Pharmaceutical compositions for controlled release of weakly acidic drugs and their use

Abstract

To provide a liposomal composition without initial burst release to reduce potential side effect and extend the therapeutic efficacy of a weak acid drug.SOLUTION: A pharmaceutical compositions contains at least one liposome. The liposome comprises: an external lipid bilayer including at least one vesicle-forming phospholipid and less than 15 mol% of sterol; and an internal aqueous medium including a weak acid drug and weak acid salt. The pharmaceutical compositions reduce the burst release of the weak acid drug. Also provided is the use of the pharmaceutical composition to treat respiratory diseases and reduce the side effect of the weak acid drug.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of U.S. Application No. 62 / 731,101, filed on September 14, 2018 the entire disclosure of which is incorporated herein by reference.

[0002] Field Disclosed herein is a pharmaceutical composition comprising at least one liposome encapsulating a weakly acidic drug, wherein a small amount of sterol in the external lipid bilayer of the liposome reduces or prevents burst release of the weakly acidic drug and / or maintains the release of the weakly acidic drug.

Background Art

[0003] Background Liposomes are microscopic structures composed of natural or synthetic lipid bilayers and function as reservoirs for therapeutic agents by forming internal compartments. Various liposome compositions have been designed as drug delivery vehicles with different sizes, permeabilities, and stabilities, and they are all designed to provide sustained drug release. However, these sustained - release liposome compositions generally exhibit a high initial burst of drug release increasing side effects during burst release and / or plasma drug levels outside the therapeutic range.

[0004] The release profile of a liposome composition depends on the structure of the liposome membrane and affects the performance of the liposome. Therefore, control of the release profile is an important prerequisite for effectively using liposomes as a drug delivery medium. For example, cholesterol in the external ​​​​​​​ When added to a lipid bilayer, it increases the rigidity and stability of the membrane and decreases the permeability of the lipid bilayer (S. Kaddah et al., Food Chem Toxicol. 2018 Mar;113:40-48). S. Kaddah et al. Release of capsule-encapsulated drugs increases cholesterol in the liposomal bilayer (up to 30%). This indicates a decrease in the associated value. E. Corvera et al. (Biochim Biophys Acta. 1992 Jun 30;1 107(2):261-70) shows that low concentrations of cholesterol (5~) are present in DMPC and DPPC liposomes. The addition of 8% suggests that liposome stability decreases and membrane permeability improves. .

[0005] To reduce potential side effects and extend the therapeutic effect of weakly acidic drugs, initial burstlili There is still a need for liposome compositions without ions. This invention addresses these and This addresses other needs. [Overview of the project]

[0006] Brief summary of the invention This invention relates to a pharmaceutical compound comprising one or more liposomes suspended in an external medium. The product is wherein the liposome is (a) at least one vesicle-forming phospholipid (vesicle-for The outer lipid bilayer (externa) contains ming phospholipid and less than 15 mol% sterols. (i) lipid bilayer, and (b) an internal aqueous medium containing a weakly acidic drug and a weakly acidic salt (int The pharmaceutical composition contains an ernal aqueous medium, and less than 65% by weight of the weakly acidic drug is used. The present invention provides a pharmaceutical composition that is released into the external medium within one hour of administration.

[0007] The present invention also comprises the step of administering a pharmaceutical composition described herein, for respiratory diseases. We will disclose the treatment methods.

[0008] Furthermore, an effective amount of the pharmaceutical composition described herein is used for patients who need to take a weakly acidic drug. The present invention provides a method for reducing the side effects of a weakly acidic drug, which involves a step of administering the drug to an elephant.

[0009] The terms "invention," "the invention," and "this invention" are used in this patent. The terms "s invention)" and "the present invention" are used in this patent and This is intended to broadly refer to all subject matter of the following claims, including these terms. The following statements do not limit the subject matter described herein or the meaning or scope of the claims. It should be understood that this does not limit the embodiments of the inventions included in this patent. The state is defined not by this summary, but by the following claims. This summary is the present invention This is a high-level overview of various aspects of the Ming dynasty, which will be further explained in the following detailed explanation section. This is an introduction to the concept of a "tsuka". This summary is important to the subject matter described in the claims or This is not intended to identify essential features, nor to define the scope of the subject matter described in the claims. It is not intended to be used alone to determine the scope. The subject matter is the entire specification. , as can be understood by referring to some or all of the drawings and the appropriate part of each claim. It should be done.

[0010] The present invention will become clearer upon reading the following drawings and detailed description. [Brief explanation of the drawing]

[0011] The detailed embodiments of the present invention will be described in detail below with reference to the following drawings. [Figure 1] FIG. 1 is a line graph showing the logarithm of the average plasma iloprost concentration in rats administered a liposome composition (LL021b3A2) containing iloprost, bicarbonate and HP-β-CD, a liposome composition (LL021m3A2) containing iloprost, bicarbonate and RM-β-CD, or an iloprost solution. [Figure 2] FIG. 2 is a line graph showing the ratio of the area under the plasma concentration-time curve (AUCt) from 0 hours to a specific time and the area under the plasma concentration-time curve (AUCinf) from 0 hours to infinity for a liposome composition (LL021b3A2) containing iloprost, bicarbonate and HP-β-CD, a liposome composition (LL021m3A2) containing iloprost, bicarbonate and RM-β-CD, or an iloprost solution.

Mode for Carrying Out the Invention

[0012] Detailed Description As used herein, the articles "a" and "an" refer to one or more of the grammatical objects of the article (i.e., at least one). By way of example, "an element" means one element or a plurality of elements.

[0013] All numbers are modified by the term "about". As used herein, the term "about" refers to a range of ±10% of a specific value.

[0014] The terms "comprise" or "comprising" generally mean that one or more features, ingredients or components may be present. It is used to mean "include" or "include."

[0015] The term "subject" refers to vertebrates with respiratory diseases or the treatment of respiratory diseases. This can refer to vertebrates that are considered to require medical treatment. Examples include mammals. This includes warm-blooded animals such as primates, but more preferably humans. Non-human primates are also included. The term "object" includes domestic animals such as cats and dogs, and livestock (for example, cows, horses, pigs, etc.). Sheep, goats, etc.) and laboratory animals (e.g., mice, rabbits, rats, gerbils, guinea pigs) This includes (such as swabs). Therefore, this specification includes veterinary uses and medical formulations. It will be done.

[0016] The term "treating" includes procedures for treatment and prophylacteries. c) Refers to both or preventive measures. Those who require treatment already For those suffering from respiratory disease or related disorder Elephants, animals prone to respiratory diseases or related disorders, or animals that need to be protected from respiratory diseases There is a target.

[0017] As used herein, weak acid drugs are defined unless otherwise specified or in context. Unless otherwise stated, this includes its pharmaceutically acceptable salts and their protonated forms. In this embodiment, the weakly acidic drug is a carboxyl group (-COOH), a hydroxyl group (-OH) A small number of groups selected from the group consisting of ), phosphate groups (-PO4) and any combination thereof. It contains at least one functional group. In other embodiments, the weakly acidic drug is 1 or more but less than about 7, and 2 or more. They have a pKa of less than approximately 6, 2 to 6.9, or 2.5 to 6. Furthermore, weakly acidic drugs are as described above. The carboxyl group (-COOH), hydroxyl group (-OH), and phosphate group (-P In addition to O4), it may contain one or more functional groups; such additional functional groups may be acidic to the drug. The degree of sexuality should not be significantly altered from the acidity of its non-functionalized counterpart. In one embodiment, a weakly acidic drug is used to treat pulmonary hypertension. In another embodiment, Acidic drugs include prostaglandins, prostacyclin receptor agonists, and glucocorticoids. These are coid or nonsteroidal anti-inflammatory drugs. Table 1 shows a non-limiting list of weakly acidic drugs of the present invention. Here is an example.

[0018] [Table 1-1]

[0019] [Table 1-2]

[0020] The terms "encapsulation" and "loaded" as used herein The terms "entrapped" and "Lipo" can be used interchangeably. Incorporation of biological activators (e.g., iloprost) into the aqueous medium inside the osome (inc It refers to an organization or association.

[0021] This disclosure relates to a pharmaceutical composition comprising one or more liposomes suspended in an external medium, the Liposomes contain (a) at least one vesicle-forming phospholipid and less than 15 mol% sterol (b) an external lipid bilayer containing a weakly acidic drug and a weakly acidic salt, and (b) an internal aqueous The medium contains, in which case less than 65% by weight of a weakly acidic drug is administered to the pharmaceutical composition for 1 hour. The present invention provides a pharmaceutical composition that is released into the external medium within a certain time.

[0022] In one specific embodiment, the sterols in the outer lipid bilayer are 15, 14, 13, 12 The amounts are less than 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, and 0.5 mol%. Other ingredients In a typical embodiment, the outer lipid bilayer is substantially sterol-free.

[0023] The encapsulation rate of weakly acidic drugs in pharmaceutical compositions is approximately 70%, 75%, or over 80%. .

[0024] The pharmaceutical composition reduces the burst release of encapsulated weakly acidic drugs. Reduce. In one embodiment, approximately 70%, 69%, 68%, 67%, and 66% of the weakly acidic drug. Less than 65% of the pharmaceutical composition is released within one hour of administration. As a result, the target area Side effects of weakly acidic drugs in certain areas (e.g., cough, sore throat, pharyngeal pain, nosebleeds, coughing up blood, and nausea) (Wheezing of the throat) in a pharmaceutical composition in which sterols in the outer lipid bilayer are 15 mol% or more. Compared to other methods, the amount is reduced. Furthermore, this pharmaceutical composition prolongs the release of weakly acidic drugs and reduces the frequency of administration. Rasu.

[0025] In one embodiment, the burst release of a weakly acidic drug from the disclosed pharmaceutical composition is internal Further reduction occurs with the addition or encapsulation of cyclodextrin in an aqueous medium. Non-limiting examples of clodextrins include α-CD, β-CD, γ-CD, and 2-hydro Xypropyl β-CD (HP-β-CD), sulfobutyl ether β-CD (SBE-β -CD), randomly methylated β-CD (RM-β-CD), or a combination thereof There is a combination. Preferably, the cyclodextrin is HP-β-CD, RM-β-CD or These are combinations of them. In one specific embodiment, a weak acid drug and cyclodextrin The molar ratios (drug / CD ratio) are approximately 0.06, 0.055, 0.05, 0.045, and 0. It is 0.4, 0.035, or less than or equal to 0.03.

[0026] Furthermore, the amount of sterols in the outer lipid bilayer is less than 15 mol%, as disclosed herein. A method for treating respiratory diseases includes the step of administering an effective amount of a pharmaceutical composition to a subject in need. A method for providing treatment is disclosed. The weak acid drug of the pharmaceutical composition disclosed herein Trilla is compared to pharmaceutical compositions having 15 mol% or more of sterols in the outer lipid bilayer. Compared to other conditions, the incidence decreases. Non-specific examples of respiratory diseases include pulmonary hypertension and interstitial lung disease. be.

[0027] Furthermore, the use of the pharmaceutical compositions disclosed herein for the treatment of respiratory diseases, or The use of the pharmaceutical compositions disclosed herein for the manufacture of drugs for the treatment of respiratory diseases It will be disclosed.

[0028] The present invention also provides a sterol in the outer lipid bilayer for subjects who need to take a weakly acidic drug. The pharmaceutical composition disclosed herein, in which the urea content is less than 15 mol%, is administered in an effective amount. This concerns methods for reducing the side effects of weakly acidic drugs.

[0029] Depending on the embodiment, the pharmaceutical composition disclosed herein is a weakly acidic drug in the upper respiratory tract. It is administered by inhalation to reduce side effects.

[0030] A. Liposome components As used herein, the term “liposome” refers to one or more liposomes that encapsulate an internal aqueous medium. Microscopic vesicles or particles composed of a lipid bilayer To form a liposome, at least one "vesicle-forming lipid" is required. The presence of "ipid" is necessary, which forms or is incorporated into the lipid bilayer. It is an amphiphilic lipid that can form liposomes. Using appropriate vesicle-forming lipids, liposomes are formed. It is possible to form a lipid bilayer. The vesicle-forming lipids are not limited to the following. However, phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidyl Dilinositol (PI), phosphatidic acid (PA), phosphatidylethanolamine Phospholipids such as phospholipids (PE) or phosphatidylserine (PS), and positively charged lipids These include charged lipids, such as lipids that are either pure or negatively charged.

[0031] The lipid bilayer of liposomes contains at least 1 vesicle-forming lipid and 0 (zero) mole% or more. It contains less than 15 mol% of sterols (e.g., 0 to 14.99 mol%). Here, the sterols Cholesterol is cholesterol, hexasuccinate cholesterol. A group consisting of te), ergosterol, lanosterol, and any combination thereof. Selected from, but not limited to, these. In specific embodiments, sterols are cholesterols. It is terror.

[0032] In some embodiments, the vesicle-forming lipid is a mixture of a first phospholipid and a second phospholipid. In certain embodiments, the first phospholipid is hydrogenated egg phosphatidylcholine (HEPC), hydrogenated soybean phosphatidylcholine (HSPC), dipalmitoyl phosphatidylcholine (DPPC), diste Aro Phosphatidylcholine (PC) is selected from the group consisting of ylphosphatidylcholine (DSPC), diarachidoylphosphatidylcholine, dimyristoylphosphatidylcholine (DMPC), egg phosphatidylcholine (EPC), soy phosphatidylcholine (SPC), oleoyl palmitoylphosphatidylcholine, dioleoylphosphatidylcholine (DOPC), dipetrocerinoylphosphatidylcholine, palmitoyleleidoylphosphatidylcholine, palmitoyloleoylphosphatidylcholine, dilauroylphosphatidylcholine (DLPC), diundecanoylphosphatidylcholine, didecanoylphosphatidylcholine, dinonanoylphosphatidylcholine, and any combination thereof. In other embodiments, the second phospholipid is a polyethylene glycol-modified phospholipid containing polyethylene glycol having a molecular weight of about 500 to about 10,000 daltons, such as 1,2-distearo-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000](DSPE-PEG2000), distearo-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000](DSPE-PEG2000). Aro These are negatively charged phospholipids such as ylphosphatidylglycerol (DSPG), dipalmitoylphosphatidylglycerol (DPPG), dimyristoylphosphatidylglycerol (DMPG), or dioleoylphosphatidylglycerol (DOPG). In specific embodiments, the molar ratio of the first phospholipid:cholesterol:second phospholipid is 75-99:0-14.9:0.1-25.

[0033] In other embodiments, the vesicle-forming lipid is a mixture of the first phospholipid and the charged lipid. In a specific embodiment, the vesicle-forming lipids are a first phospholipid, a second phospholipid, and a charged It is a mixture of lipids. The charged lipids include stearylamine and 1,2-dioleoyl-3. -Trimethylammonium-propane (DOTAP), 3β-[N-(N,N-dimethyl [aminoethane)-carbamoyl]cholesterol (DC-cholesterol), N 4 - This Steryl-spermine (GL67), Dimethyldioctadecylammonium (DDAB) , 1,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA ), ethylphosphocholine (ethyl PC), or combinations thereof. Other specific In the embodiment, the molar ratio of the first phospholipid:cholesterol:charged lipid is 75-99:0 The ratio is ~14.9:0.1~25.

[0034] In one embodiment, the molar percentage of HSPC, cholesterol, and DSPG in the lipid bilayer. The ratios are 75-99:0-14.9:0.1-25. In other embodiments, in the lipid bilayer... The molar percentages of HSPC, cholesterol, and DSPE-PEG2000 are 75-99:0 The ratio is ~14.9:0.1~25.

[0035] In one embodiment, the outer lipid bilayer of the liposome may further contain a surfactant. This is a nonionic surfactant, cationic surfactant, or zwitterionic surfactant. It is also acceptable. Nonionic surfactants do not have formally charged groups on their heads. On-ionic surfactants have a net positive charge at their heads. Zwitterionic surfactants have an electrical charge. It is neutral in general, but different atoms have formal positive and negative charges.

[0036] Non-limiting examples of nonionic surfactants include nonionic water-soluble mono-, di-, and nonionic water-soluble mono- and di-fatty acids of triglycerides and polyethylene glycol. Acid esters; nonionic water-soluble sorbitan fatty acid esters (e.g., TWEEN 20( Sorbitan (polyoxyethylene 20-sorbitan monooleate), SPAN 80, etc. Monooleate; nonionic water-soluble triblock copolymer (e.g., POLOXAME Poly(ethylene oxide) / poly- (Propylene oxide) / Poly(ethylene oxide) triblock copolymer) or There are derivatives.

[0037] A non-exclusive example of a cationic surfactant is dimethyldialkylammonium bromide. Alternatively, there is dodecyltrimethylammonium bromide.

[0038] A non-limiting example of an amphoteric surfactant is 3-(N,N-dimethylpalmityl There is ammonia-propanesulfonate.

[0039] According to the present invention, liposomes have a pH gradient between the internal aqueous medium and the external medium. To form the vesicles, they are prepared in a medium containing a weakly acidic salt. Vesicle-forming phospholipids and 15 When less than 1% of sterols come into contact with a medium containing a weakly acidic salt, a liposome suspension is formed. .

[0040] Liposomes in suspension are subjected to size reduction. The size of liposomes is usually... This refers to the diameter. Reducing the size of liposomes can be done by extrusion, sonication, homogenization techniques, or grinding. This can be achieved by many methods, including technology, which are well known and are of the skill of those skilled in the art. This can be carried out by extruding liposomes under pressure, with a specified pore size. This includes passing the material through a filter at least once. The filter is typically made of polycarbonate. However, it does not interact with liposomes and is strong enough to be extruded under sufficient pressure. They may be made from durable materials. The size of the liposomes can be reduced by sonication. This can be done. In ultrasonic treatment, sound wave energy is used to break or shear liposomes. Then, they will spontaneously reform into smaller liposomes. For example, sonication will A glass tube containing a posome suspension is placed in a sonic epicenter (sonic epicenter) generated by a bus-type sonicator. This can be done by immersing in the center, or by using a probe-type sonicator. It is permissible to do so, in which case the sound wave energy will be directed to the titanium in direct contact with the liposome suspension. It is generated by vibration of the probe. In this invention, liposomes are typically about 500 nm or less, approximately 400 nm or less, approximately 300 nm or less, approximately 200 nm or less, or approximately 100 nm The following have diameters ranging from approximately 50 nm to 500 nm.

[0041] After sizing, the concentration of the weakly acidic salt in the external medium is adjusted to control the relationship between the internal aqueous medium and the external medium. It provides a pH gradient in between. This is used, for example, in diafiltration, dialysis, ultrafiltration, and tangential flow filtration (tan). By methods such as (general flow filtration), the external medium is filtered into citrate buffer (H3C6H5 Do not replace it with a suitable buffer that does not contain weakly acidic salts such as O or phosphate buffer (H3PO4). It can be implemented in various ways.

[0042] Weakly acidic salts create a lower external and higher internal aqueous medium between the external and internal aqueous medium of liposomes. It provides an internal pH gradient. In one embodiment, the pH of the internal aqueous medium is lower than the pH of the external medium. It is at least 0.1 units higher. In other embodiments, the pH of the internal aqueous medium is the pH of the external medium. It is at least 1 unit higher than pH. In yet another embodiment, the pH of the internal aqueous medium is approximately 7 The pH is 8, 9, or 10, and the pH of the external medium is less than 7, less than 6, less than 5, less than 4, or less than 3. These are approximately 3-7, approximately 3.5-6.5, or approximately 4-6. In yet another exemplary embodiment... Therefore, the pH of the external medium exceeds the pKa of the weakly acidic drug.

[0043] Non-limiting examples of weakly acidic salts include carboxylates and bicarbonates.

[0044] As used herein, "bicarbonate salt" refers to a combination of bicarbonate anions and cationic compounds. This refers to a pharmaceutically acceptable salt compound containing an ON component. In one embodiment, the catecho of the salt compound is used. The main component is a metal. Non-specific examples of metals include potassium (K) and sodium (Na). ), calcium (Ca), magnesium (Mg), cesium (Cs), lithium (Li) Group IA or IIA metals such as iron (Fe) and nickel (Ni) or other Group IA or I There are metals other than those in Group IA. Examples of bicarbonates include, but are not limited to, potassium bicarbonate. M, sodium bicarbonate, calcium bicarbonate, magnesium bicarbonate, cesium bicarbonate, heavy carbon Lithium oxide, nickel bicarbonate, ferrous iron bicarbonate, or their Any combination is possible.

[0045] As used herein, "Carboxylic acid salt" is limited to the following: Although not, formate, acetate, propionate, butyrate, isobutyrate, valerate, isovalerate There are ammonium compounds or combinations thereof. In one exemplary embodiment, the acetate is sodium acetate. It is lium, calcium acetate, or a combination thereof.

[0046] The concentration of bicarbonate or carboxylate is 50 mM or higher, 100 mM or higher, or 150 mM or higher. Top, 200mM or more, 250mM or more, 300mM or more, 350mM or more, 400mM or more Above, 450mM and above, 500mM and above, 600mM and above, 700mM and above, 800mM and above 900mM, less than 1000mM, 50mM to less than 1000mM, 50mM to 800m M, 200mM to less than 1000mM, 200mM to 800mM, or 200mM to 6 00mM, 250mM to less than 1000mM, 250mM to 800mM, or 250m The ranges are M~600mM and 300mM~600mM.

[0047] The prepared liposomes are used for loading and administering weakly acidic drugs to the target. It can be stored for a considerable period of time. For example, liposomes are used for loading weakly acidic drugs (l It can be stored in the refrigerator for a considerable period before administration. Alternatively, before administration, liposomal The liposomes may be dehydrated, stored, then rehydrated, and loaded with a weakly acidic drug. Furthermore, dehydration may be performed after loading a weakly acidic drug. Dehydration is available and known in the art. It can be carried out by many methods. In some embodiments, lipos The material is dehydrated using a standard freeze-drying apparatus, i.e., dehydration under low-pressure conditions. Furthermore, liposomes may be frozen, for example, using liquid nitrogen. Before dehydration, saccharin Add the solution to the liposome environment, for example, a buffer containing liposomes, and the dehydrated liposomes Qualitative and integrity may be ensured. Examples of saccharides are limited to the following: However, maltose, lactose, sucrose, trehalose, dextrose, sorbitol These include cholitol, mannitol, xylitol, or combinations thereof.

[0048] As mentioned above, containing less than 15 mol% sterols or substantially sterol-free Prepare a liposome suspension for loading a weakly acidic drug. Specifically, a weakly acidic The sex drug is added to the external medium of the liposome, and the desired loading concentration and encapsulation efficiency are achieved. Pharmaceutical composition: Percentage of the amount of weakly acidic drug inside / encapsulated in the composition relative to the total amount of weakly acidic drug in the composition Incubate the resulting suspension until the solution is achieved, and then add the weakly acidic drug to the liposomal solution. Diffuse it into the aqueous medium inside the room.

[0049] B. Relationship between sterol content of the outer lipid bilayer and controlled release profile Less than 15 mol% (e.g., 0-14.99 mol%) of lipids in the outer lipid bilayer of liposomes The pharmaceutical composition of the present invention, which contains sterols, is a encapsulated weakly acidic drug called Bartholli. It reduces the acidity and therefore the side effects of weakly acidic drugs. Furthermore, it helps to achieve the desired therapeutic effect. A sufficient amount of weakly acidic drug is released from the pharmaceutical composition, and the release profile is liposome Release profile of a pharmaceutical composition having more than 15 mol% sterols in the outer lipid bilayer It is unexpectedly extended compared to [the previous period].

[0050] As used herein, the term "burst release" refers to a pharmaceutical compound. A weakly acidic drug encapsulated from the pharmaceutical composition within 1 hour (60 minutes) of administration of the product. Rapid and / or some rate exceeding 70, 69, 68, 67, 66 or 65% This refers to an uncontrolled release.

[0051] As used herein, the term "extended release" is used in relation to "controlled release." "controlled release", "delayed release", "modified release" )", "prolonged release", "programmed release", "Time release," "rate control," or "sustained release" It can be used interchangeably with "lease" and within 1 hour after administering the pharmaceutical composition, a weak acid This refers to the release of less than 50%, 45%, or 40% of sex drugs.

[0052] In one embodiment, the burst-release or sustained-release profile of the pharmaceutical composition is captured. In vitro release (IVR) assays and / or in vivo pharmacokinetics of weakly acidic drugs. Based on research.

[0053] In certain embodiments, in vitro release (IVR) assays and / or in vivo drug assays are performed. Based on kinetic studies, the pharmaceutical composition contains approximately 70, 69, 68, and 67% of the captured weakly acidic drug. If less than 66 or 65% by weight of the pharmaceutical composition is released within one hour from the time of administration... It has the following emission profile.

[0054] C. Administration The pharmaceutical composition of the present invention can be administered to a cavity of an object that does not come into direct contact with blood. Examples of routes include, but are not limited to, inhalation, intratracheal injection, subcutaneous injection, intra-articular injection, There are intramuscular injections, intravitreous injections, and meningeal injections.

[0055] The pharmaceutical composition of the present invention may also be administered directly to the blood of the subject.

[0056] According to this disclosure, the pharmaceutical composition is taken once to three times a day, once every two days, or every three days. It may be administered once per day.

[0057] This disclosure is further illustrated by the following examples. However, the following examples are for illustrative purposes only. It is understood that this disclosure should not be interpreted as actually limiting the scope of this disclosure. sea ​​bream.

[0058] Examples General experimental procedure: 1. Preparation of Iloprost liposome composition Liposome colloid suspensions were prepared using ethanol injection technology. 98:2 or The first phospholipid (HSPC) and the second phospholipid (DSPE-) are mixed in a molar ratio of 98.5:1.5. All lipid components, including PEG2000 or DSPG, are stored in 2.86 mL at approximately 60°C. It was dissolved in an ethanol solution. The resulting lipid solution was then mixed with (2-hydroxypropyl) if necessary. 17.4 mL containing β-cyclodextrin (i.e., 45-120 mM) Inject into a sodium bicarbonate solution (100-400 mM; pH 8.5) and then into the liposome water. The mixture was mixed vigorously at 60°C for merging. The mixture was then sieved to a pore size of 0.2 or 0.1 μm. Extruded 6-10 times through a polycarbonate film of m thickness, in the range of approximately 100nm-200nm. Suspension of liposomes having an average particle size and a polydispersity index (PdI) of <0.2 A solution was obtained. The liposome suspension was then mixed with 10 mM sodium citrate buffer (pH 5.5). ) is dialyzed using a tangential flow filtration system, and lipo A transmembrane pH gradient is created between the internal aqueous medium and the external medium of the som. A pH gradient was formed (i.e., the inside was higher and the outside was lower). Next, A liposome suspension with a pH gradient is used in a drug loading process. It was stored at 4°C until the end.

[0059] Iloprost (purchased from Cayman Chemical, USA) in a 50 mM sodium citrate solution Dissolve in [a solution] and add to the liposome suspension so that the drug concentration is 1000-250 μg / mL. The mixture was added and incubated at 37°C for 30 minutes. The resulting product was buffered with sodium citrate. The pH was adjusted with a solution (pH 5.5), and the pH in the external medium was 5.5 in the liposome suspension. A liposome composition loaded with iloprost, which has a phospholipid concentration of 10 mM (iloprost- A loaded liposomal composition was obtained.

[0060] 2. Preparation of Ambrisentan liposome composition With or without (2-hydroxypropyl)-β-cyclodextrin, A liposome suspension was prepared according to step 1 above. Ambrisentan (Cayman Chemistry) After dissolving the drug (purchased from mical, USA) in dimethyl sulfoxide (DMSO), the specified drug concentration is used. Add the solution to the liposome suspension to a concentration of approximately 500 μg / mL and incubate at 37°C for 30 minutes. The mixture was incubated. The resulting product was adjusted with sodium citrate buffer (pH 5.5). Furthermore, the pH of the external medium is 5.5, and the phospholipid concentration in the liposome suspension is 10 mM. Ambrisentan-loaded liposomal (Composition) was obtained.

[0061] 3. Quantitative Characterization of Liposome Compositions a. Concentration of encapsulated free iloprost / ambrisentan iloprost or ambrisentan liposome composition in PD MiniTrap T M The encapsulated drug is injected into a G-25 column (GE Healthcare) and separated from the free drug. Released. Mix the iloprost or ambrisentan liposome composition with methanol. (90% methanol by volume and 10% liposome suspension), liposome-methylethanol A mixture was formed.

[0062] The concentrations of encapsulated iloprost and free iloprost are measured in the photodiode. A 30 μL liposome sample was placed in a Waters Acquity HPLC system equipped with a Ray (PDA) detector. Analysis was performed by injecting a methanol mixture. The mobile phase was acetonitrile, methanol The mobile phase is a mixture of phosphate buffer (pH 2.5) in a volume ratio of 36:17:47. The flow rate is 1.0 mL / min. Separation was performed at 25°C on a 3.9 mm × 15.0 cm, 5 The procedure was performed using a 0.0 μm C8 column, and the absorbance peak was detected at 205 nm.

[0063] The concentrations of encapsulated ambrisentan and free ambrisentan are, by mass A 1 μL liposome-methyl methyl ion was added to the Waters Acquity UPLC system equipped with a detector (QDa). Analysis was performed by injecting a mixture. Mobile phase A was acetonitrile with 0.1% formic acid. The mobile phase B contained ddH2O with 0.1% formic acid. The gradient conditions were as follows: It was: 50% mobile phase A for 0.2 minutes, 10% mobile phase A by 2 minutes, and 5 by 5.5 minutes. 0% mobile phase A. Separation was performed using a C18 column with dimensions of 4.6 mm × 10.0 cm and a particle size of 3.0 μm. The procedure was performed at 35°C with a flow rate of 1.0 mL / min. MS acquisition was performed as follows: Ambrisentan [M+H] + Ion, using SIR mode with m / z 347.2 I went there.

[0064] b. Encapsulation efficiency (EE) and drug-to-cyclodextrin ratio (drug-to-cyclodextrin ratio) (ratio of n): The total concentration of the drug (iloprost or ambrisentan) in the liposome composition is The mixture contains an encapsulated drug (L) in an internal aqueous medium and a free drug (F) in an external medium. Born.

[0065] Drug encapsulation efficiency (EE) is the ratio of the internal water content of liposomes to the total amount of drug (L+F). The following formula was used to calculate the percentage (%) of encapsulated drug (L) in the sexual medium. thing:

[0066]

number

[0067] ILO / CD ratio and ambrisentan liposome group of iloprostriplisome composition The AMB / CD ratio of the product was calculated using the following formula:

[0068]

number

[0069] c. Average particle size and polydispersity index (PdI): The average particle size of liposomes was evaluated by dynamic light scattering. The polydispersity index (PdI), a value indicating the fabric, is used for Beckman Coulter Delsa™ Nano C. The average particle size was measured using a tickle analyzer and the same evaluation method.

[0070] Example 1: In vitro release of iloprostriposome compositions with different amounts of sterols ( IVR Profile A. In vitro release (IVR) assay The iloprost liposome composition is formulated, and the iloprost concentration is determined by the general experimental method described above. The analysis was performed according to the procedure in the next section. The average particle size of liposomes was 100-200. The value was nm, and the PdI was less than 0.20.

[0071] Various IVR assays can be used to evaluate IVR profiles. Issey operates in accordance with iloprost in the liposome composition described in the claims. This is known to those skilled in the art, or will be obvious to those skilled in the art. Iloprost release from liposomes The output profile was obtained using simulated lung fluid (SLF) at 37°C with a shaking rate of 100 rpm [Dissolution T [echnologies 2011, 18, 15-28] iloprost with a starting phospholipid concentration of 10 mM Obtained by a 10-fold dilution of the liposome solution loaded with ilopros. Ilopros released at each time point. The percentage of releases (Release %) is calculated using the following formula, and the incubation period is calculated at a specific point in time (T). By comparing the encapsulation efficiency (EE) after encapsulation with the initial (T0) encapsulation efficiency, That's what I calculated.

[0072]

number

[0073] result: The physicochemical properties and IVR profiles of iloprostriposome compositions with different sterol levels are shown. 2 This will be shown.

[0074] [Table 2]

[0075] table 2 Therefore, it is shown that using sodium bicarbonate salts resulted in an EE of >90%, and that iloprost liposome compositions containing less than 15 mol% cholesterol released less than 65% of iloprost within 1 hour of SLF incubation, while iloprost liposome compositions containing 15 mol% or more cholesterol released more than 70% of iloprost within 1 hour of SLF incubation at 37°C.

[0076] Example 2: In vitro release of ambrisentan liposome compositions with different amounts of sterols Interventional Radiology (IVR) Profile The ambrisentan liposome composition is formulated, and the concentration of ambrisentan is the same as the general The analysis was performed according to the procedure in the experimental procedure section. The average particle size of liposomes was 100. The wavelength was approximately 200 nm, and the PdI value was less than 0.20.

[0077] result: The physicochemical properties and IVR profiles of ambrisentan liposome compositions with different sterol levels are shown. 3 This will be shown.

[0078] [Table 3]

[0079] table 3 Therefore, it is shown that using sodium bicarbonate salts achieves an EE of >90%, and that an ambrisentan liposome composition containing less than 15 mol% cholesterol released less than 50% of ambrisentan within 1 hour of SLF incubation at 37°C.

[0080] Example 3: Iloprostriposomes containing or not containing cyclodextrin (CD) In vitro release (IVR) profile of the composition Release profile of iloprost liposome composition of Example 1 within the liposome Cyclodextrin ((2-hydroxypropyl)-β-cyclodextrin) in aqueous media An in vitro study was conducted to evaluate the effects of phosphorus (HP-β-CD).

[0081] result: The physicochemical properties and IVR profiles of iloprostriposome compositions containing or not containing cyclodextrin (HP-β-CD) are shown. 4 This will be shown.

[0082] [Table 4]

[0083] table 4 This indicates that the addition of cyclodextrin further reduces burst release (less than 60% of iloprost is released within 1 hour of SLF incubation at 37°C), while maintaining the release properties of the iloprost liposome composition (less than 40% of iloprost is released within 1 hour of SLF incubation at 37°C).

[0084] Example 4: Encapsulation efficiency of iloprostriposome compositions using different weakly acidic salts Different weakly acidic salts affect the encapsulation efficiency of the iloprostriposome composition of Example 1. In vitro studies were conducted to evaluate the effects. In this example, a sodium bicarbonate solution was used. Iloprost was loaded using (400 mM) and sodium acetate solution.

[0085] result: The encapsulation efficiency of iloprostriposome compositions using different weakly acidic salts is shown. 5 This will be shown.

[0086] [Table 5]

[0087] table 5 Therefore, it is shown that using bicarbonates and acetates achieves an EE of >80%, and the presence of cyclodextrin in the internal aqueous medium further reduces burst release and sustains iloprost release from the liposome composition.

[0088] Example 5: Ilopros with different iloprosto-cyclodextrin (ILO / CD) ratios In vitro release (IVR) profile and in vivo pharmacokinetics of triposome compositions PK) Parameters Different ILO / CD ratios for Iloprostriposome compositions relative to IVR profiles To evaluate the effects, an in vitro study was conducted. Following the procedure outlined in Example 1, The liposome composition used in this study was prepared, and its IVR profile was analyzed. (Iloprost solution) (20 μg / mL) is added to a 2 mM tromethamine solution adjusted to a pH of approximately 8.4. It was prepared by dissolving the lost material.

[0089] B. In vivo pharmacokinetic (PK) studies of iloprostriposome compositions In this in vivo PK study, three male Sprague-Dawley rats (purchased from BioLASCO Taiwan Co., Ltd.) from each group were anesthetized with isoflurane and hooked around the maxillary incisors. The rats were placed in a supine position on an arched platform with their backs firmly supported, using a ribbon to support their backs. A microspray aerosol tip (Microsprayer, PennCentury, Philadelphia, USA) was inserted into the tracheal bifurcation of each rat, and the test sample (i.e., the surface) was inserted. 6 The composition (or iloprost solution) was administered intratracheally to each rat at a predetermined dose of 60 μg / kg using a high-pressure syringe attached to a microspray aerosol device.

[0090] Predetermined time points (i.e., 5, 30 minutes, 1.5, 3, 6, 7, and 8 hours after administration) Then, blood samples were collected from each rat into heparin-coated tubes and placed on wet ice. Placed. Next, the blood sample was subjected to a 4±2 oz test at approximately 2500 × g for 15 minutes within 1 hour of collection. The plasma was separated from the blood cells by centrifugation at °C. Approximately 0.1 mL of plasma sample from each rat was then prepared. In addition to being stored in a storage tube, it was stored at -70±2℃.

[0091] To measure plasma iloprost concentration, a 50 μL plasma sample was placed in a 96-well plate. After transferring to the wells, 150 μL of acetonitrile was added to each well. The resulting mixture The mixture is vortexed for 1 minute to break the binding of plasma proteins to iloprost, followed by The mixture was centrifuged at 3000 rpm for 5 minutes. The supernatant (150 μL) was mixed with an equal volume of H2O. Analysis was performed using liquid chromatography-tandem mass spectrometry (LC-MS / MS) to determine the rats' characteristics. Plasma iloprost concentration was measured.

[0092] result: IVR profiles and PK parameters (C) of iloprostriposome compositions with different ILO / CD ratios max ) 6 This is shown in Figures 1 and 2.

[0093] [Table 6]

[0094] table 6 From this, iloprost-liposome compositions with an ILO / CD ratio of less than 0.06 showed a reduced burst release profile (less than 68.7% of iloprost was released within 1 hour of administration). A more sustained release attribute (less than 45% of iloprost was released within 1 hour of SLF incubation at 37°C) was observed in iloprost-liposome compositions with an ILO / CD ratio of less than 0.026. A similar trend was observed when cyclodextrin was added to the internal aqueous medium.

[0095] Figure 1 shows the table at a given dose. 7 This shows the logarithm of mean plasma iloprost concentrations versus administration time up to 24 hours in rats administered with the iloprost-liposome composition (LL021b3A2 / LL021m3A2) or iloprost solution. While there is a peak within 1 hour of administration of the iloprost solution, there is no significant peak after administration of the iloprost-liposome composition. The reduction in peak release helps prevent drug side effects, for example, less local irritation in the upper respiratory tract upon direct contact with the liposome composition as defined in the claims.

[0096] Figure 2 shows how to determine the total exposure to iloprost over a certain period and for each composition (Table 7 To normalize different doses of iloprost in iloprost-liposome compositions or iloprost solutions, the area under the plasma concentration-time curve (AUC) from zero to a specific time is used. t) and the area under the plasma concentration-time curve (AUC) from zero time to infinite time. inf The ratio is shown. Compared to 100% iloprost being released within 1 hour of administration of iloprost solution, more than 80% of iloprost was released within 24 hours of administration of the iloprost-liposome composition. These results indicate reduced drug accumulation at the target site and therefore fewer side effects.

[0097] Example 6: Iloprostriposome composition containing different cyclodextrins (CDs) In vitro release (IVR) profile and in vivo pharmacokinetic (PK) parameters Following the outline of the procedure in Example 1, (2-hydroxypropyl)-β-cyclodextrin (HP-β-CD) or randomly methylated β-cyclodextrin (RM) Prepare an iloprostriposome composition containing (β-CD) and evaluate the IVR profile. did.

[0098] result: table 7 The following shows the physicochemical properties of iloprost liposome compositions containing different CDs. Both HP-β-CD and RM-β-CD reduced the burst release of the iloprost-liposome composition (less than 20% of iloprost was released within 1 hour of SLF incubation at 37°C).

[0099] [Table 7]

[0100] The above description includes many specific details to provide a complete understanding of the embodiments for illustrative purposes. However, it will be apparent to those skilled in the art that one or more other embodiments may be carried out without some of these specific details. Also, the terms “one embodiment,” “an embodiment,” and ordinal numbers throughout this specification refer to… References to embodiments should be understood to mean that certain features, structures, or characteristics may be included in the implementation of the disclosure. In the description, various features may be grouped together in a single embodiment, figure, or description thereof for the purpose of streamlining the disclosure and aiding in the understanding of various aspects of the invention, and one or more features or specific details of one embodiment may be implemented in conjunction with one or more features or specific details of other embodiments, if necessary, in the implementation of this disclosure. The present invention encompasses the following embodiments and forms. (1) A pharmaceutical composition comprising one or more liposomes suspended in an external medium, wherein the liposomes are (a) an outer lipid bilayer comprising at least one vesicle-forming phospholipid and less than 15 mol% sterols, and (b) An internal aqueous medium containing a weakly acidic drug and a weakly acidic salt. Includes, A pharmaceutical composition in which less than 65% of the weakly acidic drug is released into the external medium within one hour after administration of the pharmaceutical composition. (2) The pharmaceutical composition according to (1) above, wherein the outer lipid bilayer contains less than 10 mol% of sterols. (3) The pharmaceutical composition according to (1) above, wherein the outer lipid bilayer is substantially free of sterols. (4) The pharmaceutical composition according to (1) above, wherein the sterol is selected from the group consisting of cholesterol, cholesterol hexasuccinate, ergosterol, lanosterol, and combinations thereof. (5) The pharmaceutical composition according to (1) above, wherein the vesicle-forming phospholipid is a mixture of the first phospholipid and the second phospholipid or a mixture of the first phospholipid and a charged lipid. (6) The pharmaceutical composition according to (1) above, wherein the weakly acidic salt is a carboxylate or a bicarbonate. (7) The pharmaceutical composition according to (6) above, wherein the carboxylate salt is selected from the group consisting of formate, acetate, propionate, butyrate, isobutyrate, valerate, isovalerate, benzoate, and combinations thereof. (8) The pharmaceutical composition according to (6) above, wherein the bicarbonate is selected from the group consisting of potassium bicarbonate, sodium bicarbonate, calcium bicarbonate, magnesium bicarbonate, cesium bicarbonate, lithium bicarbonate, nickel bicarbonate, ferrous bicarbonate, or a combination thereof. (9) The pharmaceutical composition according to (1) above, wherein the internal aqueous medium further comprises cyclodextrin. (10) The pharmaceutical composition according to (9) above, wherein the molar ratio of the weakly acidic drug to the cyclodextrin (drug / CD ratio) is 0.06 or less. (11) The pharmaceutical composition according to (9) above, wherein the molar ratio of the weakly acidic drug to the cyclodextrin (drug / CD ratio) is 0.03 or less. (12) The pharmaceutical composition according to (1) above, wherein the weakly acidic drug is a prostaglandin, a prostacyclin receptor agonist, a steroid, a nonsteroidal anti-inflammatory drug (NSAID), an anticoagulant, an endothelin (ET) receptor antagonist, or a combination thereof. (13) The pharmaceutical composition according to (12) above, wherein the prostaglandin is iloprost. (14) The pharmaceutical composition according to (12) above, wherein the ET receptor antagonist is ambrisentan. (15) A method for treating a respiratory disease, comprising the step of administering the pharmaceutical composition described in (1) above. (16) A method for reducing side effects of a weakly acidic drug, comprising the step of administering an effective amount of the pharmaceutical composition described in (1) above to a subject that requires it. (17) The method according to (16) above, wherein the side effects of the weak acid drug in the upper respiratory tract are reduced by inhaling the weak acid.

Claims

1. A pharmaceutical composition comprising one or more liposomes suspended in an external medium, wherein the liposomes are (a) an outer lipid bilayer comprising at least one vesicle-forming phospholipid and less than 15 mol% sterols, and (b) An internal aqueous medium comprising (i) a weakly acidic drug which is treprostinil, iloprost, MRE-269, ambrisentan, bosentan, ketorolac, piroxicam, meloxicam, methylprednisolone, or dexamethasone, and (ii) a weakly acidic salt. Includes, A pharmaceutical composition wherein the pharmaceutical composition is incubated with simulated lung fluid (SLF), and less than 65% of the weakly acidic drug is released into the simulated lung fluid within one hour of incubation.

2. The pharmaceutical composition according to claim 1, wherein the external medium is a citrate buffer or a phosphate buffer.

3. The pharmaceutical composition according to claim 1, wherein the external lipid bilayer contains less than 13 mol% of sterols.

4. The pharmaceutical composition according to claim 1, wherein the external lipid bilayer contains less than 10 mol% of sterols.

5. The pharmaceutical composition according to claim 1, wherein the external lipid bilayer does not contain sterols.

6. The pharmaceutical composition according to claim 1, wherein the sterol is cholesterol, cholesterol hexasuccinate, ergosterol, lanosterol, or any combination thereof.

7. The pharmaceutical composition according to claim 1, wherein the vesicle-forming phospholipid is a mixture of a first phospholipid and a second phospholipid or a mixture of a first phospholipid and a charged lipid.

8. The pharmaceutical composition according to claim 7, wherein the first phospholipid is phosphatidylcholine (PC), and the second phospholipid is a polyethylene glycol-modified phospholipid or a negatively charged phospholipid.

9. The phosphatidylcholine (PC) is selected from the group consisting of hydrogenated egg phosphatidylcholine (HEPC), hydrogenated soybean phosphatidylcholine (HSPC), dipalmitoyl phosphatidylcholine (DPPC), distearoyl phosphatidylcholine (DSPC), diarachidoyl phosphatidylcholine, dimyristoyl phosphatidylcholine (DMPC), egg phosphatidylcholine (EPC), soybean phosphatidylcholine (SPC), oleoyl palmitoyl phosphatidylcholine, dioleoyl phosphatidylcholine (DOPC), dipetrocerinoyl phosphatidylcholine, palmitoyleleidoyl phosphatidylcholine, palmitoyloleoyl phosphatidylcholine, dilauroyl phosphatidylcholine (DLPC), diundecanoyl phosphatidylcholine, didecanoyl phosphatidylcholine, dinonanoyl phosphatidylcholine, and any combination thereof. The polyethylene glycol-modified phospholipid is 1,2-distearo-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG2000), The pharmaceutical composition according to claim 8, wherein the negatively charged phospholipid is distearoyl phosphatidylglycerol (DSPG), dipalmitoyl phosphatidylglycerol (DPPG), dimyristoyl phosphatidylglycerol (DMPG), or dioleoyl phosphatidylglycerol (DOPG).

10. The pharmaceutical composition according to claim 7, wherein the first phospholipid is phosphatidylcholine (PC), and the charged lipid is stearylamine, 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), 3β-[N-(N,N-dimethylaminoethane)-carbamoyl]cholesterol (DC-cholesterol), N4-cholesteryl-spermine (GL67), dimethyldioctadecylammonium (DDAB), 1,2-di-O-octadecenyl-3-trimethylammoniumpropane (DOTMA), ethylphosphocholine (ethylPC), or a combination thereof.

11. The pharmaceutical composition according to claim 7, wherein the first phospholipid is HSPC and the second phospholipid is DSPE-PEG2000 or DSPG.

12. The pharmaceutical composition according to claim 1, wherein the weakly acidic salt is a carboxylate or a bicarbonate.

13. The pharmaceutical composition according to claim 12, wherein the carboxylate salt is formate, acetate, propionate, butyrate, isobutyrate, valerate, isovalerate, benzoate, or any combination thereof.

14. The pharmaceutical composition according to claim 12, wherein the bicarbonate is potassium bicarbonate, sodium bicarbonate, calcium bicarbonate, magnesium bicarbonate, cesium bicarbonate, lithium bicarbonate, nickel bicarbonate, ferrous bicarbonate, or any combination thereof.

15. The pharmaceutical composition according to claim 1, wherein the internal aqueous medium further comprises cyclodextrin.

16. The pharmaceutical composition according to claim 15, wherein the molar ratio of the weakly acidic drug to the cyclodextrin (drug / CD ratio) is 0.06 or less.

17. The pharmaceutical composition according to claim 15, wherein the molar ratio of the weakly acidic drug to the cyclodextrin (drug / CD ratio) is 0.03 or less.

18. A pharmaceutical composition according to any one of claims 1 to 17 for the treatment of respiratory diseases.

19. The pharmaceutical composition according to claim 18, wherein the respiratory disease is pulmonary hypertension or interstitial lung disease.

20. A pharmaceutical composition according to any one of claims 1 to 17 for reducing the side effects of a weakly acidic drug, wherein the weakly acidic drug is inhaled.

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