함하는 1일 1회 이상 투여용 제제의 형태를 가지는 것이 더 적합할 수 있다. 다른 구현예에서, 상기 투여 형태는 UDCA로서 50 내지 350 mg/Kg의 UDCA 또는 이의 약학적으로 허용가능한 염을 포함하는 1일 1회 이상 투여용 제제의 형태를 가지는 것이 더욱더 적합할 수 있다. 다른 구현예에서, 상기 투여 형태는 UDCA로서 50 내지 300 mg/Kg/time의 UDCA 또는 이의 약학적으로 허용가능한 염을 포함하는 1일 1회 이상 투여용 제제의 형태를 가지는 것이 더욱더 적합할 수 있다. 다른 구현예에서, 상기 투여 형태는 UDCA로서 50 내지 200 mg/Kg/time의 UDCA 또는 이의 약학적으로 허용가능한 염을 포함하는 1일 1회 이상 투여용 제제의 형태를 가지는 것이 더욱더 적합할 수 있다. 다른 구현예에서, 상기 투여 형태는 UDCA로서 50 내지 150 mg/Kg/time의 UDCA 또는 이의 약학적으로 허용가능한 염을 포함하는 1일 1회 이상 투여용 제제의 형태를 가지는 것이 더욱더 적합할 수 있다. 또 다른 구현예에서, 상기 투여 형태는 UDCA로서 450 mg/kg/time의 UDCA 또는 이의 약학적으로 허용가능한 염을 포함하는 1 일 1회 투여용 제제의 형태를 가질 수 있다.The water-solubilized UDCA-based bile acid may be included to be administered in a therapeutically effective amount of 5 to 450 mg/kg per day when orally administered. In one embodiment, it may be suitable for the dosage form to be in the form of a formulation for administration at least once a day including 50 mg/kg to 450 mg/kg of UDCA or a pharmaceutically acceptable salt thereof as UDCA, , It may be more suitable to have a formulation for administration at least once a day containing UDCA or a pharmaceutically acceptable salt thereof of 50 to 400 mg/kg/time. In another embodiment, it may be more suitable for the dosage form to be in the form of a formulation for administration at least once a day including 50 to 350 mg/Kg of UDCA or a pharmaceutically acceptable salt thereof as UDCA. In another embodiment, it may be more suitable for the dosage form to be in the form of a formulation for administration at least once a day including 50 to 300 mg/Kg/time of UDCA or a pharmaceutically acceptable salt thereof as UDCA. . In another embodiment, it may be more suitable for the dosage form to be in the form of a formulation for administration at least once a day containing 50 to 200 mg/Kg/time of UDCA or a pharmaceutically acceptable salt thereof as UDCA. . In another embodiment, it may be more suitable for the dosage form to be in the form of a formulation for administration at least once a day containing 50 to 150 mg/Kg/time of UDCA or a pharmaceutically acceptable salt thereof as UDCA. . In another embodiment, the dosage form may be in the form of a formulation for once-a-day administration containing 450 mg/kg/time of UDCA or a pharmaceutically acceptable salt thereof as UDCA.
이에 한정되는 것은 아니나, 경구투여 시, 바람직하게는 1일 1회 경구투여용 제제의 형태일 수 있다. Although not limited thereto, when administered orally, it may preferably be in the form of a formulation for oral administration once a day.
이에 한정되는 것은 아니나, 제1 제제의 경구투여 간격은 증상에 따라서 1일, 2일, 3일, 4일, 5일, 6일, 또는 7일 간격으로 투여할 수 있다. Although not limited thereto, the oral administration interval of the first agent may be administered at intervals of 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days depending on the symptoms.
이에 한정되는 것은 아니나, 본원의 약학 조성물은 5일 이상 그리고 1일 1회 이상 투여되는 것일 수 있다. 일 구현예에서, 상기 약학 조성물은 5일 이상 30일까지 투여되는 것이 적합할 수 있다. 다른 구현예에서, 상기 약학 조성물은 10일 이상 30일까지 투여되는 것이 더 적합할 수 있다. 다른 구현예에서, 상기 약학 조성물은 10일 이상 20일까지 투여되는 것이 더욱더 적합할 수 있다.Although not limited thereto, the pharmaceutical composition of the present application may be administered for at least 5 days and at least once a day. In one embodiment, it may be suitable for the pharmaceutical composition to be administered for 5 to 30 days. In another embodiment, the pharmaceutical composition may be more suitable to be administered for at least 10 days and up to 30 days. In another embodiment, it may be more suitable for the pharmaceutical composition to be administered for at least 10 days and up to 20 days.
본원의 약학 조성물은 정맥주사로 투여될 수 있다. 본원의 약학 조성물은 정맥주사를 통해 UDCA 담즙산이 폐로 신속하게 전달될 수 있고 혈관을 막거나 염증, 발적 등이 문제를 일으키지 않는다. 또한, 상기 약학 조성물은 폐 조직내 정맥주사용(IV injection) 제제일 수 있고, 상기 조성물의 UDCA 계열 담즙산은 폐 조직내 까지 전달될 수 있으며 주사된 후 폐 조직내에서 피부발적 또는 염증을 일으키지 않는다. 이에 한정되는 것은 아니나, 상기 수가용화된 UDCA 계열 담즙산은 정맥주사로 투여 시 1일 1.0 내지 100mg/kg의 치료학적 유효량으로 투여되도록 포함될 수 있다. 일 구현예에서, 수가용화된 UDCA 계열 담즙산은 경구투여 시, 1.0 내지 80mg의 치료학적 유효량으로 투여되는 것이 적합할 수 있다. 다른 구현예에서, 수가용화된 UDCA 계열 담즙산은 5 내지 80mg의 치료학적 유효량으로 투여되는 것이 더 적합할 수 있다. 다른 구현예에서, 수가용화된 UDCA 계열 담즙산은 10 내지 60mg의 치료학적 유효량으로 투여되는 것이 더욱 적합할 수 있다. 다른 구현예에서, 수가용화된 UDCA 계열 담즙산은 20 내지 60mg의 치료학적 유효량으로 투여되는 것이 더욱더 적합할 수 있다. 또 다른 구현예에서, 25 내지 60mg의 치료학적 유효량으로 투여되는 것이 더욱더 적합할 수 있다.The pharmaceutical composition of the present application may be administered intravenously. The pharmaceutical composition of the present application can rapidly deliver UDCA bile acid to the lungs through intravenous injection, and does not cause problems such as blockage of blood vessels, inflammation, or redness. In addition, the pharmaceutical composition may be an intravenous (IV injection) formulation in the lung tissue, and the UDCA-based bile acid of the composition may be delivered to the lung tissue and does not cause skin redness or inflammation in the lung tissue after injection. . Although not limited thereto, the water-solubilized UDCA-based bile acid may be included to be administered in a therapeutically effective amount of 1.0 to 100 mg/kg per day when administered intravenously. In one embodiment, it may be suitable to administer the water-solubilized UDCA-based bile acid in a therapeutically effective amount of 1.0 to 80 mg when orally administered. In another embodiment, it may be more suitable for the water-solubilized UDCA-based bile acid to be administered in a therapeutically effective amount of 5 to 80 mg. In another embodiment, it may be more suitable to administer the water-solubilized UDCA family bile acid in a therapeutically effective amount of 10 to 60 mg. In another embodiment, it may be more suitable for the water-solubilized UDCA family bile acid to be administered in a therapeutically effective amount of 20-60 mg. In another embodiment, it may be even more suitable to be administered in a therapeutically effective amount of 25 to 60 mg.
본원의 약학 조성물은 비강내로 투여될 수 있다. 본원의 약학 조성물은 비강내 투여를 통해 비강점막(nasal mucosa)을 통해 UDCA 담즙산이 폐로 신속하게 전달될 수 있고 염증, 발적 등이 문제를 일으키지 않는다. 이에 한정되는 것은 아니나, 상기 수가용화된 UDCA 계열 담즙산은 비강내 투여 시 1일 1.0 내지 80mg/kg의 치료학적 유효량으로 투여되도록 포함될 수 있다. 일 구현예에서, 수가용화된 UDCA 계열 담즙산은 1일 5 내지 80mg/kg의 치료학적 유효량으로 투여되도록 포함되는 것이 적합할 수 있다. 다른 구현예에서, 수가용화된 UDCA 계열 담즙산은 1일 10 내지 60mg의 치료학적 유효량으로 투여되는 것이 더 적합할 수 있다. 다른 구현예에서, 수가용화된 UDCA 계열 담즙산은 1일 20 내지 60mg의 치료학적 유효량으로 투여되는 것이 더 적합할 수 있다. 또 다른 구현예에서, 수가용화된 UDCA 계열 담즙산은 1일 25 내지 40mg의 치료학적 유효량으로 투여되는 것이 더욱더 적합할 수 있다.The pharmaceutical composition herein may be administered intranasally. In the pharmaceutical composition of the present application, UDCA bile acid can be rapidly delivered to the lungs through the nasal mucosa through intranasal administration, and inflammation, redness, etc. do not cause problems. Although not limited thereto, the water-solubilized UDCA-based bile acid may be included to be administered in a therapeutically effective amount of 1.0 to 80 mg/kg per day when administered intranasally. In one embodiment, it may be suitable to include solubilized UDCA-based bile acid to be administered in a therapeutically effective amount of 5 to 80 mg/kg per day. In another embodiment, it may be more suitable to administer the water-solubilized UDCA-based bile acid in a therapeutically effective amount of 10 to 60 mg per day. In another embodiment, it may be more suitable to administer the water-solubilized UDCA-based bile acid in a therapeutically effective amount of 20 to 60 mg per day. In another embodiment, it may be more suitable to administer the water-solubilized UDCA family bile acid in a therapeutically effective amount of 25-40 mg per day.
이에 한정되는 것은 아니나, 상기 제1 제제는, 상기 UDCA 계열의 담즙산을 포함하는 제1 물질; 말토덱스트린, 덱스트린, 액상 포도당, 옥수수 시럽 고체, 가용성 전분, 덱스트란 및 이들의 조합에서 선택되는 1종 이상의 수가용성 전분 전화물인 제2 물질; 및 물;을 포함하고, 상기 제1 물질 및 상기 제2 물질이 모두, 선택된 pH 값 범위 내의 모든 pH 값에 대해 침전되지 않고 용액상태로 남아 있는 것일 수 있다.Although not limited thereto, the first agent may include a first substance including the UDCA-based bile acid; a second substance which is an invert product of at least one water-soluble starch selected from maltodextrin, dextrin, liquid glucose, corn syrup solid, soluble starch, dextran, and combinations thereof; and water, wherein both the first material and the second material remain in a solution state without being precipitated for all pH values within a selected pH value range.
상기 수가용화된 우르소데옥시콜산(aqueous solubilized UDCA) 계열 담즙산은 UDCA가 수가용성 전분 전화물인 말토덱스트린과 함께 물(water)에 안정화되어 결과적으로 순수 UDCA 분자의 수용성 용해도가 3,000배 이상 증가될 수 있다. 수가용화된 우르소데옥시콜산은 분자 특성상 양친매성 특성을 갖는 비이온성 분자로 수용액에 단일분자 형태로 용해되어 있기 때문에 농도 차이에 의한 빠른 분산속도로 세포 간, 세포내 확산은 물론 수동적 기전에 의해 생체 내 흡수가 되므로 UDCA의 흡수율은 획기적으로 증가된다. 결론적으로 물에 60 g/L까지 고농도로 녹아있는 UDCA을 주성분으로 포함하고 있는 수가용화된 UDCA는 가장 이상적인 다기능성 약물로서, 폐 조직내 주사, 경구투여, 정맥주사로 패혈증 또는 급성 폐손상 같은 패혈증 또는 급성 폐손상 질환을 예방 또는 치료할 수 있다. In the aqueous solubilized UDCA-based bile acid, UDCA is stabilized in water together with maltodextrin, a water-soluble starch inversion product, and as a result, the aqueous solubility of pure UDCA molecules can be increased 3,000 times or more. . Water-soluble ursodeoxycholic acid is a nonionic molecule with amphiphilic properties due to its molecular nature, and is dissolved in an aqueous solution in the form of a single molecule. Because it is absorbed, the absorption rate of UDCA is dramatically increased. In conclusion, water-solubilized UDCA containing UDCA dissolved in water at a high concentration of up to 60 g/L as a main ingredient is the most ideal multifunctional drug. Or it can prevent or treat an acute lung injury disease.
상기 제1 물질 및 상기 제2 물질이 모두, 선택된 제1 물질 및 상기 제2 물질이 모두, 선택된 pH 값 범위 내의 모든 pH 값에 대해 침전되지 않고 용액상태로 남아 있기 위해서는 pH 값 범위에 따라 제1 물질에 대한 제2 물질의 최소 중량비가 달라질 수 있다. 상기 최소 중량비 조건을 만족시키지 못할 경우, 제1 물질이 침전이 발생할 수 있다. In order for both the first material and the second material, the selected first material and the second material, to remain in solution without precipitation for all pH values within the selected pH value range, the first material depends on the pH value range. The minimum weight ratio of the second material to the material may vary. If the minimum weight ratio condition is not satisfied, precipitation of the first material may occur.
이에 한정되는 것은 아니나, 상기 수가용성 전분 화합물은 말토덱스트린이고, 상기 UDCA에 대한 말토덱스트린의 최소 중량비는 1:30, 1:25, 1:20, 1:16, 1:15, 1:13, 1:12, 1:6일 수 있다. 상기 제1 제제에 사용되는 고분자량의 수가용성 전분 전화물의 양은, 선택된 우르소데옥시콜산에 원하는 농도 및 본원에 기재된 pH 범위에서 가용성을 갖는 양으로 정의될 수 있다. 상기 말토덱스트린의 최소량은 tUDCA 및 gUDCA의 경우에도 동일하게 적용될 수 있다. Although not limited thereto, the water-soluble starch compound is maltodextrin, and the minimum weight ratio of maltodextrin to UDCA is 1:30, 1:25, 1:20, 1:16, 1:15, 1:13, 1:12, 1:6. The amount of high molecular weight, water-soluble starch invert used in the first formulation may be defined as an amount having a desired concentration in the selected ursodeoxycholic acid and solubility in the pH range described herein. The minimum amount of maltodextrin may be equally applied to tUDCA and gUDCA.
이에 한정되는 것은 아니나, 약학 조성물의 pH 값은 3 ~ 10고, 상기 수가용성 전분 전화물은 말토덱스트린이고, 상기 UDCA 계열 담즙산에 대한 말토덱스트린의 최소 중량비는 1:16 ~ 1:30일 수 있다.Although not limited thereto, the pH value of the pharmaceutical composition is 3 to 10, the water-soluble starch inversion product is maltodextrin, and the minimum weight ratio of maltodextrin to the UDCA-based bile acid may be 1:16 to 1:30. .
이에 한정되는 것은 아니나, 상기 수가용성 전분 전화물은 말토덱스트린이고, 상기 UDCA 계열의 담즙산에 대한 상기 말토덱스트린의 최소 중량비는 pH 값 6.1 내지 9.6에서 1:12일 수 있다.Although not limited thereto, the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:12 at a pH value of 6.1 to 9.6.
이에 한정되는 것은 아니나, pH 값이 3 이상 6 미만에서, UDCA에 대한 말토덱스트린의 최소 중량비가 1:1 ~ 1:15인 경우 침전이 발생하여 청정 수용액이 아닐 수 있다. Although not limited thereto, when the pH value is 3 or more and less than 6, when the minimum weight ratio of maltodextrin to UDCA is 1:1 to 1:15, precipitation may occur, which may not be a clean aqueous solution.
이에 한정되는 것은 아니나, 상기 수가용성 전분 전화물은 말토덱스트린이고, 상기 UDCA 계열의 담즙산에 대한 상기 말토덱스트린의 최소 중량비는 pH 값 6 내지 9에서 1:13일 수 있다.Although not limited thereto, the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:13 at a pH value of 6 to 9.
이에 한정되는 것은 아니나, 약학 조성물의 pH 값은 6.5 ~ 8이고, 상기 수가용성 전분 전화물은 말토덱스트린이고, 상기 UDCA에 대한 말토덱스트린의 최소 중량비는 1:13 ~ 1:30일 수 있다Although not limited thereto, the pH value of the pharmaceutical composition is 6.5 to 8, the water-soluble starch inversion product is maltodextrin, and the minimum weight ratio of maltodextrin to UDCA may be 1:13 to 1:30.
이에 한정되는 것은 아니나, 상기 수가용성 전분 전화물은 말토덱스트린이고, 상기 UDCA 계열의 담즙산에 대한 상기 말토덱스트린의 최소 중량비는 pH 값 6 내지 9.5에서 1:15일 수 있다.Although not limited thereto, the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:15 at a pH value of 6 to 9.5.
이에 한정되는 것은 아니나, 상기 수가용성 전분 전화물은 말토덱스트린이고, 상기 UDCA 계열의 담즙산에 대한 상기 말토덱스트린의 최소 중량비는 pH 값 5.5 내지 9.4에서 1:20일 수 있다.Although not limited thereto, the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:20 at a pH value of 5.5 to 9.4.
이에 한정되는 것은 아니나, 상기 수가용성 전분 전화물은 말토덱스트린이고, 상기 UDCA 계열의 담즙산에 대한 상기 말토덱스트린의 최소 중량비는 pH 값 5.1 내지 9.6에서 1:25일 수 있다.Although not limited thereto, the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:25 at a pH value of 5.1 to 9.6.
이에 한정되는 것은 아니나, 상기 수가용성 전분 전화물은 말토덱스트린이고, 상기 UDCA 계열의 담즙산에 대한 상기 말토덱스트린의 최소 중량비는 pH 값 2.9 내지 9.0에서 1:30일 수 있다.Although not limited thereto, the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:30 at a pH value of 2.9 to 9.0.
이에 한정되는 것은 아니나, 상기 수가용성 전분 전화물은 말토덱스트린이고, 상기 UDCA 계열의 담즙산에 대한 상기 말토덱스트린의 최소 중량비는 pH 값 2.9 내지 10.2에서 1:30일 수 있다.Although not limited thereto, the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:30 at a pH value of 2.9 to 10.2.
이에 한정되는 것은 아니나, 상기 수가용성 전분 전화물은 말토덱스트린이고, 상기 UDCA 계열의 담즙산에 대한 상기 말토덱스트린의 최소 중량비는 pH 값 2.2 내지 10.8에서 1:29.8일 수 있다.Although not limited thereto, the water-soluble starch inversion product may be maltodextrin, and the minimum weight ratio of the maltodextrin to the UDCA-based bile acid may be 1:29.8 at a pH value of 2.2 to 10.8.
이에 한정되는 것은 아니나, 상기 UDCA 계열의 담즙산은 제1 제제의 총 중량에 대하여 0.01 내지 5 중량부로 포함될 수 있다. 이에 한정되는 것은 아니나, 제1 제제의 총 중량에 대하여 UDCA가 0.01 중량부 미만이면 패혈증, 급성 호흡곤란증후군, 또는 급성 폐손상 질환 예방 또는 치료 효과가 미미할 수 있고, 5 중량부 초과하는 경우 청정 수용액으로의 제조가 어려울 수 있다. 이에 한정되는 것은 아니나, 청정 수용액이 되지 않고 UDCA나 말토덱스트린이 석출되 침전이 발생하는 경우 폐 조직내 정맥주사제, 경구투여제 사용에 어려움이 있을 수 있다. 침전이 일어나면 UDCA가 물에 녹지 않고 결정형 UDCA로 존재할 수 있으며 이를 비강내 투여(intranasal administration) 또는 폐 조직내 주사 제제로 사용하면 결정형 UDCA로 인하여 폐염증이 일어날 가능성이 크다. 청정 수용액으로 제조하는 것은 특히 폐 조직내 주사제, 비강내 투여제제, 및 정맥주사제 제조시 피부발적 문제를 일으키는 결정형 UDCA를 전부 제거하기 위한 것이다.Although not limited thereto, the UDCA-based bile acid may be included in an amount of 0.01 to 5 parts by weight based on the total weight of the first agent. Although not limited thereto, if UDCA is less than 0.01 parts by weight based on the total weight of the first formulation, the prevention or treatment effect of sepsis, acute respiratory distress syndrome, or acute lung injury disease may be insignificant, and when it exceeds 5 parts by weight, clean aqueous solution It can be difficult to manufacture. Although not limited thereto, if UDCA or maltodextrin does not become a clean aqueous solution and precipitation occurs due to precipitation, it may be difficult to use intravenous or oral administration in lung tissue. When precipitation occurs, UDCA is not soluble in water and may exist as crystalline UDCA. If it is used for intranasal administration or intrapulmonary injection, pulmonary inflammation is highly likely to occur due to crystalline UDCA. Preparation as a clean aqueous solution is to remove all crystalline UDCA, which causes skin redness, especially in the preparation of lung tissue injections, intranasal administration preparations, and intravenous injection preparations.
이에 한정되는 것은 아니나, UDCA는 제1 제제의 총 중량에 대하여 0.01 ~ 5 중량부, 0.04 내지 5 중량부, 0.1 ~ 5 중량부, 1 ~ 5 중량부, 2 ~ 5 중량부, 3 ~ 5 중량부, 4 ~ 5 중량부, 0.01 ~ 3 중량부, 0.1 ~ 3 중량부, 1 ~ 3 중량부, 2 ~ 3 중량부, 0.01 ~ 2.5 중량부, 0.1 ~ 2.5 중량부, 1 ~ 2.5 중량부로 포함될 수 있다. 경구투여용인 경우, 조성물의 총 중량에 대하여 UDCA 함량은 0.04 ~ 2.5 중량부가 적합할 수 있고, 0.1 ~ 2.5 중량부가 적합하고, 1 ~ 2.5 중량부가 더 적합할 수 있다. 0.04 ~ 2.5 중량부로 포함되는 것이 더욱더 적합할 수 있다.Although not limited thereto, UDCA is 0.01 to 5 parts by weight, 0.04 to 5 parts by weight, 0.1 to 5 parts by weight, 1 to 5 parts by weight, 2 to 5 parts by weight, 3 to 5 parts by weight based on the total weight of the first agent. parts, 4 to 5 parts by weight, 0.01 to 3 parts by weight, 0.1 to 3 parts by weight, 1 to 3 parts by weight, 2 to 3 parts by weight, 0.01 to 2.5 parts by weight, 0.1 to 2.5 parts by weight, 1 to 2.5 parts by weight can For oral administration, 0.04 to 2.5 parts by weight of the UDCA content may be suitable, 0.1 to 2.5 parts by weight is suitable, and 1 to 2.5 parts by weight may be more suitable with respect to the total weight of the composition. It may be more suitable to be included in an amount of 0.04 to 2.5 parts by weight.
이에 한정되는 것은 아니나, 상기 수가용화 전분 전화물은 제1 제제의 총 중량에 대하여 1 ~ 70 중량부로 포함되는 것이 적합할 수 있다. 이에 한정되는 것은 아니나, 수가용화 전분 전화물은 1 중량부 미만으로 포함되면 UDCA를 유효량으로 물에 용해시킬 수 없어 패혈증, 급성 폐손상 질환 또는 급성 호흡곤란증 증후군 예방 또는 치료 효과가 미미할 수 있고, 70 중량부 초과의 경우 침전이 생겨 UDCA나 말토덱스트린이 수용액 밖으로 석출되어 이로 인한 폐에 발적이 발생할 수 있다. Although not limited thereto, it may be suitable to include the water-soluble starch inversion product in an amount of 1 to 70 parts by weight based on the total weight of the first formulation. Although not limited thereto, when the water-soluble starch inversion product is included in an amount of less than 1 part by weight, UDCA cannot be dissolved in water in an effective amount, so the prevention or treatment effect of sepsis, acute lung injury disease, or acute respiratory distress syndrome may be insignificant, 70 In the case of an excess of parts by weight, precipitation may occur and UDCA or maltodextrin may precipitate out of the aqueous solution, which may cause redness in the lungs.
이에 한정되는 것은 아니나, 상기 말토덱스트린은 조성물의 총 중량에 대하여 1 ~ 60 중량부, 5 ~ 60 중량부, 10 ~ 60 중량부, 20 ~ 60 중량부, 30 ~ 60 중량부, 40 ~ 60 중량부, 50 ~ 60중량부, 1 ~ 50 중량부, 5 ~ 50 중량부, 10 ~ 50 중량부, 20 ~ 50 중량부, 30 ~ 50 중량부, 40 ~ 50 중량부, 1 ~ 40 중량부, 5 ~ 40 중량부, 10 ~ 40 중량부, 20 ~ 40 중량부, 30 ~ 40 중량부, 1 ~ 30 중량부, 5 ~ 30 중량부, 10 ~ 30 중량부, 20 ~ 30 중량부, 1 ~ 20 중량부, 5 ~ 20 중량부, 10 ~ 20 중량부, 1 ~ 10 중량부, 5 ~ 10 중량부로 포함할 수 있다.Although not limited thereto, the maltodextrin may contain 1 to 60 parts by weight, 5 to 60 parts by weight, 10 to 60 parts by weight, 20 to 60 parts by weight, 30 to 60 parts by weight, 40 to 60 parts by weight based on the total weight of the composition. parts, 50-60 parts by weight, 1-50 parts by weight, 5-50 parts by weight, 10-50 parts by weight, 20-50 parts by weight, 30-50 parts by weight, 40-50 parts by weight, 1-40 parts by weight, 5 to 40 parts by weight, 10 to 40 parts by weight, 20 to 40 parts by weight, 30 to 40 parts by weight, 1 to 30 parts by weight, 5 to 30 parts by weight, 10 to 30 parts by weight, 20 to 30 parts by weight, 1 to 20 parts by weight, 5 to 20 parts by weight, 10 to 20 parts by weight, 1 to 10 parts by weight, 5 to 10 parts by weight may be included.
본원의 상기 수가용성 전분 전화물은 각종 pH 조건하에서 전분의 부분 또는 불완전 가수분해로부터 직접 얻어진 탄수화물을 포함한다. 비제한적인 예로는 말토덱스트린, 덱스트린, 액체 포도당, 콘 시럽 고형물(액체 포도당의 건조 분말) 및 이의 조합일 수 있다. 상기 콘 시럽 고형물은 Maltrin®　 M200이고, 말토덱스트린은 Maltrin®　M700 일 수 있고, 이에 한정하는 것은 아니나, 미국 아이오와주의 머스커틴에 소재한 Grain Processing Corporation사의 상품명 GPC로 제조된 것일 수 있다. The water-soluble starch inversion product of the present application includes carbohydrates obtained directly from partial or incomplete hydrolysis of starch under various pH conditions. Non-limiting examples may be maltodextrin, dextrin, liquid glucose, corn syrup solids (dry powder of liquid glucose), and combinations thereof. The corn syrup solid may be Maltrin ® M200, and the maltodextrin may be Maltrin ® M700, but is not limited thereto, and may be manufactured under the trade name GPC of Grain Processing Corporation, Muscatine, Iowa, USA.
본원의 상기 수가용성 전분 전화물이 중합체로 이루어진 경우, 이 중합체는 적어도 하나의 환원 단부와 적어도 하나의 비환원 단부를 가질 수 있으며, 직쇄 또는 분기쇄 일 수 있다. 또, 분자량은 약 100 질량단위 이상, 또는 106 질량단위 이상일 수 있다. 이에 한정되는 것은 아니나, 고분자량의 수가용성 전분 전화물은 분자량 105 질량단위 이상이 적합할 수 있다. 상기 조성물은 수가용성 전분 전화물 및/또는 가용성 비전분 다당류의 배합물을 포함하는 수성 용액인 조성물이 제공될 수 있다. When the water-soluble starch inversion product of the present application is made of a polymer, the polymer may have at least one reducing end and at least one non-reducing end, and may be straight-chain or branched. In addition, the molecular weight may be about 100 mass units or more, or 106 mass units or more. Although not limited thereto, the high molecular weight water-soluble starch conversion product may have a molecular weight of 105 mass units or more. The composition may be provided as an aqueous solution comprising a combination of water-soluble starch invert and/or soluble non-starch polysaccharide.
본원의 제1 제제는 가용성 비전분 다당류를 더 포함할 수 있다. 상기 수가용성 비전분 다당류는 각종 가수분해 또는 합성 기전에 의해 각종 pH 조건하에서 얻어질 수 있다. 비제한적인 예로는 덱스트란, 구아 고무, 펙틴, 난소화성의 가용성 섬유 등을 들 수 있다. The first formulation of the present application may further include a soluble non-starch polysaccharide. The water-soluble non-starch polysaccharide can be obtained under various pH conditions by various hydrolysis or synthesis mechanisms. Non-limiting examples include dextran, guar gum, pectin, indigestible soluble fiber, and the like.
본원의 제1 제제의 침전을 방지하는데 필요한 UDCA에 대한 액체 포도당(예를 들어, 콘 시럽)의 최소 중량비는 약 1:25(즉, 물 100㎖ 중 UDCA 500 ㎎ 당 약 12.5 g일 수 있고, 물 200 ㎖ 중 UDCA 1 g 당 약 25 g)일 수 있으나 이에 한정하는 것은 아니다. The minimum weight ratio of liquid glucose (e.g., corn syrup) to UDCA required to prevent precipitation of the first formulation herein is about 1:25 (i.e., about 12.5 g per 500 mg of UDCA in 100 ml of water, about 25 g per 1 g of UDCA in 200 ml of water), but is not limited thereto.
또한, 본원의 제1 제제의 투여 형태로부터, 침전을 방지하는데 필요한 액체 포도당의 건조 분말(콘 시럽 고형물, 예를 들어, Maltrin®　M200)의 최소량은 물 100 ㎖ 중 UDCA 1 g 당 약 30 g일 수 있고, 물 200 ㎖ 중 UDCA 2 g 당 약 60 g일 수 있으나, 이에 한정하는 것은 아니다.Also, from the dosage form of the first agent herein, the minimum amount of dry powder of liquid glucose (corn syrup solids, e.g., Maltrin ® M200) required to prevent precipitation is about 30 g per gram of UDCA in 100 ml of water. and may be about 60 g per 2 g of UDCA in 200 ml of water, but is not limited thereto.
또한, 본원의 제1 제제의 투여 형태로부터, 침전을 방지하는데 필요한 가용성 비전분 다당류의 최소량은 물 100 ㎖ 중 우르소데옥시콜산 500 ㎎ 당 구아 고무 약 50 g일 수 있고, 물 100 ㎖ 중 우르소데옥시콜산 500 ㎎ 당 펙틴 80 g일 수 있다. 단, 고분자량 수가용성 전분 전화물 또는 가용성 비전분 다당류의 최소 필요량은 주로 농도보다 용액 제제에서의 UDCA의 절대량에 의해 결정될 수 있다.Also, from the dosage form of the first formulation herein, the minimum amount of soluble non-starch polysaccharide required to prevent precipitation may be about 50 g of guar gum per 500 mg of ursodeoxycholic acid in 100 ml of water, and ursode in 100 ml of water. 80 g of pectin per 500 mg of oxycholic acid. However, the minimum required amount of high molecular weight water-soluble starch invert or soluble non-starch polysaccharide can be determined mainly by the absolute amount of UDCA in the solution formulation rather than the concentration.
본원의 약학 조성물은 경구투여용으로 제제화 할 경우 식이섬유를 더 포함할 수 있다. 식이섬유의 비제한적 예로는 구아고무, 펙틴, 금불초(psyllium), 귀리 고무, 콩 섬유, 귀리겨, 옥수수피, 셀룰로오스 및 소맥겨를 들 수 있으나 이에 한정하는 것은 아니다.The pharmaceutical composition of the present application may further include dietary fiber when formulated for oral administration. Non-limiting examples of dietary fiber include, but are not limited to, guar gum, pectin, psyllium, oat gum, soybean fiber, oat bran, corn hull, cellulose and wheat bran.
본원의 약학 조성물은 유화제 및 현탁제를 더 포함할 수 있다. 유화제의 비제한적인 예로는 구아 고무, 펙틴, 아카시아, 카라기난, 카복시메틸 셀룰로오스 나트륨, 하이드록시메틸 셀룰로오스, 하이드록시프로필 셀룰로오스, 메틸 셀룰로오스, 폴리비닐 알코올, 포비돈, 트래거캔스 고무, 산탄 검 및 소르비탄 에스테르를 들 수 있다.The pharmaceutical composition of the present application may further include an emulsifying agent and a suspending agent. Non-limiting examples of emulsifiers include guar gum, pectin, acacia, carrageenan, sodium carboxymethyl cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, methyl cellulose, polyvinyl alcohol, povidone, gum tragacanth, xanthan gum and sorbitan. esters.
본원의 약학 조성물은 약학적으로 허용 가능한 첨가제를 더 포함할 수 있으며, 약학적으로 허용 가능한 첨가제로는 전분, 젤라틴화 전분, 미결정셀룰로오스, 유당, 포비돈, 콜로이달실리콘디옥사이드, 인산수소칼슘, 락토스, 만니톨, 엿, 아라비아고무, 전호화전분, 옥수수전분, 분말셀룰로오스, 히드록시프로필셀룰로오스, 오파드라이, 전분글리콜산나트륨, 카르나우바 납, 합성규산알루미늄, 스테아린산, 스테아린산마그네슘, 스테아린산 알루미늄, 스테아린산칼슘, 백당, 덱스트로스, 소르비톨 및 탈크 등이 사용될 수 있다. 본원에 있어서, 약학적으로 허용 가능한 첨가제는 상기 조성물에 대해 0.1 ~ 90 중량%로 포함되는 것이 적합하나 이에 한정되는 것은 아니다. 또한, 본원의 약학 조성물은 실제 임상 투여 시에 비경구의 여러 가지 제형으로 투여될 수 있으며, 제제화 할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제될 수 있다. 비경구투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제, 주사제가 포함될 수 있다. The pharmaceutical composition of the present application may further include pharmaceutically acceptable additives, and pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, Mannitol, syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry, sodium starch glycolate, lead carnauba, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, Sucrose, dextrose, sorbitol and talc and the like can be used. In the present application, the pharmaceutically acceptable additive is preferably included in an amount of 0.1 to 90% by weight based on the composition, but is not limited thereto. In addition, the pharmaceutical composition of the present application may be administered in various parenteral formulations during actual clinical administration, and when formulated, commonly used fillers, extenders, binders, wetting agents, disintegrants, diluents such as surfactants or excipients are used. can be prepared. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and injections.
본원의 약학 조성물의 pH는 2.9 ~ 10.2일 수 있고, 상기 pH 값에서 상기 조성물이 용해된 투명한(clear) 수용액 상태일 수 있다. 상기 제1 제제는 물속에 가용화될 수 있고, 상기의 pH에서 침전 없이 청정 수용액 상태일 수 있다. 즉, 상기 제1 제제는 수개월 (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 또는 12개월) 이후에도 UDCA 계열 담즙산이 침전되지 않고 청정 수용액 상태일 수 있다. 상기 화합물을 침전시키지 않는 선택된 pH 범위는 이에 한정되는 것은 아니나, 약 pH 2.9 내지 약 pH 10.2 일 수 있으며, 약 pH 3 내지 약 pH 9가 적합하고, 이에 한정되는 것은 아니나, pH 6 내지 8이 더 적합하고, pH 6.5 내지 8이 더 적합하다. 또한, 이를 상기 pH를 유지하기 위해 필요한 경우, 산, 염기 및 완충제를 함유할 수 있다. 상기 pH 조정 물질은 이에 한정하는 것은 아니나, HCl, H3PO4, H2SO4, HNO3, CH3COOH, 구연산, 말산, 주석산, 락트산(lactic acid), 인산염, 에이데트산(eidetic acid) 및 알칼리류일 수 있다. 이러한 pH 조정 물질의 성질 및 사용 방식은 본 분야에 잘 알려져 있다. 상기 pH 범위는 투여 방법에 따라 다양한 제형이 제제로부터 용액 상태로 유지되고, 폐 조직에 주사되거나 경구투여로 혈액내로 흡수되기에 충분한 수계에서 얻어질 수 있는 임의의 하위세트(subset)의 pH 레벨이다. 따라서, 상기 조성물은 본원에 따른 조성물이 구강, 위 및 장에서 우세한 pH 레벨에서 침전되는 일없이 용액 상태에 있는 제형으로서 사용될 수 있다. The pH of the pharmaceutical composition of the present application may be in a range of 2.9 to 10.2, and may be in a clear aqueous solution state in which the composition is dissolved at the pH value. The first agent may be solubilized in water, and may be in a clean aqueous solution state without precipitation at the above pH. That is, the first formulation may be in a clean aqueous solution state without precipitation of UDCA-based bile acids even after several months (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months). The selected pH range that does not precipitate the compound may be, but is not limited to, about pH 2.9 to about pH 10.2, and suitable, but not limited to, about pH 3 to about pH 9, pH 6 to 8 is more suitable, more preferably pH 6.5 to 8. It may also contain acids, bases and buffers if necessary to maintain the above pH. The pH adjusting material is not limited thereto, but HCl, H 3 PO 4 , H 2 SO 4 , HNO 3 , CH 3 COOH, citric acid, malic acid, tartaric acid, lactic acid, phosphate, edetic acid (eidetic acid) ) and alkalis. The nature and mode of use of such pH adjusting substances are well known in the art. The above pH range is any subset of pH levels obtainable in an aqueous system that, depending on the method of administration, is sufficient for the various formulations to be maintained in solution from the formulation, to be injected into lung tissue or to be absorbed into the blood by oral administration. . Thus, the composition can be used as a formulation in which the composition according to the invention is in solution without precipitation at the pH levels prevailing in the oral cavity, stomach and intestines.
본원의 일부 실시예에 따른 UDCA 계열 담즙산은 산성 조건하에서 일반적으로 불용성임에도 불구하고 유리 상태로서 산성 조건하에 용해된 채로 있게 된다. 본원의 약학 조성물에는 제형에 첨가될 경우 가용성이 유지되는 형태의 조성물이 더 포함될 수 있다. 또한, 상기 약학 조성물은 폐 조직내 주사제, 경구투여제, 비강내 주사제 또는 정맥주사제 형태로 패혈증 또는 급성 폐손상 질환과 같은 패혈증 또는 급성 폐손상 질환의 예방 또한 치료용 조성물을 제공하기 위하여 투명하고 안정한 용액을 제공할 수 있다. The UDCA family of bile acids according to some embodiments of the present application remain dissolved under acidic conditions as free, although generally insoluble under acidic conditions. The pharmaceutical composition of the present application may further include a composition in which solubility is maintained when added to the formulation. In addition, the pharmaceutical composition is in the form of intrapulmonary injection, oral administration, intranasal injection or intravenous injection, such as sepsis or acute lung injury disease, prevention of sepsis or acute lung injury disease In order to provide a transparent and stable composition for treatment A solution may be provided.
이에 한정되는 것은 아니나, 상기 제1 제제는 액상, 시럽, 또는 건조된 제제 형태일 수 있다. 상기 시럽은 일반 시럽 또는 농후한 시럽일 수 있다.Although not limited thereto, the first formulation may be in the form of a liquid, syrup, or dried formulation. The syrup may be a regular syrup or a thick syrup.
본원의 약학 조성물은 건조하여 분말 형태로 제제화될 수 있다. 상기 분말형태의 조성물은 보관 및 취급이 용이하며, 원하는 형태의 제형의 조성물로 제조 시 용이한 이점이 있다.The pharmaceutical composition of the present application may be dried and formulated in powder form. The composition in powder form is easy to store and handle, and there is an advantage in that it is easy to prepare a composition in a desired form.
본 발명의 발명자는 선행 연구를 통해 상기와 같은 특성을 지닌 UDCA의 수가용성을 증가시켜 고농도로 용해된 형태의 수용액을 제조하는 방법을 확립하였다. 이때 UDCA 계열 담즙산은 특정 수가용성 전분 전화물 및 물을 일정비로 포함함으로써 투명한 청정 UDCA 수용액 원액(stock solution)을 제조하였다. 본원의 제1 제제인 수가용화 UDCA 계열 담즙산의 청정 수용액의 제조방법은 대한민국 특허공개 10-2018-0092886호에 개시된 방법을 참고할 수 있으며, 상기 특허문헌 전체가 본 명세서 내에 참조로 포함된다.The inventor of the present invention established a method for preparing an aqueous solution in a dissolved form at a high concentration by increasing the water solubility of UDCA having the above characteristics through prior research. At this time, a clear and clean UDCA aqueous solution stock solution was prepared by including specific water-soluble starch inversion water and water in a certain ratio for UDCA-based bile acids. For a method for preparing a clean aqueous solution of water-solubilized UDCA-based bile acids, which is the first formulation of the present application, reference may be made to the method disclosed in Korean Patent Publication No. 10-2018-0092886, the entire patent document is incorporated herein by reference.
본원의 약학 조성물은 다른 패혈증, 급성 폐손상 질환, 또는 급성 호흡곤란증증후군 치료제와 병용투여될 수 있다.The pharmaceutical composition of the present application may be administered in combination with other sepsis, acute lung injury disease, or a therapeutic agent for acute respiratory distress syndrome.
이에 한정되는 것은 아니나, 상기 패혈증, 급성 폐손상 질환, 또는 급성 호흡곤란증후군 치료제는 정맥 주사용일 수 있다.Although not limited thereto, the therapeutic agent for sepsis, acute lung injury, or acute respiratory distress syndrome may be for intravenous injection.
본원은 패혈증, 급성 폐손상 질환, 또는 급성 호흡곤란증후군의 유전학, 생화학 및 세포 생물학의 현행의 인식에 대한 광범위한 정보를 포함하지만, 앞으로의 연구는 이들 인식의 측면들이 부정확하거나 불완전한 것임을 드러낼 수도 있다. 따라서, 통상의 기술자라면 본 발명의 일부가 취해지든 전부가 취해지든지 간에 특정 모델이나 작용 기전으로 제한되지 않은 것임을 명기한다. Although this application contains extensive information on current perceptions of the genetics, biochemistry and cellular biology of sepsis, acute lung injury disease, or acute respiratory distress syndrome, future research may reveal that aspects of these perceptions are inaccurate or incomplete. . Accordingly, it is stated to those skilled in the art that the present invention is not limited to any particular model or mechanism of action, whether in part or in whole.
본원의 다른 측면으로, 기타 등가의 또는 대안적인 조성물 및 방법이 이용될 수 있다. 예를 들어, 다수의 화합물이 수가용화된 우르소데옥시콜산과 함께 투여될 수 있는 것으로 설명하였으나, 기타 다른 화합물도 포함될 수 있다. In other aspects of the present disclosure, other equivalent or alternative compositions and methods may be used. For example, although it has been described that many of the compounds may be administered with solubilized ursodeoxycholic acid, other compounds may be included.
본 발명의 이해를 돕기 위하여 실시예를 제시한다. 하기 실시예는 본 발명을 보다 쉽게 이해하기 위하여 제공되는 것일 뿐, 실시예에 의해 본 발명의 범위가 한정되는 것은 아니다. Examples are provided to help the understanding of the present invention. The following examples are provided only for easier understanding of the present invention, and the scope of the present invention is not limited by the examples.
<실시예 1> 수가용화된 UDCA 청정 수용액 제조><Example 1> Preparation of water-solubilized UDCA clean aqueous solution>
1-1. UDCA에 대한 말토텍스트린의 함량비가 1:6 인 수가용화된 UDCA 청정 수용액1-1. Solubilized UDCA clean aqueous solution with a content ratio of maltotextrin to UDCA of 1:6
수산화나트륨 펠릿 6.7g을 정제수 400 ㎖에 용해시켰다. 다음에, UDCA 60g을 실온에서 교반하에 상기 수산화나트륨 용액에 용해시켰다. 이어서, 상기 투명한 용액에 말토덱스트린 360g을 조금씩 첨가하고 교반하였다. 이어서, 높은 처리량으로 초음파 분해(750W, 2OkHz)를 실시하면서 얻어진 투명한 용액에 방부제를 약제학적 제형에 적합한 양으로 첨가하고 HCl의 적가에 의해 pH를 조정하였다. 정제수를 첨가하여 총 1,000 ㎖까지 조정하였다. 필요한 경우, 상기 투명한 용액을 적절한 여과장치에 의해서 여과하였다. 이 여과는 원료에서 따라오는 불순물 제거나 멸균을 위해서 중요하지만, 용액이 이미 투명하므로 입상물을 제거하기 위한 것은 아니다. 표 1에서와 같이, 상기 제조된 우르소데옥시콜산 용액은 pH 10.3, 9.2, 6.7에서 육안상 침전 없이 청정 수용액을 형성하였으나 pH 5.4에서는 침전을 형성하였다.수산화나트륨 펠릿 6.7g을 정제수 400 ㎖에 용해시켰다. 다음에, UDCA 60g을 실온에서 교반하에 상기 수산화나트륨 용액에 용해시켰다. 이어서, 상기 투명한 용액에 말토덱스트린 360g을 조금씩 첨가하고 교반하였다. 이어서, 높은 처리량으로 초음파 분해(750W, 2OkHz)를 실시하면서 얻어진 투명한 용액에 방부제를 약제학적 제형에 적합한 양으로 첨가하고 HCl의 적가에 의해 pH를 조정하였다. 정제수를 첨가하여 총 1,000 ㎖까지 조정하였다. 필요한 경우, 상기 투명한 용액을 적절한 여과장치에 의해서 여과하였다. 이 여과는 원료에서 따라오는 불순물 제거나 멸균을 위해서 중요하지만, 용액이 이미 투명하므로 입상물을 제거하기 위한 것은 아니다. 표 1에서와 같이 상기 제조된 우르소데옥시콜산 용액은 pH 10.3, 9.2, 6.7에서 육안상 침전 없이 청정 수용액을 형성하였으나 pH 5.4에서는 침전을 형성하였다.6.7 g of sodium hydroxide pellets were dissolved in 400 ml of purified water. Then, 60 g of UDCA was dissolved in the sodium hydroxide solution at room temperature under stirring. Then, 360 g of maltodextrin was added little by little to the clear solution and stirred. Then, a preservative was added in an amount suitable for a pharmaceutical formulation to the clear solution obtained while performing sonication at a high throughput (750W, 20kHz), and the pH was adjusted by dropwise addition of HCl. Purified water was added to adjust the total to 1,000 ml. If necessary, the clear solution was filtered through an appropriate filter device. This filtration is important for sterilization or removal of impurities from the raw material, but not for removing particulate matter since the solution is already transparent. As shown in Table 1, the prepared ursodeoxycholic acid solution formed a clear aqueous solution without visual precipitation at pH 10.3, 9.2, and 6.7, but formed a precipitate at pH 5.4. Dissolve 6.7 g of sodium hydroxide pellets in 400 ml of purified water made it Then, 60 g of UDCA was dissolved in the sodium hydroxide solution at room temperature under stirring. Then, 360 g of maltodextrin was added little by little to the clear solution and stirred. Then, a preservative was added in an amount suitable for a pharmaceutical formulation to the clear solution obtained while performing sonication at a high throughput (750W, 20kHz), and the pH was adjusted by dropwise addition of HCl. Purified water was added to adjust the total to 1,000 ml. If necessary, the clear solution was filtered through an appropriate filter device. This filtration is important for sterilization or removal of impurities from the raw material, but not for removing particulate matter since the solution is already transparent. As shown in Table 1, the prepared ursodeoxycholic acid solution formed a clear aqueous solution without visual precipitation at pH 10.3, 9.2, and 6.7, but formed a precipitate at pH 5.4.
Figure PCTKR2021012692-appb-T000001
Figure PCTKR2021012692-appb-T000001
1-2. UDCA에 대한 말토텍스트린의 함량비가 1:12 인 수가용화된 UDCA 청정 수용액1-2. Solubilized UDCA clean aqueous solution with a content ratio of maltodextrin to UDCA of 1:12
자연 그대로의 UDCA 및 낮은 포도당 당량을 지닌 수가용성 전분을 포함하는 수가용화된 UDCA의 투명한 청정 수용액 원액(stock solution)을 제조하였다.A clear, clean aqueous stock solution of water-solubilized UDCA containing raw UDCA and water-soluble starch with a low glucose equivalent was prepared.
구체적으로 설명하면, UDCA 60g당 하나의 고분자량 수가용성 전분 전화물로서의 말토덱스트린 720g을 사용하였다는 것을 제외하고는 실시예 1-1과 동일한 가이드라인에 따라 제조되었다. 표 1에서와 같이 상기 제조된 UDCA 용액은 pH 9.6, 7.3, 6.5, 6.1에서 육안상 침전 없이 청정 수용액을 형성하였으나 pH 5.5에서는 침전을 형성하였다.Specifically, it was prepared according to the same guidelines as in Example 1-1, except that 720 g of maltodextrin as one high molecular weight water-soluble starch inversion product per 60 g of UDCA was used. As shown in Table 1, the prepared UDCA solution formed a clear aqueous solution without visual precipitation at pH 9.6, 7.3, 6.5, and 6.1, but formed a precipitate at pH 5.5.
1-3. UDCA에 대한 말토텍스트린의 함량비가 1:15 인 수가용화된 UDCA 청정 수용액1-3. Solubilized UDCA clean aqueous solution with a content ratio of maltotextrin to UDCA of 1:15
자연 그대로의 UDCA 및 낮은 포도당 당량을 지닌 수가용성 전분을 포함하는 수가용화된 UDCA의 투명한 청정 수용액 원액(stock solution)을 제조하였다.A clear, clean aqueous stock solution of water-solubilized UDCA containing raw UDCA and water-soluble starch with a low glucose equivalent was prepared.
구체적으로 설명하면, UDCA 50g당 하나의 고분자량 수가용성 전분 전화물로서의 말토덱스트린 750g을 사용하였다는 것을 제외하고는 실시예 1-1과 동일한 가이드라인에 따라 제조되었다. 이때 수산화나트륨 펠릿 5.7g을 정제수 400 ㎖에 용해시킨 다음 사용하였다. 표 1에서와 같이 상기 제조된 우르소데옥시콜산 용액은 pH 9.5, 8.9, 7.9, 7.1, 6.0에서 육안상 침전 없이 청정 수용액을 형성하였다. 그러나 pH 5.5에서는 침전을 형성하였다. 도 1은 각 pH값에서의 UDCA 용액을 테스트 튜브에 담아 청정 수용액 생성여부를 사진으로 나타낸 것이다.Specifically, it was prepared according to the same guidelines as in Example 1-1, except that 750 g of maltodextrin as one high molecular weight water-soluble starch invert per 50 g of UDCA was used. At this time, 5.7 g of sodium hydroxide pellets were dissolved in 400 ml of purified water and then used. As shown in Table 1, the prepared ursodeoxycholic acid solution formed a clear aqueous solution without visual precipitation at pH 9.5, 8.9, 7.9, 7.1, and 6.0. However, a precipitate was formed at pH 5.5. 1 is a photograph showing whether a clean aqueous solution is generated by putting a UDCA solution at each pH value in a test tube.
1-4. UDCA에 대한 말토텍스트린의 함량비가 1:20 인 수가용화된 UDCA 청정 수용액1-4. Solubilized UDCA clean aqueous solution with a content ratio of maltotextrin to UDCA of 1:20
자연 그대로의 UDCA 및 낮은 포도당 당량을 지닌 수가용성 전분을 포함하는 수가용화된 UDCA의 투명한 청정 수용액 원액(stock solution)을 제조하였다.A clear, clean aqueous stock solution of water-solubilized UDCA containing raw UDCA and water-soluble starch with a low glucose equivalent was prepared.
구체적으로 설명하면, UDCA 17.5g당 하나의 고분자량 수가용성 전분 전화물로서의 말토덱스트린 350g을 사용하였다는 것을 제외하고는 실시예 1-1과 동일한 가이드라인에 따라 제조되었다. 이때 수산화나트륨 펠릿 2.0g을 정제수 400 ㎖에 용해시킨 다음 사용하였다. 표 1에서와 같이 상기 제조된 UDCA 용액은 pH 9.4, 7.1, 6.1, 5.5에서 육안상 침전 없이 청정 수용액을 형성하였다. 그러나 pH 5.1에서는 침전을 형성하였다. 도 2는 각 pH값에서의 UDCA 용액을 테스트 튜브에 담아 청정 수용액 생성여부를 사진으로 나타낸 것이다.Specifically, it was prepared according to the same guidelines as in Example 1-1, except that 350 g of maltodextrin as one high molecular weight water-soluble starch invert per 17.5 g of UDCA was used. At this time, 2.0 g of sodium hydroxide pellets were dissolved in 400 ml of purified water and then used. As shown in Table 1, the prepared UDCA solution formed a clear aqueous solution without visual precipitation at pH 9.4, 7.1, 6.1, and 5.5. However, a precipitate was formed at pH 5.1. 2 is a photograph showing whether a clean aqueous solution is generated by putting the UDCA solution at each pH value in a test tube.
1-5. UDCA에 대한 말토텍스트린의 함량비가 1:25 인 수가용화된 UDCA 청정 수용액1-5. A clean aqueous solution of solubilized UDCA with a content ratio of maltotextrin to UDCA of 1:25
자연 그대로의 UDCA 및 낮은 포도당 당량을 지닌 수가용성 전분을 포함하는 수가용화된 UDCA의 투명한 청정 수용액 원액(stock solution)을 제조하였다.A clear, clean aqueous stock solution of water-solubilized UDCA containing raw UDCA and water-soluble starch with a low glucose equivalent was prepared.
구체적으로 설명하면, UDCA 14g당 하나의 고분자량 수가용성 전분 전화물로서의 말토덱스트린 350g을 사용하였다는 것을 제외하고는 실시예 1-1과 동일한 가이드라인에 따라 제조되었다. 이때 수산화나트륨 펠릿 1.7g을 정제수 400 ㎖에 용해시킨 다음 사용하였다. 표 1에서와 같이 상기 제조된 UDCA 용액은 pH 9.6, 6.1, 5.1에서 육안상 침전 없이 청정 수용액을 형성하였다. 그러나 pH 4.0에서는 침전을 형성하였다. 도 3은 각 pH값에서의 UDCA 용액을 테스트 튜브에 담아 청정 수용액 생성여부를 사진으로 나타낸 것이다.Specifically, it was prepared according to the same guidelines as in Example 1-1, except that 350 g of maltodextrin as one high molecular weight water-soluble starch invert per 14 g of UDCA was used. At this time, 1.7 g of sodium hydroxide pellets were dissolved in 400 ml of purified water and then used. As shown in Table 1, the prepared UDCA solution formed a clear aqueous solution without visual precipitation at pH 9.6, 6.1, and 5.1. However, a precipitate was formed at pH 4.0. 3 is a photograph showing whether a clean aqueous solution is generated by putting the UDCA solution at each pH value in a test tube.
1-6. UDCA에 대한 말토텍스트린의 함량비가 1:30 인 수가용화된 UDCA 청정 수용액1-6. Solubilized UDCA clean aqueous solution with a content ratio of maltotextrin to UDCA of 1:30
자연 그대로의 UDCA 및 낮은 포도당 당량을 지닌 수가용성 전분을 포함하는 수가용화된 UDCA의 투명한 청정 수용액 원액(stock solution)을 제조하였다.A clear, clean aqueous stock solution of water-solubilized UDCA containing raw UDCA and water-soluble starch with a low glucose equivalent was prepared.
구체적으로 설명하면, UDCA 25g당 하나의 고분자량 수가용성 전분 전화물로서의 말토덱스트린 750g을 사용하였다는 것을 제외하고는 실시예 1-1과 동일한 가이드라인에 따라 제조되었다. 이때 수산화나트륨 펠릿 2.8g을 정제수 400 ㎖에 용해시킨 다음 사용하였다. 표 1에서와 같이 상기 제조된 UDCA 용액은 pH 9.0, 8.0, 7.0, 6.0, 5.1, 4.1, 2.9에서 육안상 침전 없이 청정 수용액을 형성하였다. 도 4는 각 pH값에서의 UDCA 용액을 테스트 튜브에 담아 청정 수용액 생성여부를 사진으로 나타낸 것이다.Specifically, it was prepared according to the same guidelines as in Example 1-1, except that 750 g of maltodextrin as one high molecular weight water-soluble starch invert per 25 g of UDCA was used. At this time, 2.8 g of sodium hydroxide pellets were dissolved in 400 ml of purified water and then used. As shown in Table 1, the prepared UDCA solution formed a clear aqueous solution without visual precipitation at pH 9.0, 8.0, 7.0, 6.0, 5.1, 4.1, and 2.9. 4 is a photograph showing whether a clean aqueous solution is generated by putting the UDCA solution at each pH value in a test tube.
1-7. UDCA/tUDCA/gUDCA를 함유하고 말토텍스트린에 대한 함량비가 1:30 인 수가용화된 UDCA 청정 수용액1-7. A clean aqueous solution of UDCA containing UDCA/tUDCA/gUDCA and having a content ratio of 1:30 to maltotextrin
UDCA과 UDCA 유도체를 함유하고 낮은 포도당 당량을 지닌 수가용성 전분을 포함하는 수가용화된 UDCA의 투명한 청정 수용액 원액(stock solution)을 제조하였다.A clear, clear aqueous stock solution of water-solubilized UDCA containing UDCA and UDCA derivatives and containing water-soluble starch with a low glucose equivalent was prepared.
구체적으로 설명하면, 수산화나트륨 펠릿 0.3g을 정제수 500 ㎖에 용해시켰다. 다음에, UDCA 1.0g, tUDCA 0.5g, gUDCA 0,5g을 실온에서 교반하에 상기 수산화나트륨 용액에 용해시켰다. 이어서, 상기 투명한 용액에 말토덱스트린 60g을 조금씩 첨가하고 교반하였다. 이어서, 높은 처리량으로 초음파 분해(750W, 2OkHz)를 실시하면서 얻어진 투명한 용액에 방부제를 약제학적 제형에 적합한 양으로 첨가하고 HCl의 적가에 의해 pH를 조정하였다. 정제수를 첨가하여 총 1,000 ㎖까지 조정하였다. 표 1에서와 같이 상기 제조된 UDCA 용액은 pH 10.2, 9.0, 8.1, 7.1, 6.1, 5.1, 4.1, 2.9에서 육안상 침전 없이 청정 수용액을 형성하였다. 도 5는 각 pH 값에서의 상기 UDCA 용액을 테스트 튜브에 담아 청정 수용액 생성여부를 사진으로 나타낸 것이다.Specifically, 0.3 g of sodium hydroxide pellets were dissolved in 500 ml of purified water. Next, 1.0 g of UDCA, 0.5 g of tUDCA, and 0.5 g of gUDCA were dissolved in the sodium hydroxide solution at room temperature under stirring. Then, 60 g of maltodextrin was added little by little to the clear solution and stirred. Then, a preservative was added in an amount suitable for a pharmaceutical formulation to the clear solution obtained while performing sonication at a high throughput (750W, 20kHz), and the pH was adjusted by dropwise addition of HCl. Purified water was added to adjust the total to 1,000 ml. As shown in Table 1, the prepared UDCA solution formed a clear aqueous solution without visual precipitation at pH 10.2, 9.0, 8.1, 7.1, 6.1, 5.1, 4.1, 2.9. 5 is a photograph showing whether a clean aqueous solution is generated by putting the UDCA solution at each pH value in a test tube.
1-8. 타우로우르소데옥시콜산(tauroursodeoxycholic acid, tUDCA)을 함유하고 말토텍스트린에 대한 함량비가 1:29.8 인 수가용화된 tUDCA 청정 수용액1-8. A clean aqueous solution of tUDCA containing tauroursodeoxycholic acid (tUDCA) and having a content ratio of 1:29.8 to maltotextrin
타우로우르소데옥시콜산(tUDCA)를 함유하고 낮은 포도당 당량을 지닌 수가용성 전분을 포함하는 수가용화된 tUDCA의 투명한 청정 수용액 원액(stock solution)을 제조하였다.A clear, clean aqueous stock solution of water-solubilized tUDCA containing tauroursodeoxycholic acid (tUDCA) and water-soluble starch with a low glucose equivalent was prepared.
구체적으로 설명하면, 수산화나트륨 펠릿 4.2g을 정제수 500 ㎖에 용해시켰다. 다음에, tUDCA 25g을 실온에서 교반하에 상기 수산화나트륨 용액에 용해시켰다. 이어서, 상기 투명한 용액에 말토덱스트린 745g을 조금씩 첨가하고 교반하였다. 이어서, 높은 처리량으로 초음파 분해(750W, 2OkHz)를 실시하면서 얻어진 투명한 용액에 방부제를 약제학적 제형에 적합한 양으로 첨가하고 HCl의 적가에 의해 pH를 조정하였다. 정제수를 첨가하여 총 1,000 ㎖까지 조정하였다. 표 1에서와 같이 상기 제조된 UDCA 용액은 pH 10.8, 9.9, 8.6, 7.5, 6.5, 5.6, 4.7, 3.5, 2.2에서 육안상 침전 없이 청정 수용액을 형성하였다. 도 6은 각 pH 값에서의 상기 tUDCA 용액을 테스트 튜브에 담아 청정 수용액 생성여부를 사진으로 나타낸 것이다.Specifically, 4.2 g of sodium hydroxide pellets were dissolved in 500 ml of purified water. Then, 25 g of tUDCA was dissolved in the sodium hydroxide solution at room temperature under stirring. Then, 745 g of maltodextrin was added little by little to the clear solution and stirred. Then, a preservative was added in an amount suitable for a pharmaceutical formulation to the clear solution obtained while performing sonication at a high throughput (750W, 20kHz), and the pH was adjusted by dropwise addition of HCl. Purified water was added to adjust the total to 1,000 ml. As shown in Table 1, the prepared UDCA solution formed a clear aqueous solution without visual precipitation at pH 10.8, 9.9, 8.6, 7.5, 6.5, 5.6, 4.7, 3.5, and 2.2. 6 is a photograph showing whether a clean aqueous solution is generated by putting the tUDCA solution at each pH value in a test tube.
<실시예 2> 패혈증, 급성 폐손상 질환, 또는 급성 호흡곤란증후군 예방 또는 치료용 수가용화된 UDCA 청정 수용액 제조<Example 2> Preparation of water-solubilized UDCA clean aqueous solution for prevention or treatment of sepsis, acute lung injury, or acute respiratory distress syndrome
2-1. 수가용화된 UDCA 청정 수용액 (시험물질 및/또는 코드명 AGP600이라고 함)2-1. Solubilized UDCA clean aqueous solution (referred to as test substance and/or codename AGP600)
상기 수가용화 UDCA(ursodeoxycholic acid) 제제는 자연 그대로의 UDCA 및 낮은 포도당 당량을 지닌 수가용성 전분을 포함하는 투명한 청정 수용액으로 먼저 수가용화 UDCA 원액(stock solution)을 제조하였다. The water-solubilized ursodeoxycholic acid (UDCA) formulation is a clear, clean aqueous solution containing pristine UDCA and water-soluble starch having a low glucose equivalent. First, a water-solubilized UDCA stock solution was prepared.
구체적으로 설명하면, UDCA 25g당 하나의 고분자량 수가용성 전분 전화물로서의 말토덱스트린(maltodextrin) 750g을 사용하였다. 수산화나트륨 펠릿 2.8g을 정제수 100 ㎖에 용해시킨 다음 UDCA 25g을 투하하고 교반해 투명한 UDCA 용액을 제조하였다(용액 1). 다른 비이커에 정제수 400ml을 준비하고 말토텍스트린 750g을 투하하고 교반해 투명한 말토덱스트린 용액을 얻었다(용액 2). 용액 1과 용액 2를 혼합해 1시간 이상 교반하면서, 필요에 따라 방부제를 약제학적 제형에 적합한 양으로 첨가하였으며, 염산을 사용하여 적가에 의해 pH를 조절하였다 (pH 6.0~7.5) 이후 의약용수를 첨가하여 총 1,000ml까지 조정하였다. 이후, 상기 투명한 용액을 무균 조건하에서 0.2㎛ 필터웨어 여과장치를 이용하여 여과 멸균 처리하거나 또는 80℃ 내지 100℃에서 가열 멸균하였다. 상기 여과는 멸균을 위해서 중요하지만, 용액이 이미 투명하므로 입상물을 제거하기 위한 것은 아니다. 상기 제조된 수가용화 UDCA 용액은 pH 12, 10, 9.0, 8.0, 7.0, 6.0, 5.0, 4.0, 3.0, 2.0에서 육안상 침전 없이 청정 수용액을 형성하였다.Specifically, 750 g of maltodextrin as one high molecular weight water-soluble starch inversion product per 25 g of UDCA was used. After dissolving 2.8 g of sodium hydroxide pellets in 100 ml of purified water, 25 g of UDCA was added thereto and stirred to prepare a transparent UDCA solution (solution 1). In another beaker, 400 ml of purified water was prepared, and 750 g of maltodextrin was added thereto, followed by stirring to obtain a transparent maltodextrin solution (solution 2). While solution 1 and solution 2 were mixed and stirred for at least 1 hour, a preservative was added in an amount suitable for the pharmaceutical formulation as needed, and the pH was adjusted by dropwise addition using hydrochloric acid (pH 6.0 to 7.5), and then the medicinal water was added and adjusted to a total of 1,000 ml. Thereafter, the transparent solution was sterilized by filtration using a 0.2 μm filterware filtration device under aseptic conditions or heat sterilized at 80° C. to 100° C. The filtration is important for sterilization, but not to remove particulates as the solution is already clear. The prepared aqueous solubilized UDCA solution formed a clear aqueous solution without visual precipitation at pH 12, 10, 9.0, 8.0, 7.0, 6.0, 5.0, 4.0, 3.0, 2.0.
<실시예 3> 내독소 유발 패혈증 동물모델에서 사이토카인 억제 및 생존율, 폐질환 개선 시험<Example 3> Cytokine inhibition and survival rate, lung disease improvement test in endotoxin-induced sepsis animal model
3-1. 패혈증 또는 급성 폐손상 동물모델 제작 및 위탁시험처 3-1. Production of sepsis or acute lung injury animal model and consignment laboratory
동물실험을 위하여는 GLP 독성평가기관인 ㈜켐온의 비임상연구소에서 실시되었으며, SOP를 준수하였다. 동물 시험계 및 사육환경으로 특정병원균 부재 (SPF) C57BL/6N 마우스, C57BL/6NCrljOri를 사용하였으며, 생산자 및 공급원은 ㈜오리엔트 바이오 (경기도 성남시 중원구 갈마치로 322)이다. 검역 및 순화를 실시하였고, ㈜켐온의 실험동물운영위원회에 의해 승인되었다. (심의번호: 2020-06-008). For animal testing, it was conducted at the non-clinical laboratory of Chemon Co., Ltd., a GLP toxicity evaluation institution, and the SOP was complied with. Specific pathogen-free (SPF) C57BL/6N mice and C57BL/6NCrljOri were used as animal test systems and breeding environments. Quarantine and acclimatization were carried out, and it was approved by the Experimental Animal Steering Committee of Chemon Co., Ltd. (Review number: 2020-06-008).
사육환경은 동물은 온도 23 ± 3 ℃, 상대습도 55 ± 15 %, 환기횟수 10-20 회/hr, 조명시간 12 시간 (오전 8 시 점등-오후 8 시 소등) 및 조도 150-300 Lux로 유지되는 경기바이오센터 동물사육구역 1 호실에서 사육하였다. The breeding environment is maintained at 23 ± 3 ℃, relative humidity of 55 ± 15 %, ventilation frequency of 10-20 times/hr, lighting time of 12 hours (lights at 8 am - lights out at 8 pm), and illuminance of 150-300 Lux. They were bred in room 1 of the animal breeding area of the Gyeonggi Bio Center.
사료는 Teklad certified irradiated global 18 % protein rodent diet (2918C, ENVIGO, UK)를 ㈜두열바이오텍 (서울특별시 서초구 바우뫼로 91 성보프라자 107 호)으로부터 공급받아 자유섭취 하도록 하였다. For the feed, Teklad certified irradiated global 18 % protein rodent diet (2918C, ENVIGO, UK) was supplied from Dooyeol Biotech Co., Ltd. (No. 107 Seongbo Plaza, 91, Baumeo-ro, Seocho-gu, Seoul) and allowed to be freely consumed.
물은 자외선 살균기 및 미세여과장치로 소독한 상수도수를 폴리카보네이트제 음수병에 넣고 자유섭취 하도록 하였다. 수질검사는 경기도보건환경연구원 (경기도 수원시 장안구 파장천로 95)에서 수행하였고, 먹는물 수질기준에 적합하였다. As for water, tap water sterilized with an ultraviolet sterilizer and microfiltration device was placed in a polycarbonate drinking water bottle and allowed to be consumed freely. The water quality test was performed at the Gyeonggi Health and Environment Research Institute (95, Pawangcheon-ro, Jangan-gu, Suwon-si, Gyeonggi-do) and was suitable for drinking water quality standards.
깔개는 고압증기멸균 후 사용하였다. 오염물질 검사는 ㈜켐온의 SOP에 따라 수행하였다. 사육상자 및 사육밀도는 폴리카보네이트 사육상자 (W 170 x L 235 x H 125 mm)에서 순화, 투여 및 관찰기간에 1 마리/사육상자로 수용하였다. 사육상자, 깔개 및 물병은 주 1 회 이상 교환하였다.The rug was used after autoclaving. Contaminant inspection was performed according to Chemon's SOP. Breeding boxes and breeding densities were housed in a polycarbonate breeding box (W 170 x L 235 x H 125 mm) at 1 animal / breeding box during the period of acclimatization, administration and observation. Breeding boxes, rugs and water bottles were exchanged at least once a week.
3-2. 내독소 유발 패혈증 동물모델에서 사이토카인 억제 효과 평가3-2. Evaluation of cytokine inhibitory effect in endotoxin-induced sepsis animal model
UDCA 및 수가용화 UDCA를 사용하여 패혈증 또는 급성 폐손상 치료효과에 대한 유효성을 확립하기 위하여 내독소 유발 패혈증 동물모델에서 시험물질 투여에 따른 사이토카인 억제 효과를 확인하였다. 즉, 수가용화 UDCA가 빠른 과도한 염증반응을 완화시켜 사이토카인 폭풍 관련 치사율을 낮추는 가능성과 작용 기전의 근거를 확보하였다. 만약 UDCA가 주요 염증유발 사이토카인의 농도를 억제할 수 있으면, 패혈증이나 급성호흡곤란성질환(ARDS) 및 신종 코로나바이러스(Covid-19) 질환에서도 UDCA 및 수가용화 UDCA 제제를 예방 및 치료제로써 사용할 수 있다. For the treatment effect of sepsis or acute lung injury using UDCA and solubilized UDCA To establish the effectiveness, the cytokine inhibitory effect of the test substance administration was confirmed in an animal model of endotoxin-induced sepsis. In other words, the possibility of water-soluble UDCA reducing the cytokine storm-related lethality by alleviating the rapid excessive inflammatory response and the basis of the mechanism of action were secured. If UDCA can inhibit the concentration of major pro-inflammatory cytokines, UDCA and water-soluble UDCA preparations can be used as prophylactic and therapeutic agents in sepsis, acute respiratory distress disease (ARDS) and novel coronavirus (Covid-19) diseases. .
LPS의 복강 투여로 유발된 패혈증 마우스 모델에서 시험물질의 6 일 반복 투여에 의한 항염증 효과를 먼저 평가하였다. 양성대조물질로 항염증 효과가 잘 알려진 스테로이드제제중 하나인 덱사메타손 (dexamethasone)을 사용하였다.In a mouse model of sepsis induced by intraperitoneal administration of LPS, the anti-inflammatory effect of 6-day repeated administration of the test substance was first evaluated. As a positive control, dexamethasone, one of the steroids with well-known anti-inflammatory effects, was used.
시험군 구성은 아래 표 2와 같다.The composition of the test group is shown in Table 2 below.
Figure PCTKR2021012692-appb-T000002
Figure PCTKR2021012692-appb-T000002
상기 시험군 G2의 부형제 대조군(수가용화 UDCA-vehicle)은 상기 실시예 2의 수가용화된 UDCA 청정 수용액에서 UDCA만을 제외하고 제조한 용액을 수가용화 UDCA-vehicle이라고 한다. 즉 비이커에 정제수 500ml을 준비하고 말토텍스트린 750g을 투하하고 교반해 투명한 말토덱스트린 용액을 얻었다. 이 용액에 방부제를 약제학적 제형에 적합한 양으로 첨가할 수 있으며, 염산을 사용하여 적가에 의해 pH를 3.0 내지 9.0 사이로 조정하였다. 이후 정제수를 첨가하여 총 1,000ml 까지 조정하였다. 필요한 경우 상기 투명한 용액을 적절한 여과장치에 의해서 여과하였다. 본 실험 G4-G6에 사용된 수가용화 UDCA-vehicle은 UDCA가 0.0mg/ml 이고, pH는 6.0~7.0 부근이었으며, 방부제는 포함하지 않았다.As for the excipient control group (water-solubilized UDCA-vehicle) of the test group G2, the solution prepared except for UDCA in the water-solubilized UDCA clean aqueous solution of Example 2 is called a water-solubilized UDCA-vehicle. That is, 500 ml of purified water was prepared in a beaker, and 750 g of maltodextrin was added thereto, followed by stirring to obtain a transparent maltodextrin solution. A preservative may be added to this solution in an amount suitable for pharmaceutical formulations, and the pH is adjusted to between 3.0 and 9.0 by dropwise addition with hydrochloric acid. Thereafter, purified water was added to adjust the total volume to 1,000 ml. If necessary, the clear solution was filtered through an appropriate filter device. The water-solubilized UDCA-vehicle used in this experiment G4-G6 had a UDCA of 0.0mg/ml, a pH of around 6.0-7.0, and no preservatives.
상기 실험에서는 수가용화된 UDCA 청정 수용액 원액 (UDCA로 25mg/ml)을 G4-G6 군에 맞게 희석하거나 투여 volume을 조절하여 실험대상 마우스에 투여하였다. G4-G6에 사용한 수가용화 UDCA(시험물질)는 25mg/ml 이고, pH는 6.0~7.0 이었으며, 방부제는 포함하지 않았다.In the above experiment, water-solubilized UDCA clean aqueous solution stock solution (25 mg/ml as UDCA) was diluted to suit the G4-G6 group or administered to the test subject mice by adjusting the administration volume. The water-solubilized UDCA (test substance) used in G4-G6 was 25 mg/ml, pH 6.0-7.0, and no preservatives were included.
투여방법은 아래 표 3과 같다.The administration method is shown in Table 3 below.
Figure PCTKR2021012692-appb-T000003
Figure PCTKR2021012692-appb-T000003
패혈증 유발 방법은, 시험물질 투여 후 30 분 후 (DAY 1)에 LPS (Lipopolysaccharide, Sigma Aldrich, Cat No. L3024)를 1 mL 주사기를 이용하여 복강 내에 10 mg/kg 용량으로 단회 투여하여 패혈증을 유발하였다.The sepsis induction method is to induce sepsis by administering LPS (Lipopolysaccharide, Sigma Aldrich, Cat No. L3024) once intraperitoneally at a dose of 10 mg/kg using a 1 mL syringe 30 minutes after administration of the test substance (DAY 1). did
평가방법은 하기와 같다. The evaluation method is as follows.
(1) 일반증상 : 투여 및 관찰기간 동안 사망여부, 일반증상의 종류, 발현일 및 증상의 정도를 1 일 1 회 관찰하고, 개체별로 기록하였다. 시험물질 투여 개시일을 DAY 0으로 설정하였다.(1) General symptoms: During the administration and observation period, the status of death, the type of general symptoms, the date of onset, and the severity of symptoms were observed once a day and recorded for each individual. The start date of administration of the test substance was set as DAY 0.
(2) 사이토카인(Cytokine) 측정 : LPS 투여 4 시간 후에 각 개체별로 복대정맥 채혈 후 혈액에서 분리한 혈청을 이용하여 TNF-alpha ELISA kit (R&D Systems®, Cat. No.: MTA00B)와 IL-6 ELISA kit (R&D Systems®, Cat. No.: M6000B)를 이용하여 분석하였다.(2) Cytokine measurement: TNF-alpha ELISA kit (R&D Systems ® , Cat. No.: MTA00B) and IL- using serum isolated from blood after abdominal vena cava blood collection for each individual 4 hours after LPS administration 6 ELISA kit (R&D Systems ® , Cat. No.: M6000B) was used for analysis.
(3) 통계분석 : 측정결과는 SPSS statistics 12.0K for medical science를 사용하였다. 모수적인 다중비교는 Student's t-test 또는 ONE-WAY ANOVA를 실시하였다. P<0.05인 경우 통계학적으로 유의하다고 판정하였다.(3) Statistical analysis: SPSS statistics 12.0K for medical science was used for measurement results. For parametric multiple comparisons, Student's t-test or ONE-WAY ANOVA was performed. When P<0.05, it was determined to be statistically significant.
실험결과는 하기와 같다.The experimental results are as follows.
(1) 일반증상 : DAY 1 이후 모든 군에서 웅크린 자세 (Crouching position), 쇠약 (Weakness), 떨림 (Shivering), 눈곱 (Eye discharge) 등이 관찰되었으며, 이는 LPS 투여에 따른 증상으로 확인된다. 시험물질에 의한 이상증상은 관찰되지 않았다. 따라서 수가용화 UDCA는 안전한 치료제가 될 수 있는 장점이 있다 할 것이다.(1) General symptoms: Crouching position, weakness, shivering, eye discharge, etc. were observed in all groups after DAY 1, which are confirmed as symptoms following LPS administration. No abnormal symptoms were observed due to the test substance. Therefore, it will be said that water-soluble UDCA has the advantage of being a safe therapeutic agent.
(2) 혈청 내 사이토카인 측정 결과 : 정상군 (G1)과 부형제 대조군 (G2) 및 음성대조군 (G3)을 비교하였을 때, LPS로 패혈증이 유발된 부형제 대조군 (G2)과 음성대조군 (G3)에서 TNF-alpha와 IL-6의 발현량이 모두 통계학적으로 유의하게 증가되었다 (P<0.01 or P<0.05). (2) Serum cytokine measurement results: When comparing the normal group (G1), the excipient control group (G2) and the negative control group (G3), in the excipient control group (G2) and negative control group (G3) induced by LPS sepsis Both TNF-alpha and IL-6 expression levels were significantly increased ( Po0.01 or Po0.05 ).
부형제 대조군 (G2)과 시험물질 150 mg/kg 투여군 (G5) 및 시험물질 450 mg/kg 투여군 (G6)을 비교하였을 때, TNF-alpha와 IL-6 모두 통계학적으로 유의한 감소가 관찰되었다 (P<0.01 or P<0.05). 즉 부형제 대조군(G2) 대비 시험물질(수가용화 UDCA)군은 TNF-alpha를 18.4~24.2% 억제하였고, IL-6는 35.1~35.4% 억제하였다.When comparing the excipient control group (G2), the test substance 150 mg/kg administration group (G5), and the test substance 450 mg/kg administration group (G6), a statistically significant decrease was observed for both TNF-alpha and IL-6 ( P < 0.01 or P < 0.05). That is, compared to the excipient control group (G2), the test substance (water-soluble UDCA) group inhibited TNF-alpha by 18.4 to 24.2%, and IL-6 by 35.1 to 35.4%.
음성대조군 (G3)과 양성대조물질 투여군 (G7 및 G8)을 비교하였을 때, TNF-alpha와 IL-6 수치가 통계학적으로 유의하게 감소되었다 (P<0.01). 따라서 LPS 유발 패혈증 동물의 사이토카인 억제 모델의 적격성이 증명되었다.When the negative control group (G3) and the positive control group (G7 and G8) were compared, the TNF-alpha and IL-6 levels were statistically significantly decreased ( Po0.01 ). Thus, the suitability of the cytokine inhibition model in animals with LPS-induced sepsis was demonstrated.
상기 시험물질(수가용화 UDCA)의 사이토카인 억제 효과는 도 7 및 도 8에 나타내었다.The cytokine inhibitory effect of the test substance (water-soluble UDCA) is shown in FIGS. 7 and 8 .
결론적으로 첫번째 수행한 본 시험은 마우스 패혈증 모델에서 시험물질(수가용화 UDCA)의 투여 농도에 따른 염증유발성 사이토카인(inflammatory cytokine) 감소에 대한 효과를 평가하기 위하여 수행하였다.In conclusion, this first test was performed to evaluate the effect on the reduction of inflammatory cytokines according to the administered concentration of the test substance (water-soluble UDCA) in a mouse sepsis model.
시험 결과, LPS 투여에 의하여 웅크린 자세, 떨림, 눈곱, 쇠약 증상이 나타난 것으로 보아, 패혈증 유발된 것으로 판단하였다. As a result of the test, it was judged that sepsis was induced, considering that a crouched posture, tremors, eye mucus, and weakness appeared by LPS administration.
특히, 혈청 내 염증성 사이토카인 측정 결과, 시험물질(수가용화 UDCA) 150 mg/kg 투여군과 450 mg/kg 투여군에서 TNF-alpha와 IL-6 모두에서 통계학적으로 유의한 감소가 관찰되었다.In particular, as a result of measuring inflammatory cytokines in serum, a statistically significant decrease was observed in both TNF-alpha and IL-6 in the 150 mg/kg and 450 mg/kg administration groups of the test substance (water-soluble UDCA).
따라서, 마우스 패혈증 모델에 시험물질(수가용화 UDCA)을 반복투여하였을 때, 초기 염증성 사이토카인 감소에 효력이 있었다.Therefore, when the test substance (water-soluble UDCA) was repeatedly administered to the mouse sepsis model, it was effective in reducing the initial inflammatory cytokines.
3-3. 내독소 유발 패혈증 동물모델에서 UDCA 정제 / 수가용화한 UDCA 제제 단독투여 시의 생존율 효과 평가3-3. Evaluation of the survival rate effect of UDCA tablets/solubilized UDCA formulation alone administered in an animal model of endotoxin-induced sepsis
흰색 파우더 형태의 결정형 UDCA 제제 혹은 수가용화 UDCA 제제의 단톡투여로 패혈증 및 급성 폐렴을 예방 및 치료할 수 있는 지를 비임상 실험으로 생존율 증가의 근거 자료로 확보하였다.Whether sepsis and acute pneumonia can be prevented and treated by single-tok administration of white powder crystalline UDCA preparations or water-soluble UDCA preparations were obtained as evidence of survival rate increase through non-clinical experiments.
시험군 구성은 부형제 대조군 (G1), 음성대조군 (G2), 시험물질(수가용화 UDCA) 50 mg/kg 투여군 (G3), 시험물질(수가용화 UDCA) 150 mg/kg 투여군 (G4), 시험물질(수가용화 UDCA) 450 mg/kg 투여군 (G5), UDCA (흰색 파우더 형태의 결정형 UDCA제제 :구입처 ㈜대웅바이오, 제조번호 B9007271) 투여군 (G6), 양성대조물질 경구투여군 (G7) 및 양성대조물질 복강투여군 (G8)으로 설정하였다. 시험군 당 마리 수는 생존율 평가군 (G1-G8)을 위해 8 마리로 설정하였다. 시험항목으로는 사망동물, 일반증상 관찰, 생존율 측정을 실시하였다. 상기 시험물질(수가용화 UDCA)의 단독투여시 생존율 효과는 표 4 및 도 9에 나타내었다. 표 4의 숫자는 생존 동물의 수(number)이다.The test group consists of excipient control group (G1), negative control group (G2), test substance (water soluble UDCA) 50 mg/kg administration group (G3), test substance (water solubilization UDCA) 150 mg/kg administration group (G4), test substance (Water-solubilized UDCA) 450 mg/kg administration group (G5), UDCA (crystalline UDCA formulation in white powder form: Where to buy Daewoong Bio, serial number B9007271) administration group (G6), positive control substance oral administration group (G7) and positive control substance It was set as the intraperitoneal administration group (G8). The number of animals per test group was set to 8 for the survival rate evaluation group (G1-G8). As test items, dead animals, observation of general symptoms, and survival rate were measured. The survival rate effect when the test substance (water-soluble UDCA) was administered alone is shown in Table 4 and FIG. 9 . The number in Table 4 is the number of surviving animals.
시험 결과는 다음과 같았다.The test results were as follows.
사망 개체 (생존율) 산출 결과, 부형제 대조군 (G1)에서 4 마리 (50.00%), 음성대조군 (G2)에서 5 마리 (37.50%), 시험물질 50 mg/kg 투여군 (G3)에서 6 마리 (25.00%), 150 mg/kg 투여군 (G4)에서 6 마리 (25.00%), 450 mg/kg 투여군 (G5)에서 7 마리 (12.50%), UDCA 투여군 (G6)에서 5 마리 (37.50%), 양성대조물질 경구투여군 (G7)에서 사망 개체가 없었으며 (100.00%), 양성대조물질 복강투여군 (G8)에서 1 마리 (87.50%) 사망하였다. As a result of calculating mortality (survival rate), 4 mice (50.00%) in the excipient control group (G1), 5 mice (37.50%) in the negative control group (G2), and 6 mice (25.00%) in the test substance 50 mg/kg administration group (G3) ), 6 mice (25.00%) in the 150 mg/kg administration group (G4), 7 mice (12.50%) in the 450 mg/kg administration group (G5), 5 mice (37.50%) in the UDCA administration group (G6), positive control There was no death in the oral administration group (G7) (100.00%), and one animal (87.50%) died in the intraperitoneal administration group (G8) of the positive control.
Figure PCTKR2021012692-appb-T000004
Figure PCTKR2021012692-appb-T000004
시험 결과, LPS 투여에 의하여 웅크린 자세, 떨림, 눈곱, 쇠약 증상이 나타난 것으로 보아, 패혈증 유발된 것으로 판단하였다. As a result of the test, it was judged that sepsis was induced, considering that a crouched posture, tremors, eye mucus, and weakness appeared by LPS administration.
그러나 생존율 확인 결과, 놀랍게도 결정형 UDCA 제제나 수가용화 UDCA 제제 단독투여에 의한 생존율의 증가는 전혀 관찰되지 않았다. 즉 G1 부형제 대조군의 생존율 50%나 G2 음성대조군의 생존율 37.5% 대비 시험물질(수가용화 UDCA)의 생존율은 오히려 더 낮아 12.5% ~ 25%만 보였다. 또한, 결정형 UDCA 제제 G6 투여군도 수가용화 UDCA 투여군 25% 보다는 향상된 37.5%를 나타냈으나 음성대조군의 생존율 수준을 보였을 뿐, G1 부형제 대조군 50% 보다는 더 낮았다. However, as a result of confirming the survival rate, surprisingly, no increase in the survival rate was observed at all by administration of the crystalline UDCA formulation or the water-soluble UDCA formulation alone. That is, compared to the 50% survival rate of the G1 excipient control group or the 37.5% survival rate of the G2 negative control group, the survival rate of the test substance (water-soluble UDCA) was rather lower, showing only 12.5% to 25%. In addition, the crystalline UDCA formulation G6 administration group also showed an improvement of 37.5% compared to 25% of the water-soluble UDCA administration group, but only showed the survival level of the negative control group, and was lower than the G1 vehicle control group 50%.
기존 논문에서 UDCA가 패혈증 치료에 도움을 줄 수 있다는 연구의 가설은 많이 발표가 되고 있으나, 직접 실험으로 증명해 발표된 연구 결과는 없다. 그러나, 본 발명가들은 실험을 통하여 놀랍게도 기존 UDCA 제제 형태인 정제나 캡슐제제 같은 결정형 UDCA 및 수가용화 UDCA 액상제제 등은 단독투여 시 내독소 LPS 유발 패혈증 동물모델의 생존율을 전혀 증가시키지 못함을 발견하였다. 오히려 투여하지 않는 것이 패혈증 치료에 더 도움이 될 수 있을 정도였다. In previous papers, many research hypotheses have been published that UDCA can help treat sepsis, but there are no published research results to prove it through direct experiments. However, the present inventors surprisingly found that, when administered alone, the survival rate of endotoxin LPS-induced sepsis animal model did not increase at all when crystalline UDCA and water-soluble UDCA liquid formulations such as tablets or capsules, which are conventional UDCA formulations, were administered alone. Rather, not administering it was to the extent that it could be more helpful in the treatment of sepsis.
따라서 본 발명자들은 상기 직접 수행한 LPS 유발 패혈증 모델실험 결과를 더 자세히 관찰한 결과, 시험물질(수가용화 UDCA)의 병용요법, 특히 양성대조물질로 사용한 항염증의 기본 처방제인 덱사메타손과의 병용요법을 고안하게 되었다. Therefore, the present inventors observed the results of the LPS-induced sepsis model experiment performed directly above in more detail. As a result, the combination therapy of the test substance (water-soluble UDCA), in particular, the combination therapy with dexamethasone, a basic anti-inflammatory drug used as a positive control, was used. came to devise
3-4. 내독소 유발 패혈증 동물모델에서 UDCA와 덱사메타손을 병용투여하기 위한 덱사메타손의 적정 용량 도출 3-4. Derivation of an appropriate dose of dexamethasone for co-administration of UDCA and dexamethasone in an animal model of endotoxin-induced sepsis
시험물질인 수가용화 UDCA를 덱사메타손과 병용하기 위하여, 먼저 저용량의 덱사메타손의 적정 용량을 도출하였다. In order to use the test substance, water-solubilized UDCA in combination with dexamethasone, an appropriate dose of low-dose dexamethasone was first derived.
먼저 LPS 유발 동물모델에서 덱사메타손 적정용량을 찾는 실험을 실시하였다.First, an experiment was conducted to find the appropriate dose of dexamethasone in an LPS-induced animal model.
양성대조물질 용량 결정을 위한 1 차 시험군 구성은 음성대조군 (G1), 양성대조물질 5 mg/kg 투여군 (G2), 1.5 mg/kg 투여군 (G3), 0.5 mg/kg 투여군 (G4), 0.15 mg/kg 투여군 (G5), 0.05 mg/kg 투여군 (G6)으로 설정하였다.The composition of the primary test group for determining the dose of the positive control material was the negative control group (G1), the positive control group administered with 5 mg/kg (G2), the group administered with 1.5 mg/kg (G3), the group administered with 0.5 mg/kg (G4), 0.15 A mg/kg administration group (G5) and a 0.05 mg/kg administration group (G6) were set.
양성대조물질 덱사메타손의 병용투여 용량 결정을 위한 1 차 시험의 경우, 음성대조군 (G1)에서 DAY 2 에 2 례, DAY 3 에 3 례 사망하였다. 양성대조물질 5 mg/kg 투여군 (G2)에서 DAY 3 에 1 례 사망하였고, 1.5 mg/kg 투여군 (G3)에서 DAY 3 에 3 례 사망하였으며, 0.5 mg/kg 투여군 (G4)에서 DAY 3 에 3 례 사망하였다. 양성대조물질 0.15 mg/kg 투여군 (G5)에서 DAY 2 에 2 례, DAY 3 에 4 례 사망하였고, 0.05 mg/kg 투여군 (G6)에서 DAY 2 에 2 례, DAY 3 에서 6 례 사망하였다. In the case of the primary test to determine the co-administration dose of the positive control substance, dexamethasone, 2 cases on DAY 2 and 3 cases on DAY 3 in the negative control group (G1) died. One case died on DAY 3 in the positive control group administered with 5 mg/kg (G2), 3 cases died on DAY 3 in the group administered with 1.5 mg/kg (G3), and 3 on DAY 3 in the group administered with 0.5 mg/kg (G4). case died. In the 0.15 mg/kg positive control group (G5), 2 cases died on DAY 2 and 4 cases on DAY 3, and 2 cases died on DAY 2 and 6 cases on DAY 3 in the 0.05 mg/kg administration group (G6).
생존율 평가 결과, 음성대조군 (G1)에서 37.50%, 양성대조물질 5 mg/kg 투여군 (G2)에서 87.50%, 양성대조물질 1.5 mg/kg 투여군 (G3)에서 62.50%, 양성대조물질 0.5 mg/kg 투여군 (G4)에서 62.50%, 양성대조물질 0.15 mg/kg 투여군 (G5)에서 25.00%, 양성대조물질 0.05 mg/kg 투여군 (G6)에서 0.00%의 생존율이 관찰되었다. As a result of the survival rate evaluation, 37.50% in the negative control group (G1), 87.50% in the 5 mg/kg positive control group (G2), 62.50% in the 1.5 mg/kg positive control group (G3), 0.5 mg/kg positive control material The survival rate was 62.50% in the administration group (G4), 25.00% in the 0.15 mg/kg positive control group (G5), and 0.00% in the positive control substance 0.05 mg/kg administration group (G6).
음성대조군 (G1)과 양성대조물질 5 mg/kg 투여군 (G2)을 비교하였을 때, 생존율이 통계학적으로 유의하였다 (P<0.05). When the negative control group (G1) and the 5 mg/kg positive control group (G2) were compared, the survival rate was statistically significant (P<0.05).
따라서, 음성대조군 (G1)의 생존율 37.5% 보다도 더 낮은 생존율을 보이고 0% 보다는 더 높은 G5군의 생존율을 바탕으로 양성대조물질의 병용 투여량을 0.15 mg/kg로 정하고, 시험물질과 병용 투여하여 생존율이 증가하는지 실시예 3-5 실험에서 확인하였다.Therefore, based on the survival rate of the negative control group (G1) lower than 37.5% and the survival rate of the G5 group higher than 0%, the combined dose of the positive control material was set at 0.15 mg/kg, and the Whether the survival rate is increased was confirmed in the experiments of Examples 3-5.
상기 시험 결과는 Kaplen-Meier survival analysis에 의해 통계학적으로 분석되었다. 그 결과를 표 5 및 도 10에 나타내었다. 표 5의 숫자는 생존 동물의 수(number)이다.The test results were statistically analyzed by Kaplen-Meier survival analysis. The results are shown in Table 5 and FIG. 10 . The number in Table 5 is the number of surviving animals.
Figure PCTKR2021012692-appb-T000005
Figure PCTKR2021012692-appb-T000005
3-5. 내독소 유발 패혈증 동물모델에서 UDCA 및 덱사메타손 병용투여 효과 3-5. Effect of combined administration of UDCA and dexamethasone in an animal model of endotoxin-induced sepsis
2 차 생존율 시험의 경우, 덱사메타손을 최저량을 사용하면서 시험물질(수가용화 UDCA)의 농도를 달리하면서 병용투여해 생존율 증가시키는지 확인하였다. In the case of the secondary survival rate test, it was confirmed whether dexamethasone was used in the lowest amount and administered in combination with different concentrations of the test substance (water-soluble UDCA) to increase the survival rate.
시험군 구성은 음성대조군 (G1), 양성대조물질 5 mg/kg 투여군 (G2), 양성대조물질 0.15 mg/kg 투여군 (G3), 부형제 및 양성대조물질 0.15 mg/kg 병용 투여군 (G4), 시험물질(수가용화 UDCA) 100 mg/kg 및 양성대조물질 0.15 mg/kg 병용 투여군 (G5), 시험물질(수가용화 UDCA) 200 mg/kg 및 양성대조물질 0.15 mg/kg 병용 투여군 (G6), 시험물질(수가용화 UDCA) 300 mg/kg 및 양성대조물질 0.15 mg/kg 병용 투여군 (G7), 시험물질 0.5 mg/head 비강투여군 (G8)으로 설정하였다. 시험군 당 마리 수는 모두 8 마리로 설정하였다.Test group consists of negative control group (G1), 5 mg/kg positive control group (G2), 0.15 mg/kg positive control group (G3), 0.15 mg/kg excipient and positive control substance combination group (G4), test Substance (water-solubilized UDCA) 100 mg/kg and positive control substance 0.15 mg/kg combined administration group (G5), test substance (water-solubilized UDCA) 200 mg/kg and positive control substance 0.15 mg/kg combination administration group (G6), test Substance (water-soluble UDCA) 300 mg/kg and positive control substance 0.15 mg/kg combined administration group (G7) and test substance 0.5 mg/head nasal administration group (G8) were set. The number of animals per test group was all set to 8.
2 차 시험 결과, 음성대조군 (G1)에서 DAY 2 에 1 례, DAY 3 에 5 례, DAY 4 에 1 례 사망하였다. 양성대조물질 5 mg/kg 투여군 (G2)에서 사망 개체가 없었고, 0.15 mg/kg 투여군 (G3)에서 DAY 3 에 5 례 사망하였다. 시험물질(수가용화 UDCA)-Vehicle과 양성대조물질 0.15 mg/kg 병용 투여군 (G4)에서 DAY 3 에 6 례 사망하였다. 시험물질(수가용화 UDCA) 100 mg/kg와 양성대조물질 0.15 mg/kg 병용 투여군 (G5)에서 DAY 3 에 2 례 사망하였다. 시험물질 (수가용화 UDCA) 200 mg/kg와 양성대조물질 0.15 mg/kg 병용 투여군 (G6)에서 DAY 3 에 2 례, DAY 5 에 1 례 사망하였다. 시험물질(수가용화 UDCA) 300 mg/kg와 양성대조물질 0.15 mg/kg 병용 투여군 (G7)에서 DAY 2 에 1 례, DAY 3 에 1 례, DAY 4 에 1 례 사망하였다. 시험물질(수가용화 UDCA) 0.5 mg/head/time 비강 투여군 (G8)에서 DAY 3 에 2 례 사망하였다. As a result of the second test, in the negative control group (G1), 1 case on DAY 2, 5 cases on DAY 3, and 1 case on DAY 4 died. There were no deaths in the 5 mg/kg positive control group (G2), and 5 deaths on DAY 3 in the 0.15 mg/kg administration group (G3). Six cases died on DAY 3 in the test substance (water-soluble UDCA)-vehicle and 0.15 mg/kg positive control group (G4). Two cases died on DAY 3 in the test substance (water-soluble UDCA) 100 mg/kg and positive control substance 0.15 mg/kg co-administered group (G5). In the test substance (water-soluble UDCA) 200 mg/kg and positive control substance 0.15 mg/kg group (G6), 2 cases died on DAY 3 and 1 case on DAY 5. In the test substance (water-soluble UDCA) 300 mg/kg and positive control substance 0.15 mg/kg group (G7), 1 case died on DAY 2, 1 case on DAY 3, and 1 case on DAY 4. Two cases died on DAY 3 in the test substance (water-soluble UDCA) 0.5 mg/head/time nasal administration group (G8).
따라서 생존율 평가 결과, 음성대조군 (G1)에서 12.50%, 양성대조물질 5 mg/kg 투여군 (G2)에서 100.00%, 양성대조물질 0.15 mg/kg 투여군 (G3)에서 37.50%, Vehicle과 양성대조물질 0.15 mg/kg 병용 투여군 (G4)에서 25.00%, 시험물질(수가용화 UDCA) 100 mg/kg와 양성대조물질 0.15 mg/kg 병용 투여군 (G5)에서 75.00%, 시험물질(수가용화 UDCA) 200 mg/kg와 양성대조물질 0.15 mg/kg 병용 투여군 (G6)에서 62.50%, 시험물질(수가용화 UDCA) 300mg/kg와 양성대조물질 0.15 mg/kg 병용 투여군 (G7)에서 62.50%, 시험물질(수가용화 UDCA) 0.5 mg/head/time 비강 투여군 (G8)에서 75.00%의 생존율이 관찰되었다. Therefore, as a result of the survival rate evaluation, 12.50% in the negative control group (G1), 100.00% in the 5 mg/kg positive control group (G2), 37.50% in the 0.15 mg/kg positive control group (G3), 0.15 vehicle and the positive control material In the mg/kg combination administration group (G4), 25.00%, test substance (water soluble UDCA) 100 mg/kg and positive control substance 0.15 mg/kg combination administration group (G5) 75.00%, test substance (water soluble UDCA) 200 mg/ 62.50% in the group (G6) administered in combination with kg and 0.15 mg/kg of the positive control, 62.50% in the group (G7) administered with the test substance (water-soluble UDCA) 300 mg/kg and the positive control at 0.15 mg/kg (G7) A survival rate of 75.00% was observed in the UDCA) 0.5 mg/head/time nasal administration group (G8).
음성대조군 (G1)과 양성대조물질 5 mg/kg 투여군 (G2), 시험물질(수가용화 UDCA) 100 mg/kg와 양성대조물질 0.15 mg/kg 병용 투여군 (G5), 시험물질(수가용화 UDCA) 200 mg/kg와 양성대조물질 0.15 mg/kg 병용 투여군 (G6), 및 시험물질(수가용화 UDCA) 0.5 mg/head/time 비강 투여군 (G8)을 비교하였을 때, 음성대조군 (G1)과 대비하여 모두 생존율이 통계학적으로 유의하였다 (P<0.01 or P<0.05).Negative control group (G1) and positive control substance 5 mg/kg administration group (G2), test substance (water-solubilized UDCA) 100 mg/kg and positive control substance 0.15 mg/kg group administration group (G5), test substance (water-solubilized UDCA) When comparing 200 mg/kg and 0.15 mg/kg positive control group (G6), and 0.5 mg/head/time nasal administration group (G8) of test substance (water-soluble UDCA), compared to negative control group (G1) All survival rates were statistically significant (P<0.01 or P<0.05).
결론적으로, 저용량 덱사메타손 0.15mg/kg 단독으로 투여시 생존율은 (G3)은 37.50%인데 반해 시험물질과 병용투여 한 경우 수가용화 UDCA가 100mg/kg 투여시(G5) 75.00%로, 놀랍게도 생존율이 200% (2.14배) 증가하였으며, 수가용화 UDCA가 200mg/kg 투여시(G6)에는 62.5%로 역시 166% (1.67배) 증가하였다.In conclusion, when administered with low-dose dexamethasone 0.15 mg/kg alone, the survival rate (G3) was 37.50%, whereas when administered in combination with a test substance, when water-solubilized UDCA was administered at 100 mg/kg (G5), the survival rate was 75.00%, surprisingly, the survival rate was 200 % (2.14 times) increased, and when the water-solubilized UDCA was administered at 200 mg/kg (G6), it was also increased by 166% (1.67 times) to 62.5%.
즉, 기존 결정형 UDCA 제제나 시험물질 수가용화 UDCA 제제 단독이나 저용량 덱사메타손 단독처리로는 도저히 생존율을 증가시킬 수 없었던 LPS 유발 패혈증 동물모델을 수가용화 UDCA와 저용량 덱사메카손을 병용할 시에는 생존율을 166% (1.67배) ~ 200% (2.14배) 까지 증가시켰기 때문에, 수가용화 UDCA는 덱사메타손과 병용투여시 패혈증 질환, ARDS 및 코로나바이러스감염증-19(Covid-19) 질환 치료에서 생존율을 높이는데 아주 귀중하게 사용될 수 있다. 특히 중증의 코로나바이러스감염증-19(Covid-19) 질환 치료시 저용량 덱사메타손을 짧은 기간밖에 처치하지 못하는데 수가용화 UDCA를 병용투여시 임상에서 생존율 증가에 영향을 줄 수 있는 근거자료가 될 수 있다. In other words, in an animal model of LPS-induced sepsis, which could not possibly increase the survival rate with the existing crystalline UDCA formulation or the test substance water-solubilized UDCA formulation alone or low-dose dexamethasone alone, the survival rate was 166 when water-solubilized UDCA and low-dose dexamecasone were used in combination. % (1.67-fold) to 200% (2.14-fold), solubilized UDCA is very valuable for increasing the survival rate in the treatment of sepsis disease, ARDS, and coronavirus disease-19 (Covid-19) disease when co-administered with dexamethasone can be used In particular, low-dose dexamethasone can only be treated for a short period of time in the treatment of severe coronavirus infection-19 (Covid-19) disease.
상기 2차 시험 결과를 표 6 및 도 11의 그래프로 나타내었다. 시험 결과는 Kaplen-Meier survival analysis에 의해 통계학적으로 분석된 것이고, 표 6의 숫자는 생존 동물의 수(number)이다.The results of the secondary test are shown in Table 6 and graphs of FIG. 11 . The test results were statistically analyzed by Kaplen-Meier survival analysis, and the number in Table 6 is the number of surviving animals.
Figure PCTKR2021012692-appb-T000006
Figure PCTKR2021012692-appb-T000006
상기 실시예와 같이 마우스 패혈증 모델에서 시험물질(수가용화 UDCA)의 단독 반복투여 및 병용 반복 투여에 따른 생존율 증가 효과를 평가해 본 바, LPS 투여에 의하여 웅크린 자세, 떨림, 눈곱 등의 증상이 나타난 것으로 보아, 패혈증 유발된 것으로 판단하였다. As in the above example, as in the mouse sepsis model, the effect of increasing the survival rate according to the repeated administration of the test substance (water-soluble UDCA) alone and the repeated administration in combination was evaluated. As a result, it was determined that sepsis was induced.
생존율 시험 결과, 시험물질인 수가용화 UDCA의 반복 투여 및 덱사메타손 같은 스테로이드를 저용량 사용하여 병용 반복 투여에 의하여 생존율 증가가 관찰되었다. 음성대조군과 비교하여 시험물질/양성대조물질 병용 투여군에서도 통계학적으로 유의한 차이가 관찰되었다. As a result of the survival rate test, it was observed that the survival rate was increased by repeated administration of the test substance, water-soluble UDCA, and the repeated administration of steroids such as dexamethasone at a low dose. Compared with the negative control group, a statistically significant difference was also observed in the test substance/positive control group administered together.
따라서, 마우스 패혈증 모델에 시험물질인 수가용화 UDCA는 반복 투여 또는 병용 투여하였을 때, 생존율 증가에 효과가 있는 것으로 판단된다. Therefore, it is judged that the test substance, water-soluble UDCA, is effective in increasing the survival rate when administered repeatedly or in combination in a mouse sepsis model.
결론적으로 마우스 패혈증 모델 시험을 통해서 시험물질(수가용화 UDCA)은 사이토카인 폭풍의 원인물질인 TNF-alpha와 IL-6의 발현량을 억제시키고, 저용량의 스테로이드인 덱사메타손과 병용투여시 생존율을 약 200%(2배) 가까이 증가시켰다. In conclusion, through the mouse sepsis model test, the test substance (water-soluble UDCA) suppressed the expression levels of TNF-alpha and IL-6, the causative agents of cytokine storm, and the survival rate when co-administered with dexamethasone, a low-dose steroid, was approximately 200 % (2 times) increased.
패혈증이 특히 급성 폐손상과 급성 호흡곤란증을 일으키는 원인으로 알려져 있으므로 수가용화 UDCA는 멕사메타손 같은 스테로이드와 병용할 경우 TNF-alpha 및 IL-6 같은 사이토카인 발현을 크게 억제시키고 생존율을 증가시킬 수 있다. 특히, 코로나바이러스감염증-19(covid-19)에서 나타나는 급성 폐손상으로 인한 급성 호흡곤란증 치료에서 생존율을 증가시킬 수 있다.Since sepsis is known to cause acute lung injury and acute respiratory distress, water-soluble UDCA significantly inhibits the expression of cytokines such as TNF-alpha and IL-6 and increases the survival rate when combined with steroids such as mexamethasone. . In particular, it can increase the survival rate in the treatment of acute respiratory distress caused by acute lung injury in COVID-19.
<실시예 4> 수가용화된 UDCA의 폐 조직내에 이행되는 약물동태학 분석<Example 4> Analysis of pharmacokinetics of solubilized UDCA in lung tissue
4-1. 위탁시험처4-1. Consignment test center
4-1-1) 약동학 동물실험 - 인제대학교 부산백병원/ 안과질환 T2B 기반 구축센터4-1-1) Pharmacokinetic Animal Experiment - Inje University Pusan Paik Hospital/ Ophthalmic Disease T2B Foundation Construction Center
4-1-2) 약동학 분석 - 인제대학교 약학대학4-1-2) Pharmacokinetic analysis - College of Pharmacy, Inje University
위탁시험처에 의뢰하여 수가용화된 UDCA 시험물질 (YSB201-4 혹은 AGP600이라고 명명)을 C57BL/6 마우스 모델에 경구투여한 후 시간에 따른 약물성분의 폐 조직 내에 이행되는 약동학적 추이를 분석하였다.After orally administering a water-solubilized UDCA test substance (named YSB201-4 or AGP600) to the C57BL/6 mouse model requested by a consignment laboratory, the pharmacokinetic transition of the drug component in the lung tissue according to time was analyzed.
4-2. 시험방법 및 재료4-2. Test method and material
4-2-1) 시험법4-2-1) Test method
(1) 시험물질을 마우스 모델에 경구투여한 후 각 시간별로 폐 조직 시료를 채취하고, 유기용매를 사용하여 생체 조직 내 약물 성분을 추출하여 그 농도를 HPLC 형광검출기를 통해 정량 분석하였다. 시료채취는 경구투여 후 0, 5, 10, 30분 후와 1, 2, 4, 10, 24, 48, 72시간 후에 실시하도록 하였으며, 각 시간마다 4마리 마우스에서 생체시료를 채취하였다. 시험군과 같은 절식과 식이를 한 대조군의 경우 0, 4, 10, 24, 48, 72시간 후 폐 조직의 생체시료를 채취하였다.(1) After oral administration of the test substance to the mouse model, lung tissue samples were collected for each hour, and drug components in living tissues were extracted using an organic solvent, and the concentration was quantitatively analyzed using an HPLC fluorescence detector. Sample collection was performed 0, 5, 10, 30 minutes and 1, 2, 4, 10, 24, 48, 72 hours after oral administration, and biological samples were collected from 4 mice at each hour. In the case of the control group that had fasted and fed the same as the test group, biological samples of lung tissue were collected after 0, 4, 10, 24, 48, and 72 hours.
(2) 유기용매를 사용하여 생체시료 내 시험물질 성분을 추출하고 그 회수율(recovery, %)을 계산함으로써 적합한 추출법을 정립하였다. 폐 조직에서 담즙산계열 성분을 특이적으로 정량 분석하기 위하여 효소반응과 형광 검출을 이용한 고속액체 크로마토그래피(HPLC)기기 시험법을 검증하였다.(2) An appropriate extraction method was established by extracting the component of a test substance in a biological sample using an organic solvent and calculating the recovery (%). In order to specifically quantitatively analyze bile acid-based components in lung tissue, a high-performance liquid chromatography (HPLC) test method using enzymatic reaction and fluorescence detection was verified.
(3) 폐 조직내 약물성분의 농도 분석결과를 약물동태분석 프로그램인 WinNonLin에 적용시켜 경구투여 후의 약동학 파라미터 (pharmacokinetic parameters)을 계산하고, AGP600 제제의 PK 프로파일을 확인하였다.(3) Pharmacokinetic parameters after oral administration were calculated by applying the concentration analysis results of drug components in lung tissue to WinNonLin, a pharmacokinetic analysis program, and the PK profile of the AGP600 formulation was confirmed.
(4) IACUC: 인제대학교 부산백병원 인당생명의학연구원 동물실험실은 2014년 식품의약품안전처로부터 인증을 획득하였으며, 본 시험계획서는 인제대학교 의과대학 IACUC(Institutional Animal Care and Use Committee) 심의를 통과하여 진행하였다(IACUC No. IJUBPH_2016-001-02).(4) IACUC: Inje University Pusan Paik Hospital In-Dang Biomedical Research Institute's animal laboratory obtained certification from the Ministry of Food and Drug Safety in 2014, and this test protocol was conducted after passing the Inje University School of Medicine IACUC (Institutional Animal Care and Use Committee) deliberation. (IACUC No. IJUBPH_2016-001-02).
4-2-2) 시험시약 및 장비4-2-2) Test reagents and equipment
(1) 시험물질로 AGP600(실시예 2의 수가용화된 UDCA 청정 수용액 원액; UDCA 농도: 25mg/ml)를 사용하였다. (1) AGP600 (undiluted aqueous solution of UDCA in Example 2; UDCA concentration: 25 mg/ml) was used as a test substance.
(2) 표준물질로 사용된 다양한 담즙산은 gUDCA(glycoursodeoxycholic acid), tUDCA(tauroursodeoxycholic acid), UDCA(ursodeoxycholic acid), GCA (Glycocholic acid hydrate), TCA(Taurocholic acid sodium salt hydrate), CA(Cholic acid), GCDCA(Glycochenodeoxycholic acid), TCDCA (Taurochenodeoxycholic acid), GDCA(Glycodeoxycholic acid), TDCA (Taurodeoxycholic acid), CDCA (Chenodeoxycholic acid), DCA (Deoxycholic acid), GLCA(Glycolithocholic acid sodium salt), TLCA(Taurolithocholic acid), 및 LCA(Lithocholic acid)로 미국 시그마-알드리치사(Sigma-Aldrich)에서 구매하였다.(2) Various bile acids used as standard materials are gUDCA (glycoursodeoxycholic acid), tUDCA (tauroursodeoxycholic acid), UDCA (ursodeoxycholic acid), GCA (Glycocholic acid hydrate), TCA (Taurocholic acid sodium salt hydrate), CA (Cholic acid) , GCDCA (Glycochenodeoxycholic acid), TCDCA (Taurochenodeoxycholic acid), GDCA (Glycodeoxycholic acid), TDCA (Taurodeoxycholic acid), CDCA (Chenodeoxycholic acid), DCA (Deoxycholic acid), GLCA (Glycolithocholic acid sodium salt), TLCA (Glycolithocholic acid sodium salt), TLCA , and LCA (Lithocholic acid) were purchased from Sigma-Aldrich in the United States.
(3) 분석장비는 미국 Water사의 2695 Alliance HPLC (high performance liquid chromatography) 기기이며, 일본 JASCO사의 BilePak II column (4.6125 mm, JASCO, Japan) 및 EnzymePak 3α-HSD column (4.635 mm, JASCO, Japan)을 함께 사용하였다.(3) The analysis equipment is 2695 Alliance HPLC (high performance liquid chromatography) equipment of Water Corporation of the United States, and BilePak II column (4.6125 mm, JASCO, Japan) and EnzymePak 3α-HSD column (4.635 mm, JASCO, Japan) of JASCO of Japan are used. used together.
4-2-3) 약물투여방법 - 경구투여4-2-3) Method of drug administration - Oral administration
경구투여 직전에 12시간 절식시킨 후 측정한 체중에 근거하여 개체별 투여액량을 환산하여 1회용 주사기에 존데를 장착하여 경구투여하였고 경구투여 후 4시간 이후부터는 고형사료를 재공급하였다.After fasting for 12 hours immediately before oral administration, the dose for each individual was converted based on the measured body weight, and a sonde was attached to a disposable syringe for oral administration.
4-2-4) 실험동물4-2-4) Experimental animals
시험계는 마우스 C57BL/6 36마리 (암컷)을 사용하였고, 체중범위 16~18 g의 마우스를 분양받아 7일간 순화한 후 실험에 사용하였다. 첫 투여시 전체 평균 체중의 ±20% 내외 동물을 사용하였다. 구입처는 (주)코아텍 (경기도 평택, 한국)이다. 동물실 환경은 온도 19~25℃, 습도 40~60%, 조도 150-300Lux로 맞추었고, 동물실과 사육 케이지(cage) 청소는 인제대학교 부산백병원 인당생명의학연구원의 표준작업지침서에 따라 실시하였다.For the test system, 36 mice C57BL/6 (females) were used, and mice having a body weight range of 16 to 18 g were received and acclimatized for 7 days before being used in the experiment. At the time of the first administration, animals within ±20% of the total average body weight were used. The place of purchase is Coretech Co., Ltd. (Pyeongtaek, Gyeonggi-do, Korea). The environment of the animal room was set at 19~25℃, 40~60% humidity, and 150-300Lux illuminance.
4-2-5) 시험군 구성 및 투여계획 4-2-5) Test group composition and administration plan
대조군 및 시험군의 구성 및 투여 계획은 표 7에 나타내었다.The composition and administration plan of the control and test groups are shown in Table 7.
Figure PCTKR2021012692-appb-T000007
Figure PCTKR2021012692-appb-T000007
4-2-6) 시료분석 방법4-2-6) Sample analysis method
마우스의 폐 조직 생체시료의 분석을 위해 일본 JASCO사의 담즙산 분석시스템을 이용하였다. Waters사의 Alliance HPLC시스템(Waters　2695 Alliance HPLC 시스템)을 구축하여 형광검출기(excitation: 345 nm, emission: 470 nm)를 통해 담즙산 물질의 농도를 정량 분석할 수 있도록 세팅하였고, HPLC 실험조건은 표 8과 같다.For the analysis of mouse lung tissue biological samples, the bile acid analysis system of JASCO of Japan was used. Waters' Alliance HPLC system (Waters　2695 Alliance HPLC system) was established to enable quantitative analysis of the concentration of bile acid substances through a fluorescence detector (excitation: 345 nm, emission: 470 nm), and the HPLC experimental conditions are shown in Table 8.
Figure PCTKR2021012692-appb-T000008
Figure PCTKR2021012692-appb-T000008
4-2-7) 자료 및 통계처리4-2-7) Data and statistical processing
시험결과를 Microsoft Excel 2010을 사용해 정리하고, 폐 조직내 농도 분석 결과에 대해 Pharsight WinNonlin 7.0 프로그램 (Certara, USA)을 사용해 추가적인 약동학적 분석을 수행하였다. GraphPad Prism 5.0을 사용하여 p<0.05 값을 통계적으로 유의한 값으로 보고 통계처리하였으며, 그래프를 작성하였다.The test results were summarized using Microsoft Excel 2010, and additional pharmacokinetic analysis was performed on the results of concentration analysis in lung tissue using the Pharsight WinNonlin 7.0 program (Certara, USA). Using GraphPad Prism 5.0, p<0.05 value was regarded as a statistically significant value and statistically processed, and a graph was prepared.
4-3. 폐(lung) 조직내로의 약동학 분석4-3. Pharmacokinetic analysis into lung tissue
목적: 시험물질 AGP600을 C57BL/6 마우스 모델에 경구투여한 후 시간에 따라 약물성분이 폐(lung)까지 치료적 활성량으로 운반되는지 약동학적 추이 분석을 실시하였다.Purpose: After oral administration of the test substance AGP600 to the C57BL/6 mouse model, a pharmacokinetic trend analysis was conducted to determine whether the drug component was transported in a therapeutically active amount to the lungs over time.
4-3-1) 약동학분석 결과4-3-1) Pharmacokinetic analysis results
4-3-1-1) 폐 조직 내 샘플 분석4-3-1-1) Sample analysis in lung tissue
도 12에서 보는 바와 같이, AGP600을 경구투여하면 수가용화된 UDCA가 빠르게 폐까지 전달되고, 또 폐에서 1시간 동안 일정 농도를 유지하였다. 이는 본 발명의 수가용화 UDCA 청정 수용액 제제가 경구투여만으로도 치료적 활성량으로 UDCA를 폐까지 운반시킬 수 있으며, 또한 운반된 UDCA를 바로 소실시키지 않고 1시간 이상 유지할 수 있음을 발견하게 되었다. 이어 UDCA의 대사물인 tUDCA는 도 13에서 보는 바와 같이, UDCA가 Tmax 이후 점차 소실되는 시간에 폐 조직 내에서 증가하여 일정 시간 유지되었다. UDCA 계열 담즙산인 UDCA 및 tUDCA (또는 gUDCA)는 세포 보호 기능(cytoprotection)을 하는 것으로 알려져 있다. 또한, 본 발명의 수가용화 UDCA가 경구투여한 경우, UDCA 계열 담즙산이 세포독성을 띠는 소수성 담즙산을 포함하는 비UDCA 계열의 다른 담즙산보다 높은 농도로 1시간 이상 폐 조직 내에 존재하는 것을 확인하였다(도 15).As shown in FIG. 12 , when AGP600 was orally administered, water-solubilized UDCA was rapidly delivered to the lungs, and a constant concentration was maintained in the lungs for 1 hour. It was found that the water-solubilized UDCA aqueous solution formulation of the present invention can transport UDCA in a therapeutically active amount to the lungs only by oral administration, and can maintain the delivered UDCA for 1 hour or more without immediately disappearing. Subsequently, tUDCA, a metabolite of UDCA, increased in the lung tissue at a time when UDCA gradually disappeared after Tmax and was maintained for a certain period of time, as shown in FIG. 13 . UDCA and tUDCA (or gUDCA), which are UDCA-family bile acids, are known to have a cytoprotection function. In addition, when the water-soluble UDCA of the present invention was orally administered, it was confirmed that the UDCA-based bile acids were present in the lung tissue at a higher concentration than other non-UDCA-based bile acids including cytotoxic hydrophobic bile acids for 1 hour or more ( 15).
AGP600 (경구투여량 125 mg/kg)을 C57BL/6 마우스 모델에 경구투여한 후 시간에 따라 약물성분이 폐(lung)까지 치료적 활성량으로 운반되는지 약동학적 추이를 분석을 실시하여 표 9, 표 19, 및 표 11에 나타내었다.After oral administration of AGP600 (oral dose: 125 mg/kg) to the C57BL/6 mouse model, the pharmacokinetic trend was analyzed to see if the drug component was transported in a therapeutically active amount to the lungs over time. Table 9, Table 19 and Table 11 are shown.
표 9 및 표 10에 나타난 바와 같이, 시험군 마우스 (1,2,3,4군) (n=4)의 폐(lung) 조직내 UDCA는 Tmax 0.083 시간에서 폐 조직 내 최고농도(Cmax)는 7.13 ± 2.29 ㎍/g tissue을 보였으며, AUC (hr·㎍/g tissue)는 12.79 ± 8.49이었다. UDCA 계열 담즙산 물질(UDCA, tUDCA 및 gUDCA)의 AUC 합계를 기타 담즙산 물질의 AUC 합계로 나눈 비율인 AUC ratio는 1.10 이었다. As shown in Tables 9 and 10, UDCA in lung tissue of test group mice ( groups 1,2,3,4) (n=4) was the highest concentration (Cmax) in lung tissue at Tmax 0.083 hours. It showed 7.13 ± 2.29 μg/g tissue, and AUC (hr·㎍/g tissue) was 12.79 ± 8.49. The AUC ratio, which is the ratio of the sum of AUCs of UDCA-family bile acids (UDCA, tUDCA, and gUDCA) divided by the sum of AUCs of other bile acids, was 1.10.
시간이 지남에 따라 UDCA의 생체내 대사체이자 세포보호 효과가 있는 tUDCA도 폐로 전달되었다. 폐(lung) 조직내 tUDCA는 Tmax 1.13 ± 0.31 시간에서 폐 조직 내 최고농도(Cmax) 4.36 ± 0.85 ㎍/g tissue 를 보였으며, AUC (hr·㎍/g tissue)는 28.31 ± 10.57 이었다. Over time, tUDCA, an in vivo metabolite of UDCA and a cytoprotective effect, was also delivered to the lungs. tUDCA in lung tissue showed the highest concentration (Cmax) in lung tissue (Cmax) of 4.36 ± 0.85 μg/g tissue at Tmax 1.13 ± 0.31 hours, and AUC (hr·㎍/g tissue) was 28.31 ± 10.57.
표 11에서와 같이 폐 조직에서 경구투여 후 1시간내 평균 UDCA 함량은 0.65 ± 1.03 ㎍/g tissue 이상 이었으며, 경구투여 후 2시간내 평균 TUDCA 함량은 0.13 ± 0.17 ㎍/g tissue 이었다.As shown in Table 11, the average UDCA content within 1 hour after oral administration in lung tissue was 0.65 ± 1.03 μg/g tissue or more, and the average TUDCA content within 2 hours after oral administration was 0.13 ± 0.17 μg/g tissue.
표 11 및 도 15에서 보는 바와 같이 폐 조직에서는 약물 투여 후 48시간 이후로 정량한계 이하의 농도로 나타났다. 그러나 수가용화되지 않은 UDCA 경우 비특허문헌 1(Masamichi Ota, et al., 1977)에서 확인할 수 있듯이 14C 방사능동위원소를 포함한 UDCA를 쥐(rat)에 30mg/Kg으로 1일 1회 간격으로 7일, 14일, 21일 반복투여 후 조직내 분포를 살펴보았는데 폐에서 방사성동위원소가 검출되지 않았다. 즉 수가용화되지 않은 UDCA는 경구복용 시 폐 조직내로의 UDCA를 거의 운반하지 못하는 것으로 확인하였다.As shown in Table 11 and FIG. 15 , concentrations below the limit of quantitation were found in the lung tissue after 48 hours after drug administration. However, in the case of non-solubilized UDCA, as can be seen in Non-Patent Document 1 (Masamichi Ota, et al., 1977), UDCA containing 14 C radioactive isotope was administered to rats at 30 mg/Kg at 30 mg/Kg once a day at intervals of 7 After repeated administration on days, 14, and 21 days, the distribution in tissues was examined, and no radioisotope was detected in the lungs. That is, it was confirmed that UDCA that is not solubilized hardly transports UDCA into the lung tissue when taken orally.
Figure PCTKR2021012692-appb-T000009
Figure PCTKR2021012692-appb-T000009
Figure PCTKR2021012692-appb-T000010
Figure PCTKR2021012692-appb-T000010
Figure PCTKR2021012692-appb-T000011
Figure PCTKR2021012692-appb-T000011
4-3-1-2) 폐 조직내 UDCA 계열 담즙산과 그 외 다른 담즙산들의 변화4-3-1-2) Changes in UDCA-type bile acids and other bile acids in lung tissue
도 13 내지 도 15를 참조하면, 상기와 같이 AGP600을 경구투여하면 수가용화된 UDCA가 빠르게 폐까지 전달되고, 또한 폐에서 1시간 동안 머무르면서 효과적으로 작용할 수 있었으며, UDCA 농도는 Tmax 이후 점차 소실되지만 이의 생체내 대사산물인 tUDCA가 다시 약 2시간 동안 운반돼 머물면서 세포보호 작용을 하였다. 따라서, 총 담즙산들이 폐 조직내 2시간 정도 머무르는 동안, 세포보호 기능(cytoprotection)을 하는 UDCA 계열 담즙산들(UDCA, TUDCA 및 GUDCA)의 농도의 합은 비UDCA 계열 담즙산들(예, TCA, CA)의 농도의 합보다도 항상 더 높았는데 본 발명의 수가용화 UDCA의 경구투여가 세포보호 기능이 뛰어난 UDCA 계열 담즙산들이 폐 조직내 일정 농도로 유지되도록 하여 폐 조직 세포를 더 효율적으로 보호할 수 있음을 확인하였다.13 to 15, when AGP600 is orally administered as described above, water-solubilized UDCA is rapidly delivered to the lungs, and it can act effectively while staying in the lungs for 1 hour. The UDCA concentration is gradually lost after Tmax, but its bio My metabolite, tUDCA, was transported again for about 2 hours and stayed there for cytoprotective action. Therefore, while total bile acids stay in the lung tissue for about 2 hours, the sum of the concentrations of UDCA-based bile acids (UDCA, TUDCA, and GUDCA) that have cytoprotection function is the sum of the concentrations of non-UDCA-based bile acids (eg, TCA, CA). It was always higher than the sum of the concentrations of did
따라서 본 발명의 수가용화 UDCA 경구투여는 먼저 UDCA가 높은 농도로 폐(lung) 조직까지 전달되고, 이후 스테로이드 일종인 덱사메타손을 병용 투여 시 덱사메타손 단독투여시 보다 더 효과적으로 손상된 폐세포의 기능을 회복시켜 생존율과 폐손상을 개선할 수 있음을 알 수 있다. Therefore, in the oral administration of water-soluble UDCA of the present invention, UDCA is first delivered to the lung tissue at a high concentration, and then, when dexamethasone, a type of steroid, is administered in combination, the function of damaged lung cells is restored more effectively than when dexamethasone is administered alone, resulting in survival rate. and lung damage can be improved.
특히 상기와 같은 결과는 수가용화 UDCA 계열 담즙산이 직접 폐 조직에서 손상된 폐세포의 보호작용을 나타내는 것으로, 박테리아에 의하거나 호흡기 바이러스인 인플루엔자 바이러스(influenza virus), 사스 코로나바이러스(SARS-CoV), 메르스 코로나바이러스(MERS-CoV), COVID-19 바이러스(SARS-CoV-2), 아데노바이러스(adenovirus), 파라인플루엔자 바이러스 (parainfluenza virus, PIV), RS 바이러스(respiratory syncytial virus, RSV), 라이노바이러스(rhinovirus)의 감염에 의하여 유발된 패혈증, 급성호흡곤란증후군 또는 폐손상에 직접 보호 효과를 나타낼 수 있는 것을 알 수 있다.In particular, the above results indicate that the water-soluble UDCA-based bile acid directly protects the damaged lung cells in the lung tissue, and is caused by bacteria or respiratory viruses such as influenza virus, SARS-CoV, and MER. MERS-CoV, COVID-19 virus (SARS-CoV-2), adenovirus, parainfluenza virus (PIV), respiratory syncytial virus (RSV), rhinovirus ( It can be seen that it can have a direct protective effect on sepsis, acute respiratory distress syndrome, or lung injury caused by infection with rhinovirus).
4-3-1-3) 약동학 분석 결론 및 고찰4-3-1-3) Pharmacokinetic analysis conclusion and consideration
UDCA를 폐 조직까지 높은 농도로 운반하기 위하여 UDCA를 수가용화시켜 보았는데, 본 발명자들은 놀랍게도 경구투여 만으로도 UDCA를 치료적 활성량으로 폐까지 운반시킬 수 있음을 알 수 있었다. 또한, 폐에서 UDCA는 바로 소실되지 않고 1시간 정도 머물러 효능을 발휘하고 순차적으로 UDCA의 대사물인 tUDCA까지 폐로 운반되 총 2시간 정도 머물면서 효능을 지속하였으므로 수가용화 UDCA에 의한 급성호흡곤란증후군 또는 폐손상의 예방 및 치료가 유효성이 있음이 밝혀졌다. In order to transport UDCA to the lung tissue at a high concentration, UDCA was solubilized. Surprisingly, the present inventors found that UDCA could be transported to the lungs in a therapeutically active amount only by oral administration. In addition, UDCA does not disappear from the lungs immediately and stays for about 1 hour to exert its efficacy, and tUDCA, a metabolite of UDCA, is sequentially transported to the lungs and stayed for a total of 2 hours. It has been found that the prevention and treatment of injuries are effective.
따라서 UDCA는 높은 농도로 폐(lung)까지 전달되고, 사이토카인 폭풍을 일으킨다고 알려진 초기 염증유발인자 TNF-a, IL-6를 억제하면서 동시에 효과적으로 손상된 폐세포의 기능을 회복시켜 생존율과 폐질환을 개선할 수 있음을 보여준다. Therefore, UDCA is delivered to the lungs at a high concentration and inhibits the early proinflammatory factors TNF-a and IL-6 known to cause a cytokine storm, while at the same time effectively restoring the function of damaged lung cells, thereby improving survival rate and lung disease. show that it can be improved.
본 발명자들은 간(liver) 치료에 주로 사용되는 담즙산의 일종인 우르소데옥시콜산(UDCA)이 박테리아나 호흡기 바이러스 등 감염로 인한 패혈증, 급성호흡곤란증 또는 급성 폐손상으로 발생하는 사이토카인 폭풍을 억제시키고, 생존율 증가 및 폐질환을 개선시킬 수 있는지 LPS-induced sepsis 동물모델 실험을 통해 확인해 보았다. 실험결과, 놀랍게도 UDCA를 경구투여 시 사이토카인 폭풍의 원인물질인 초기 염증유발인자 TNF-alpha와 IL-6의 과다 발현 억제효과가 있음을 발견하였다. 이에 더 나아가 생존율까지 증가시키는지 더 실험해 본 결과, 아주 실망스럽게도 UDCA 단독 경구투여만으로는 제제형태가 정제나 수가용화 제제에 상관없이 오히려 생존율을 증가시키지 못하는 것으로 확인되었다. 오히려 투여하지 않음만 보다 못하였다.The present inventors found that ursodeoxycholic acid (UDCA), a type of bile acid mainly used for liver treatment, suppresses cytokine storms caused by sepsis, acute respiratory distress or acute lung injury caused by infections such as bacteria or respiratory viruses, and , it was confirmed through LPS-induced sepsis animal model experiment whether it can improve the survival rate and lung disease. As a result of the experiment, surprisingly, it was found that oral administration of UDCA had an inhibitory effect on the overexpression of TNF-alpha and IL-6, which are the causative agents of the cytokine storm. Furthermore, as a result of further experimentation to see if it increases the survival rate, it was confirmed that, very disappointingly, the oral administration alone of UDCA did not increase the survival rate regardless of the tablet or water-solubilization formulation. Rather, it was worse than not administering.
이에 본 발명자들은 염증 억제 작용이 있는 합성 부신피질 호르몬 (코르티코스테로이드) 제제인 덱사메타손에 주목하였다. 즉 덱사메타손 제제가 LPS 유발 ㅍ패혈증 동물모델에서 생존율을 증가시키는지 직접 LPS 유발 패혈증 마우스 모델을 이용해 실험한 결과, 기존 논문 발표처럼 덱사메타손 제제는 고용량 반복 투여 때만 100% 생존율을 보였다, 그러나 놀랍게도 저용량 반복 투여 시에는 투여하지 않을 경우 0% 생존율 대비 겨우 36.5% 밖에 생존율을 증가시키지 못했다. LPS 유발 패혈증의 병 상태가 얼마나 중증이면 아무리 최고의 항염증 작용이 있다고 알려진 스테로이드이지만 낮은 용량 사용만으로는 생존율 100%를 달성하기 불가능함을 알 수 있었다. 한편, 덱사메타손 고용량 투여의 경우에는 100% 생존율을 보여 사람의 패혈증, 급성 폐손상 질환 및 급성 호흡곤란증 치료에 적용가능할 수도 있겠지만, 스테로이드를 사람에게서 고용량으로 장기간 사용 시 오히려 감염증 유발이나 감염증 악화 등의 부작용이 더 발생하는 치명적인 단점이 있다. 따라서 본 발명자들은 덱사메타손의 저용량 투여 방법을 치료제 개발에 활용할 것을 고안하였다. 왜냐하면, 덱사메타손을 저용량 투여시 비록 100% 생존율을 보이지는 않았지만 처리하지 않았을 때의 0% 생존율 대비해서는 36.5% 라는 생존율 증가를 보였기 때문이다. Accordingly, the present inventors paid attention to dexamethasone, a synthetic corticosteroid (corticosteroid) preparation having an anti-inflammatory action. In other words, as a result of a direct experiment using a mouse model of LPS-induced sepsis to see if dexamethasone increases the survival rate in an animal model of LPS-induced sepsis, dexamethasone showed a 100% survival rate only when high-dose repeated administration, as reported in previous papers. However, surprisingly, low-dose repeated administration When not administered, the survival rate was only increased by 36.5% compared to the 0% survival rate. No matter how severe the disease state of LPS-induced sepsis, it was found that it is impossible to achieve 100% survival rate by using a low dose only, although it is a steroid known to have the best anti-inflammatory action. On the other hand, high dose administration of dexamethasone shows a 100% survival rate, so it may be applicable to the treatment of sepsis, acute lung injury, and acute respiratory distress in humans. There are fatal drawbacks that occur further. Therefore, the present inventors devised to utilize a low-dose administration method of dexamethasone to develop a therapeutic agent. This is because, when dexamethasone was administered at a low dose, although it did not show a 100% survival rate, it showed an increase in the survival rate of 36.5% compared to the 0% survival rate when not treated.
따라서 본 발명자들은, 패혈증, 급성 폐손상 질환 및 급성 호흡곤란증 치료제 개발을 위하여 LPS-유발 패혈증 동물모델에서 UDCA와 덱사메타손의 저용량을 병용 반복 경구투여하는 것을 고안하게 되었다. Therefore, the present inventors have devised a combination of repeated oral administration of a low dose of UDCA and dexamethasone in an animal model of LPS-induced sepsis for the development of a therapeutic agent for sepsis, acute lung injury, and acute respiratory distress.
이에 본 발명자들은 UDCA와 스테로이드 일종인 덱사메타손의 저용량을 병용 반복 투여방식을 조합해 LPS 유발 패혈증 동물모델에 병용 투여 한 결과, 놀랍게도 UDCA 단독으로는 25.0~37.5% 대의 생존율에서 75%로 생존율이 증가하였고, 덱사메타손의 경우에도 덱사메타손 저용량 단독 투여 시의 생존율 37.5% 대비 수가용화 UDCA와 병용투여시 75% 라는 200%(2배) 이상 더 생존율을 증가시킬 수 있었다. 또한 동시에 체중을 회복시켰고, 폐 염증을 크게 개선시키는 것을 발견하였다.Accordingly, the present inventors combined UDCA and a low dose of dexamethasone, a steroid, in combination with a repeated administration method to administer LPS-induced sepsis in an animal model. Surprisingly, the survival rate of UDCA alone increased from 25.0 to 37.5% to 75% , In the case of dexamethasone, the survival rate could be increased by more than 200% (2 times) more than 37.5% when administered with a low dose of dexamethasone alone, 75% when administered in combination with water-soluble UDCA. It was also found that, at the same time, weight was restored and lung inflammation was greatly improved.
이상의 설명으로부터, 본 발명이 속하는 기술 분야의 당업자는 본 발명이 그 기술적 사상이나 필수적 특징을 변경하지 않고서 다른 구체적인 형태로 실시될 수 있다는 것을 이해할 수 있을 것이다. 이와 관련하여, 이상에서 기술한 실시 예들은 모든 면에서 예시적인 것이며 한정적인 것이 아닌 것으로서 이해해야만 한다. 본 발명의 범위는 상기 상세한 설명보다는 후술하는 특허 청구범위의 의미 및 범위 그리고 그 등가 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, all changes or modifications derived from the meaning and scope of the following claims and their equivalents.

Claims (21)

  1. 수가용화된 우르소데옥시콜산(Ursodeoxycholic acid, UDCA) 계열 담즙산 또는 이의 약학적으로 허용가능한 염을 포함하는 제1 제제; 및a first agent comprising a water-solubilized Ursodeoxycholic acid (UDCA)-based bile acid or a pharmaceutically acceptable salt thereof; and
    치료학적으로 허용가능한 항염증 제제 또는 항알러지 제제인 제2 제제;a second agent that is a therapeutically acceptable anti-inflammatory agent or an anti-allergic agent;
    를 포함하고, 염증성 사이토카인 발현을 억제하는, 패혈증 예방 또는 치료용, 약학 조성물.Including, inhibiting the expression of inflammatory cytokines, sepsis prevention or treatment for, pharmaceutical composition.
  2. 수가용화된 우르소데옥시콜산(Ursodeoxycholic acid) 계열 담즙산 또는 이의 약학적으로 허용가능한 염을 포함하는 제1 제제; 및a first agent comprising a water-solubilized Ursodeoxycholic acid-based bile acid or a pharmaceutically acceptable salt thereof; and
    치료학적으로 허용가능한 항염증 제제 또는 항알러지 제제인 제2 제제;a second agent that is a therapeutically acceptable anti-inflammatory agent or an anti-allergic agent;
    를 포함하고, 급성 호흡곤란증후군 또는 급성 폐손상 질환 예방 또는 치료용, 약학 조성물.Including, for preventing or treating acute respiratory distress syndrome or acute lung injury disease, a pharmaceutical composition.
  3. 제2항에 있어서, 3. The method of claim 2,
    급성 폐손상 질환 또는 급성 호흡곤란증후군은 세균 또는 호흡기 바이러스 감염에 의한 것인, 약학 조성물.Acute lung injury disease or acute respiratory distress syndrome is caused by a bacterial or respiratory viral infection, a pharmaceutical composition.
  4. 제3항에 있어서,4. The method of claim 3,
    상기 호흡기 바이러스는 인플루엔자 바이러스(influenza virus), 사스 코로나바이러스(SARS-CoV), 메르스 코로나바이러스(MERS-CoV), COVID-19 바이러스(SARS-CoV-2), 아데노바이러스(adenovirus), 파라인플루엔자 바이러스 (parainfluenza virus, PIV), RS 바이러스(respiratory syncytial virus, RSV), 및 라이노바이러스(rhinovirus) 중에서 선택된 1종 이상인 것을 특징으로 하는, 약학 조성물.The respiratory virus is influenza virus (influenza virus), SARS coronavirus (SARS-CoV), MERS coronavirus (MERS-CoV), COVID-19 virus (SARS-CoV-2), adenovirus (adenovirus), parainfluenza Virus (parainfluenza virus, PIV), RS virus (respiratory syncytial virus, RSV), and rhinovirus (rhinovirus), characterized in that at least one selected from, a pharmaceutical composition.
  5. 제2항에 있어서,3. The method of claim 2,
    상기 급성 폐손상 질환은 폐 염증 또는 폐 병변을 일으키고, 생존율을 감소시키는 것인, 약학 조성물.The acute lung injury disease causes lung inflammation or lung lesions, and reduces the survival rate, the pharmaceutical composition.
  6. 제1항 또는 제2항에 있어서,3. The method of claim 1 or 2,
    경구투여용 제제 형태인, 약학 조성물.A pharmaceutical composition in the form of a formulation for oral administration.
  7. 제1항 또는 제2항에 있어서,3. The method of claim 1 or 2,
    상기 수가용화된 UDCA 계열 담즙산은 경구투여 시 1일 5 내지 450mg/kg의 치료학적 유효량으로 투여되도록 포함되는, 약학 조성물.The water-solubilized UDCA-based bile acid is included to be administered in a therapeutically effective amount of 5 to 450 mg/kg per day when orally administered, a pharmaceutical composition.
  8. 제1항 또는 제2항에 있어서, 3. The method of claim 1 or 2,
    상기 약학 조성물은 5일 이상 그리고 1일 1회 이상 투여되는, 약학 조성물.The pharmaceutical composition is administered for at least 5 days and at least once a day, the pharmaceutical composition.
  9. 제1항 또는 제2항에 있어서,3. The method of claim 1 or 2,
    상기 수가용화된 UDCA 계열 담즙산은 수가용화된 우르소데옥시콜산(UDCA), 타우로우르소데옥시콜산(tUDCA) 및 글리코우르소데옥시콜산(gUDCA) 중에서 선택되는 1종 이상을 포함하는, 약학 조성물.The water-solubilized UDCA-based bile acid comprises at least one selected from water-solubilized ursodeoxycholic acid (UDCA), tauroursodeoxycholic acid (tUDCA) and glycoursodeoxycholic acid (gUDCA), a pharmaceutical composition.
  10. 제1항 또는 제2항에 있어서,3. The method of claim 1 or 2,
    상기 제1 제제는,The first agent is
    상기 UDCA 계열의 담즙산을 포함하는 제1 물질;a first material comprising the UDCA-based bile acid;
    말토덱스트린, 덱스트린, 액상 포도당, 옥수수 시럽 고체, 가용성 전분, 덱스트란 및 이들의 조합에서 선택되는 1종 이상의 수가용성 전분 전화물인 제 2 물질; 및a second substance which is an invert product of at least one water-soluble starch selected from maltodextrin, dextrin, liquid glucose, corn syrup solid, soluble starch, dextran, and combinations thereof; and
    물;을 포함하고,water; including;
    상기 제1 물질 및 상기 제2 물질이 모두, 선택된 pH 값 범위 내의 모든 pH 값에 대해 침전되지 않고 용액상태로 남아 있는, 약학 조성물. wherein both the first substance and the second substance remain in solution without precipitation for all pH values within the selected pH value range.
  11. 제10항에 있어서,11. The method of claim 10,
    상기 수가용성 전분 전화물은 말토덱스트린이고, The water-soluble starch inversion product is maltodextrin,
    상기 UDCA 계열의 담즙산에 대한 상기 말토덱스트린의 최소 중량비는 pH 값 6 내지 9에서 1:13 이상인, 약학 조성물.The minimum weight ratio of the maltodextrin to the UDCA-based bile acid is 1:13 or more at a pH value of 6 to 9, a pharmaceutical composition.
  12. 제10항에 있어서,11. The method of claim 10,
    상기 UDCA 계열의 담즙산은 제1 제제의 총 중량에 대하여 0.01 내지 5 중량부로 포함되고,The UDCA-based bile acid is included in an amount of 0.01 to 5 parts by weight based on the total weight of the first agent,
    상기 수가용화 전분 전화물은 제1 제제의 총 중량에 대하여 1 내지 70 중량부로 포함되는, 약학 조성물.The water-soluble starch inversion product is included in an amount of 1 to 70 parts by weight based on the total weight of the first formulation, a pharmaceutical composition.
  13. 제1항 또는 제2항에 있어서,3. The method of claim 1 or 2,
    약학조성물은 대상에 경구투여 시 폐(lung)에서 적어도 5 분 이상 UDCA, tUDCA 및 gUDCA를 포함하는 UDCA 계열의 담즙산의 농도의 합이 UDCA 계열이 아닌 담즙산의 농도의 합보다 높게 유지되도록 하는, 약학 조성물.When the pharmaceutical composition is orally administered to a subject, the sum of the concentrations of UDCA-based bile acids, including UDCA, tUDCA, and gUDCA, is maintained higher than the sum of the concentrations of non-UDCA-based bile acids in the lung for at least 5 minutes. composition.
  14. 제1항 또는 제2항에 있어서, 3. The method of claim 1 or 2,
    상기 UDCA 계열의 담즙산은 UDCA이고, 약학 조성물을 대상에 경구투여시 폐(lung) 조직 내에서 1시간 이내에 UDCA 농도가 0.65± 1.03 ㎍/g tissue 이상이 되도록 하는, 약학 조성물.The UDCA-based bile acid is UDCA, and when the pharmaceutical composition is orally administered to a subject, the UDCA concentration is 0.65±1.03 μg/g tissue or more within 1 hour in lung tissue, a pharmaceutical composition.
  15. 제1항 또는 제2항에 있어서, 3. The method of claim 1 or 2,
    상기 UDCA 계열의 담즙산은 tUDCA이고, 약학 조성물을 대상에 경구투여시 폐(lung) 조직 내에서 2시간 이내에 tUDCA 농도가 0.13 ± 0.17 ㎍/g tissue 이상 되도록 하는, 약학 조성물.The UDCA-based bile acid is tUDCA, and when the pharmaceutical composition is orally administered to a subject, the tUDCA concentration is 0.13 ± 0.17 μg/g tissue or more within 2 hours in lung tissue, a pharmaceutical composition.
  16. 제1항 또는 제2항에 있어서,3. The method of claim 1 or 2,
    상기 제1 제제는 액상, 시럽, 또는 건조된 제제 형태인, 약학 조성물.The first formulation is in the form of a liquid, syrup, or dried formulation, a pharmaceutical composition.
  17. 제1항 또는 제2항에 있어서,3. The method of claim 1 or 2,
    약학 조성물은 비강 또는 정맥 주사로 투여되는, 약학 조성물.The pharmaceutical composition is administered by nasal or intravenous injection.
  18. 제1항 또는 제2항에 있어서,3. The method of claim 1 or 2,
    상기 제2 제제는 스테로이드 제제 또는 이의 약학적으로 허용가능한 염을 포함하고, 생존율을 증가시키는, 약학 조성물.The second agent includes a steroid agent or a pharmaceutically acceptable salt thereof, and increases the survival rate, a pharmaceutical composition.
  19. 제18항에 있어서,19. The method of claim 18,
    상기 제2 제제는 당질코르티코이드계인, 약학 조성물.The second agent is a glucocorticoid type, pharmaceutical composition.
  20. 제18항에 있어서,19. The method of claim 18,
    상기 제2 제제는 1일 0.05 내지 0.10mg/kg/day의 치료학적 유효량으로 투여되도록 포함되는, 약학 조성물.The second agent is included to be administered in a therapeutically effective amount of 0.05 to 0.10 mg/kg/day per day, a pharmaceutical composition.
  21. 제18항에 있어서,19. The method of claim 18,
    제1 제제 및 제2 제제는 서로 독립적으로 투여되는, 약학 조성물.A pharmaceutical composition, wherein the first agent and the second agent are administered independently of each other.
PCT/KR2021/012692 2020-09-18 2021-09-16 Pharmaceutical composition for preventing or treating sepsis, acute lung injury, or acute respiratory distress syndrome, containing water-solubilized bile acid WO2022060121A1 (en)

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