OA20568A - Micromolecule PI4Klllα inhibitor composition, preparation method therefor and use thereof. - Google Patents

Micromolecule PI4Klllα inhibitor composition, preparation method therefor and use thereof. Download PDF

Info

Publication number
OA20568A
OA20568A OA1202200005 OA20568A OA 20568 A OA20568 A OA 20568A OA 1202200005 OA1202200005 OA 1202200005 OA 20568 A OA20568 A OA 20568A
Authority
OA
OAPI
Prior art keywords
pharmaceutical composition
composition according
pao
oil
lipid
Prior art date
Application number
OA1202200005
Inventor
Fude HUANG
Feng Wang
Shu Yang
Changping JIAO
Xiaojun Zhou
Original Assignee
Nuo-Beta Pharmaceutical Technology (Shanghai) Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nuo-Beta Pharmaceutical Technology (Shanghai) Co., Ltd. filed Critical Nuo-Beta Pharmaceutical Technology (Shanghai) Co., Ltd.
Publication of OA20568A publication Critical patent/OA20568A/en

Links

Abstract

Disclosed is a pharmaceutical composition comprising a micromolecule PI4Klllα inhibitor and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier comprises a lipid. Also disclosed are a method for preparing the pharmaceutical composition, and a method for treating PI4Klllα-related diseases by using the pharmaceutical composition.

Description

MKROMOLECULE ΡΙ4ΚΠΙΑ INHIBITOR COMPOSITION, PREPARATION METHOD THEREFOR AND USE THEREOF
TECHNICAL FIELD
[0001] The présent invention relates to a pharmaceutical composition, in particular to a pharmaceutical composition comprising a therapeutically effective amount of a micromolecule ΡΙ4ΚΙΠα inhibitor and a pharmaceutically acceptable carrier. The présent invention further relates to a préparation method for the pharmaceutical composition and use thereof.
BACKGROUND
[0002] Phosphatidylinositol 4-kinase (PI4KIIIa) is a kinase capable of catalyzîng phosphorylation of a D4 position on a phosphatidyl inositol (PI) ring to produce 4phosphatidyl-inositide (PI4P). The P14P is then catalyzed by PIP5-K kinases to generate 4,5phosphatidyl-inosididediphosphate (PIP2), and the PIP2 is a direct catalytic substrate of a PI3K, can actîvate the activities of multiple downstream proteins and plays a key rôle in PI3K/Akt. Therefore, the ΡΙ4ΚΙΠα indirectly affects a PI3K/Akt signaling pathway by affecting the PIP2, and a ΡΙ4ΚΙΠα inhibitor can be thus used for treating diseases related to the PI3K/Akt signaling pathway.
[0003] Particularly, studîes hâve shown that the PI4P, a product of the PI4KIIIa, is significantly increased in the cérébral cortex of an Alzheimer's disease (AD) patient, and the increased level is closely related to the degree of cognitive dysfunction in the AD patient (Zhu, L., et al., Proc Natl Acad Sci USA, 2015). In AD models of cultured cells, drosophilae and mice, inhibiting the ΡΙ4ΚΠΙα through genetic methods or compounds can promote the release of β-amyloid peptide 42 (Αβ42) from cells and relieve neurological damage on the AD animal models, including synaptic transmission as well as leaming and memory disorders (Zhang, X,, et al, J. Neurosci, 2017; Zhang et al., 2017;Huang. FD., et al·, PCT/CN2016/080907). Therefore, the ΡΙ4ΚΠΙα kinase inhibitor can effectively treat the AD. [0004] The ΡΙ4ΚΠΙα inhibitor may bave many therapeutic uses, but such inhibitor has the disadvantages such as low water solubility and poor stability. The PI4KIIIa inhibitor may be delivered by organic solvents commonly used for such médicament or other methods that promote the solubilization of such médicament in water, but the use of such préparations to deliver the PI4KIIIa inhibitor in vivo leads to poor bioavailability, it is impossible to avoid or reduce the toxîcîty of the médicament itself in the body (e.g., in the digestive tract), and the organic solvents themselves also hâve a risk of potential toxîcîty. Therefore, there is currently
a need for a pharmaceutical préparation of the PI4KIIIa inhibitor that can be effectîvely delivered and minimize the toxicity of active substances.
SUMMARY
[0005] The présent disclosure provides a pharmaceutical composition comprising a 5 micromolecule PI4KIIIa inhibitor and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier includes a lipid.
[0006] In some embodiments, the micromolecule PI4KIIIa inhibitor is PAO and a dérivative of PAO.
[0007] In some embodiments, the micromolecule PI4KIIIa inhibitor has a structure of 10 fonnula (I) or a pharmaceutically acceptable sait thereof.
formula (I) wherein Ri is each independently selected from H, halogen, nitro, cyano, hydroxyl, amino, carbamoyl, Ci-e alkyl, Ci-e alkoxy, Ci-6 haloalkyl, -As(O), -NH-(Cj-6 alkyl), N,N-(Ci-6 alkyl)2, 15 -NH-C(O)-R2, -NH-S(O)2-Rs, -C(O)ORa or heterocyclyl, wherein n is an mteger of 0-5, R:
and Ri are each independently selected from H, amino, Ci-6 alkyl, Ci-6 alkoxy. Ci.6 haloalkyl, -NH-(Ci-e alkyl), N,N-(Ci-e alkyl)2, -C(O)OR4, C3.6 cycloalkyl, 6-12 membered aryl or 3-6 membered heterocyclyl, which are optionally substituted by halogen, nitro, cyano, hydroxyl, amino, carbamoyl, aryl, Ci-6 alkyl, C2^ alkynyl, C2-6 alkenyl, Ci-6 alkoxy, Ci-e haloalkyl, 3-6 20 membered heterocyclyl, C3-6 cycloalkyl or Bn-O-, and R4 is Cj-6 alkyl.
[0008] In some embodiments, Ri is each independently selected from H, halogen, nitro, cyano, hydroxyl, amino, carbamoyl, Ci-e alkyl, Ci-e alkoxy, Ci-6 haloalkyl, -As(O), NH-(Ci-ô alkyl), N,N-(Cj-6 alkylfr or -C(O)OR4, wherein n is an integer of 0-2, and R4 is C1-6 alkyl. In some embodiments, Ri is each independently selected from H, halogen, nitro, cyano, 25 hydroxyl, amino, Ci-6 alkyl, C1-6 alkoxy. Ci-6 haloalkyl or -As(O), wherein n is an integer of
0-2. In some embodiments, Ri îs each independently selected from H, halogen, amino or Ci-e alkoxy, wherein n is 1. In some embodiments, Ri is located at an ortho position or a para position of the -As(O) group. In some embodiments, Ri is H.
[0009] In some embodiments, the micromolecule PI4KIIIa inhibitor is at an amount 30 of 0.01-20 mg/g, 0.05-20 mg/g, 0.1-20 mg/g, 0.2-20 mg/g, 0.5-20 mg/g, 0.8-20 mg/g, 1-20 mg/g, 1-18 mg/g, 1-16 mg/g, 1-14 mg/g, 1-12 mg'g, 1-10 mg/g, 2-10 mg/g, 2-8 mg/g, 2-6 mg/g, 3-6 mg/g 0.2-15 mg'g, 0.2-12 mg'g 0.2-10 mg/g, 0.2-8 mg'g, 0.2-6 mg/g, 0.2-4 mg'g, 0.2-2 mg/g, 0.2-1 mg'g or 0.2-0.8 mg/g in the pharmaceutical composition.
[0010] In some embodiments, the pharmaceutically acceptable carrier comprises at least about 50% (w/w), at least about 60% (w/w), at least about 70% (w/w), at least about 80% (w/w), at least about 85% (w/w), at least about 90% (w/w), at least about 95% (w/w), at least about 97% (w/w), at least about 98% (w/w), at least about 99% (w/w) or 100% (w/w) ofthe lipid.
[0011] In some embodiments, the lipid comprises a lipid with a melting point of -2080°C,-20-10°C or-20-0°C.
[0012] In some embodiments, the lipid has a degree of unsaturation of 0-5, 0-4, 0-3, 0-2, 0-1 orO.
[0013] In some embodiments, the lipid comprises a lipid which has a fatty acid carbon chain at a length în a range of 4-24, 4-22, 4-20, 6-20, 6-16, 6-14, 6-13, 6-12, 8-13, 812 or 8-10 carbon atoms.
10014] In some embodiments, the lipid comprises a lipid which has a fatty acid chain at a length of 8 and 10, and optionally further comprises a lipid which has the fatty acid carbon cham at a length of 12-22.
[0015] In some embodiments, the fatty acid chain in the lipid is a long-chain fatty acid, a medium-chain fatty acid or a short-chain fatty acid.
[0016] In some embodiments, the lipid is vegetable oil. In some embodiments, the vegetable oil is olive oil, tea oil, rapeseed oil, peanuî oil, soybean oil, corn oil, safflower oil, groundnut oil, sunflower seed oil, canola oil, walnut oil, almond oil, avocado oil, castor oil, coconut oil, cottonseed oil, rice bran oil, sesame oil, refrned palm oil or a mixture thereof.
[0017] In some embodiments, the lipid is a fatty acid, a fatty acid ester, a fatty alcohol, a lipoid, a paraffin or a mixture thereof.
[0018] In some embodiments, the lipoid is a phospholipid, a sucrose ester, a steroid, a fat-soluble vitamin or a mixture thereof.
[0019] In some embodiments, the fatty acid ester is a glyceride, an ethylene glycol ester, a propylene glycol ester or a mixture thereof. In some embodiments, the fatty acid ester is a monoester, a dîester, a triester or a mixture thereof. In some embodiments, the fatty acid ester comprises glycerides of octanoic acid and/or decanoic acid. In some embodiments, the fatty acid ester is substantially consisting of glycerides of octanoic acid and/or decanoic acid.
In some embodiments, the fatty acid ester comprises a medium-chain triglycéride. In some embodiments, the fatty' acid ester is a medium-chain triglycéride.
[0020] In some embodiments, the pharmaceutically acceptable carrier does not comprise an unsaturated lipid.
[0021] In some embodiments, the pharmaceutically acceptable carrier further comprises an antioxidant. In some embodiments, the antîoxidant is at an amount of 0.001%-5% (wt), 0.005%-5% (wt), 0.01%-5% (wt), 0.05%-5% (wt), O.l%-5% (wt), 0. l%-3% (wt), 0.1%2% (wt), 0.1%-l% (wt), 0.1%-0.8% (wt), 0.1%-0.5% (wt), 0.1%-0.3% (wt), 0.3%-2% (wt), 0.5%-2% (wt), O.8%-2% (wt) or l%-2% (wt) based on the weight of the pharmaceutical composition. In some embodiments, the antioxidant is sulfite, bisulfite, pyrosulfite, dithiocarbamate, ascorbic acid, ascorbyl palmitate, hydrocoumarin, vilamin E. ethanolamine, propyl gallate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), nordihydroguaiaretic acid or glutathione.
[0022] In some embodiments, the pharmaceutically acceptable carrier does not comprise an antioxidant.
[0023[ In some embodiments, the pharmaceutically acceptable carrier further comprises a viscosity modifier, a pH regulator or a flavoring agent.
[0024] In some embodiments, the pharmaceutically acceptable carrier further comprises ethanoL In some embodiments, the éthanol is at an amount of 10%-0.1% (v/v). In some embodiments, the éthanol is at an amount of 8%-0.1% (v/v), 7%-0.1% (v/v), 6%-0.l% (v/v), 5%-0.1% (v/v), 4%-0.1% (v/v), 3%-0.1% (v/v), 2%-0.1% (v/v), 1.5%-0.1% (v/v), L2%-0.1% (v/v), 8%-O.3% (v/v), 8%-O.5% (v/v), 8%-0.7% (v/v), 8%-0.9% (v/v), 8%-l% (v/v), 6%-0.3% (v/v), 5%-0.5% (v/v), 4%-0.8% (v/v), 3%-0.9% (v/v) or 2%-l% (v/v).
[0025] In some embodiments, the pharmaceutical composition is used for oral, subcutaneous, intramuscular or intravenous administration.
[0026] In some embodiments, the pharmaceutical composition is tablets, capsules, suspensions, solutions, semisoîid préparations, patches or microneedles.
[0027] In some embodiments, the micro molécule PI4KIIIa inhibitor is phenylarsine oxide, the phenylarsine oxide is at an amount of 0.1 -20 mg/g in the pharmaceutical composition, and the pharmaceutically acceptable carrier is consisting of a medium-chain triglycéride, consisting of a medium-chain triglycéride and a long-chain triglycéride, or consisting of a medium-chain triglycéride and ethanoL
[0028] In some embodiments, phenylarsonic acid is at an amount of less than 5%, 4%, 3%, 2%, 1 %, 0.7%, 0.5%, 0.3% or 0.2% in the pharmaceutical composition. In some embodiments, the phenylarsonic acid is at an amount of less than 5%. 4%, 3%, 2%, 1%, 0.7%, 0.5%, 0.3% or 0.2% after the pharmaceutical composition îs stored under conditions of 25°C/60%RH for I month, 2 months, 3 months, 4 months, 5 months. 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 1.5 years, 2 years or 3 years. In some embodiments, the phenylarsonic acid is at an amount of less than 5%, 4%, 3%, 2%, 1%, 0,7%, 0.5%, 0.3% or 0.2% after the pharmaceutical composition is stored under a condition of 28°C for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 1.5 years, 2 years or 3 years.
(0029] In another aspect, the présent disclosure provides a method for preparing the pharmaceutical composition provided herein, The method comprises: mixing the mîcromolecule PI4KIIIa inhibitor and the pharmaceutically acceptable carrier to obtain a mixture.
[0030] In some embodiments, the method comprises: mixing the mîcromolecule ΡΙ4ΚΙΠα inhibitor and the pharmaceutically acceptable carrier through a niechanical force. In some embodiments, the mechanical force is stirrîng, dispersing, shakîng or ultrasonic treatment.
[0031] In some embodiments, the method comprises: mixing the mîcromolecule ΡΙ4ΚΙΠα inhibitor and the pharmaceutically acceptable carrier after melting the pharmaceutically acceptable carrier by heating.
[0032] In some embodiments, the method further comprises: filterîng the mixture.
[0033] In another aspect, the présent disclosure provides a method for treating a PI4KIIIa-related disease in a subject. The method comprises administrât!ng the pharmaceutical composition provided herein to a subject in need thereof.
[0034] In some embodiments, the PI4KIIIa-related disease is Alzheimer’s disease.
10035] In some embodiments, the subject is an animal such as a pig, a dog, a monkey, a cat, a mouse, or a rat, or a human.
[0036] In another aspect, the présent disclosure provides use of the pharmaceutical composition provided herein in the manufacture of a médicament for treating a PI4KIIIareiated disease in a subject.
[0037] In still another aspect, the présent disclosure provides the pharmaceutical composition provided herein for use in treating a P14KIIIa-related disease in a subject.
BRŒF DESCRIPTION OF THE DRAWINGS
[0038] FIG. 1 shows the dissolution profiles of the PAO. In the figure, the cumulative dissolution % of a sample at 60 min is shown as zéro because of data missing, not indicating that the cumulative dissolution % is zéro indeed.
[0039] FIG. 2 shows the in vitro release profiles of MCT solution samples.
[0040] FIG. 3 shows the in vitro release profiles of glyceryl behenate solid dispersion samples.
[0041] FIG. 4 shows the in vitro release profiles of MC suspensions.
[0042] FIG. 5 shows the in vitro release profiles of MC+0.1% Tween 80 suspensions.
[0043] FIG. 6A shows the blood concentrations of the PAO after intravenous administration ofthe PAO at 0.1 mg/kg; FIG. 6B shows the blood concentrations ofthe PAO after oral administration of the PAO at 0.2 mg'kg; and FIG. 6C shows the average blood concentrations of the PAO after intravenous or oral administration.
[0044] FIG. 7A shows the blood concentrations of the PAO after intravenous administration of the PAO at 0.1 mg/kg; FIG. 7B shows the blood concentrations of the PAO after oral administration of the PAO at 0.2 mg/kg; and FIG. 7C shows the average blood concentrations ofthe PAO after intravenous or oral administration.
[0045] FIG. SA shows the blood concentrations of the PAO after oral administration of the PAO in a DMSO solution at 0.1 mg/kg; and FIG. 8B shows the blood concentrations of the PAO after oral administration of the PAO in an MCT solution at 0.1 mg/kg.
[0046] FIG. 9 shows the weight changes of female mice 1.5 months after intragastric administration of the PAO in a 0.1% DMSO solution or the PAO in an MCT solution at 1.5 mgZkg/day, where and ** represent p value of less than 0.05 and 0.01, respectively.
DETAILED DESCRIPTION
[0047] The following description of the disclosure is merely intended to il lu strate various embodiments of the disclosure. The spécifie embodiments discussed are not to be construed as limitations on the scope of the disclosure. It will be apparent to one skilled in the art that various équivalents, changes, and modifications may be made without departing from the spirit and essence of the disclosure, and it is understood that such équivalent embodiments are to be included herein. Ail references cited herein, including publications, patents and patent applications are incorporated herein by reference in their entirety.
[0048] In an aspect of the présent disclosure, provided is a pharmaceutical composition comprising a micromoiecule PMKIIIa inhibitor and a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier comprises a lîpîd.
[0049| Micromoiecule ΡΙ4ΚΙΠα inhibitor |0050] As used herein, the term micromoiecule PMKIIIa inhibitor refers to various micromoiecule compounds that can reduce, decrease. or eliminate the transcription or translation of a PMKIIIa gene, and'or the concentration or activity of a PMKIIIa protein. In some embodiments, the micromoiecule PMKIIIa inhibitor is capable ofreducing the activity of the PMKIIIa by at least 5%, 10%, 20%, 40%, 50%, 80%, 90%, 95% or more.
[0051] In some embodiments, the micromoiecule PMKIIIa inhibitor is a micromoiecule organic or inorganic compound (e.g., a molécule obtained from an artificially synthesized Chemical library and a natural product library). In some embodiments, the micromoiecule PMKIIIa inhibitor has a moiecular weight of less than 3,000, 2,500, 2,000, 1,500, 1,000, 900, 800, 700, 600, 500, 400, 300, 250, or 200 Daltons.
[0052] In some embodiments, the micromoiecule PMKIIIa inhibitor directly binds to the PMKIIIa protein. In some embodiments, the micromoiecule PMKIIIa inhibitor specifically binds to the PMKIIIa protein.
[0053] As used herein, the term spécifie binding, when used to describe the PMKIIIa inhibitor, means that the PMKIIIa inhibitor preferably recognizes the PMKIIIa protein în a complex mixture, and the binding constant of the inhibitor to the PMKIIIa protein is at least 2 times as high as that of the inhibitor to other non-specific binding proteins. In certain embodiments, the equilibrium dissociation constant of the PMKIIIa inhibitor from the PMKIIIa protein is less than or equal to 10-î or ΙΟ’6 M. In certain embodiments, the equilibrium dissociation constant of the PMKIIIa inhibitor from the PMKIIIa protein is less than or equal to 10'6 or I0'7 M. In certain embodiments, the equilibrium dissociation constant of the PMKIIIa inhibitor from the PMKIIIa protein is less than or equal to 10'7 or 10 s M. [0054] In some embodiments, the micromoiecule PMKIIIa inhibitor provided herein is PAO and a dérivative of PAO.
[0055J As used herein, the term PAO refers to a micromoiecule compound with an arsenic oxide group and a benzene ring as basic structures. Its spécifie Chemical structure is as follows:
[0056] In the présent disclosure, the PAO and PI01 are used interchangeably. [0057] As used herein, the term a dérivative of PAO refers to a class of micromoiecule compounds dem ed from the PAO. These micromoiecule compounds hâve the same basic structures as the PAO (i.e., having an arsenic oxide group and a benzene ring), and can ail inhibit PI4KIIIa. In some embodiments, the inhibitory activity of the dérivative of PAO on ΡΙ4Κ1ΙΙα is at least 50%, 80%, 90%, 95%, 100%, 120%, 150%, I time, 2 times, 3 times, 4 times or more times as high as the inhibitory activity of the PAO. In some embodiments, the solubility of the dérivative of PAO in water is 50%-200%, 80%-180%, 90%-150%, 95%-150%, 100-150%, I2O%-15O%, 80%-150%, 80%-130%, 80%-120% or 90%-l 10% of the solubility of the PAO in water. In some embodiments, the solubility of the dérivative of PAO in the pharmaceutically acceptable carrier provided herein is 50%-200%, 80%-180%, 90%-150%, 95%-150%, 100-150%, 120%-150%, 80%-150%, 80%-I30%, 80%120% or 90%-] J 0% of the solubility of the PAO in the pharmaceutically acceptable carrier provided herein.
[0058] In some embodiments, the micromoiecule ΡΙ4ΚΙΠα inhibitor provided herein has a structure of formula (I) or a pharmaceutically acceptable sait thereof,
formula (I) wherein Ri is each independently selected from H, halogen, nitro, cyano, hydroxyl, amino, carbamoyl, Ci-è alkyl, Ci-û alkoxy, Ci-6 haloalkyl, -As(O), -NH-(Ci-û alkyl), N,N-(Ci-6 alkyl)2, -NH-C(O)-R2, -NH-S(O)2-Rs, -C(0)0R4 or heterocyclyl, wherein n is an integer of 0-5, R2 and R3 are each independently selected from H, amino, Cne alkyl, Ci-6 alkoxy, C1-6 haloalkyl, -NH-(Ci-e alkyl), N,N-(Ci-6 alkyl)2, -C(O)OR4, Cj.e cycloalkyl, 6-12 membered aryl or 3-6 membered heterocyclyl, which are optionally substîtuted by halogen, nitro, cyano, hydroxyl, amino, carbamoyl, aryl, Ci^ alkyl, C;^ alkynyk C?^ alkenyl, Ci-c alkoxy, Ci^ haloalkyl, 3-6 membered heterocyclyl, Cs-e cycloalkyl or Bn-O-, and Rj is Ci-6 alkyL
[0059] In some embodiments, n is 0,1,2 or 3. In some embodiments, n is 0, 1 or 2. In some embodiments, n is 0 or 1,
[0060] In some embodiments, Ri is each independently selected from H, halogen, nitro, cyano, hydroxyl, amino, carbamoyl, Ci-6 alkyl, C^ alkoxy, C1j6 haloalkyl. -As(O), NH-(Ci^ alkyl), N,N-(Ci^ alkyl); or -C(O)OR4, where n is an integer of 0-2, and R4 îs C1-6 alkyl.
[0061] In some embodiments, Ri is each independently selected from H, halogen, nitro, cyano, hydroxyl, amino, Ci^ alkyl, C^ alkoxy, Ci-6 haloalkyl or -As(O), where n is an integer of 0-2.
[0062] In some embodiments, Ri is each independently selected from H, halogen, amino or Cι-e alkoxy, where n is 1.
[0063] In some embodiments, Ri is located at an ortho position or a para position of the -As(O) group. In some embodiments, Ri is H.
[0064] As used herein, the term substituted, when referring to a Chemical group, means that one or more hydrogen atoms of the Chemical group are removed and substituted by a substituent.
[0065] As used herein, the term substituent has the common meaning well known in the art and refers to a Chemical moiety that is covalently attached to or fused to a parent group where appropriate.
[0066] As used herein, the term Cn-Cm” represents a range of the number of carbon atoms, where n and m are întegers, and the range of the number of carbon atoms includes endpoints (i.e., n and m) and every integer point therebetween. For example, Ci-Ce represents a range of 1 to 6 carbon atoms, including 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms and 6 carbon atoms.
[0067] As used herein, the tenu alkyl, whether used as part of other terms or used alone, refers to a saturated hydrocarbyl group, which may be linear or bran ch ed. The terni Cn-Cm alkyl refers to an alkyl having n to m carbon atoms. In certain embodiments, the alkyl group includes 1 to 12, 1 to 8, 1 to 6, 1 to 4, 1 to 3, or 1 to 2 carbon atoms. An example of the alkyl group includes, but is not limited to, a Chemical group such as methyl, ethyl, npropyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, 2-methyl-l-butyl, n-pentyl, 3-pentyI, n-hexyl, 1,2,2-trimethylpropyl, etc.
[0068] As used herein. the tenu alkenyl, whether used as part of other tenus or used alone, refers to an unsaturated hydrocarbyl group, which may be linear or branched and has at least one carbon-carbon double bond. In certain embodiments, the alkenyl group includes 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 5, 2 to 4, or 2 to 3 carbon atoms. In certain embodiments, the alkenyl group can also hâve 1 to 6, 1 to 5, 1 to 4, I to 3, 1 to 2, or 1 carbon-carbon double bond. An example of the alkenyl group includes, but is not limited to, a Chemical group such as vinyl, n-propenyl, isopropenyl, π-butenyl, sec-butenyl, etc.
[0069] As used herein, the term alkynyl, whether used as part of other terms or used alone, refers to an unsaturated alkynyl group, which may be linear or branched and lias ai least one carbon-carbon triple bond. In certain embodiments, the alkynyl group includes 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 5, 2 to 4, or 2 to 3 carbon atoms. In certain embodiments, the alkynyl group can also hâve 1 to 6, 1 to 5, 1 to 4, I to 3, I to 2, or 1 carbon-carbon triple bond. An example of the alkynyl group includes, but is not limited to, a Chemical group such as ethynyL propynyl, butynyl, etc.
[0070] As used herein, the term cycloalkyl refers to a cycîic alkyl consisting of at least 3 atoms. The term n-m membered cycloalkyl refers to a cycloalkyl having n to m ringforming members. In addition, the ring may also hâve one or more double bonds, but not a fuilly conjugated System. In certain embodiments, the cycloalkyl has 3 to 8, 3 to 6, or 4 to 6 ring-forming carbon atoms. An example ofthe cycloalkyl includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, etc.
[0071] As used herein, the term heterocyclyl refers to a cyclyl of which at least one atom in the ring system is a heteroatom and the remaining ring atoms are carbon atoms. The term n-m membered heterocyclyl refers to a heterocyclyl having n to ni ring-forming members. As used herein, the term heterocyclyl includes heteroaryl and heterocycloalkyl. In addition, the ring may also hâve one or more double bonds. In certain embodiments, the heterocyclyl is a saturated heterocycloalkyl. An example ofthe heteroatom includes, but is not limited to, oxygen, sulfur, nitrogen, phosphorus, etc.
[0072) As used herein, the term heterocycloalkyl refers to a cycloalkyl of which at least one atom in the ring system is a heteroatom and the remaining ring atoms are carbon atoms. The term n-m membered heterocycloalkyl refers to a heterocycloalkyl having n to m ring-forming members. In addition, the ring may also hâve one or more double bonds, but not a fully conjugated system. In certain embodiments, the heterocycloalkyl is a saturated heterocycloalkyl. An example of the heteroatom includes, but îs not limited to, oxygen, sulfur, nitrogen, phosphorus, etc. In certain embodiments, the heterocycloalkyl has 3 to 8, 3 to 6, or to 6 ring-forming carbon atoms. An example of the heterocycloalkyl includes, but is not limited to, azetidîne. aziridine, pyrrolidinyl, piperîdinyl, piperazinyl, morpholinyl. thiomorpholine, homopiperazine, etc.
[0073] As used herein, the term aryl or ”aromatic group, whether used as part of other ternis or used alone, refers to a single-carbocycle or multi-carbocycle cyclic group having alternate double bonds and single bonds between ring-forming carbon atoms. The term Cn-Cni aryl refers to an aryl having n to m ring-forming carbon atoms. In certain embodiments, an aryl ring System has 6 to 12, 6 to 10, or 6 to 8 carbon atoms in one or more rings. In certain embodiments, the aryl ring System has 2 or more rings fused together. An example of the aryl group includes, but is not limited to, a Chemical group such as phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl, etc.
[0074] As used herein, the term heteroaryl refers to an aryl group of which at least one ring atom in the aromatîc ring is a heteroatom and the remaining ring atoms are carbon atoms. The term n-m membered heteroaiyl refers to a heteroaryl having n to ni nngforrning members. An example of the heteroatom includes, but is not limited to, oxygen, sulfur, nitrogen, phosphorus, etc. In certain embodiments, the heteroaryl may hâve 5 to 10, 5 to 8, or 5 to 6 ring-forming members. In certain embodiments, the heteroaryl is a 5 or 6 membered heteroaryl. An example of the heteroaryl includes, but is not limited to, furyl, thienyl, pyridyl, quinolinyl, pyrrolyl, N-lower alkylpyrrolyl, pyndyl-N-oxide, pyrimidinyl, pyrazinyl, imidazolyl, indolyl, etc.
[0075] As used herein, the term alkoxy, whether used as part of other terms or used alone, refers to a group of formula -O-alkyl. The tenu Cn-Cm alkoxy means that an alkyl moiety ofthe alkoxy has n to m carbon atoms. In certain embodiments, the alkyl moiety has 1 to 6, 1 to 4, or 1 to 3 carbon atoms. An example of the alkoxy group includes, but is not limited to, a Chemical group such as methoxy, ethoxy, propoxy (e.g., ίί-propoxy and îsopropoxy), ί-butoxy, etc.
[0076] As used herein, the term haloaikyl, whether used as part of other terms or used alone, refers to a group of formula -alkyi-X, where X is halogen, an atom selected from fluorine, chlorine, bromine and iodine. The term Cn-Cm haloaikyl means that an alkyl moiety of the haloaikyl has n to m carbon atoms. In certain embodiments, the alkyl moiety has 1 to 6, 1 to 4, or 1 to 3 carbon atoms. An example of the haloaikyl group includes, but is not limited to, a Chemical group such as halomethyl, haloethyl, halopropyl (e.g., n-halopropyl and isohalopropyl), Z-halobutyl, etc.
[0077] As used herein, the term n membered” is usually used with a ring System to describe the number of ring-forming atoms in the ring System, where n is an integer. For example, piperidinyl is an example of a 6 membered heterocycloalkyl ring, pyrazolyl is an example of a 5 membered heteroaryl ring, pyridyl is an example of a 6 membered heteroaryl ring, and 1,2,3,4-tetrahydro-naphthalene is an example of a 10 membered aryl.
[0078] As used herein, the term halogen refers to an atom selected from fluorine, chlorîne, bromîne and iodine.
[0079] As used herein, the term cyano refers to a group of fonnula -CN.
[0080] As used herein. the term hydroxyl refers to a group of formula ”-OH.
|0081] As used herein, the term nitro refers to a group of fonnula -NOz”.
[0082] As used herein, the term amino refers to a group of fonnula -NH2.
[0083] As used herein, the term carbamoyl refers to a group of formula
HNCONH2.
[0084] As used herein, the tenu compound is intended to include ail stereoisomers (e.g., enantiomers and diastereomers), géométrie isomers, tautomers and isotopes of the shown structure.
[0085] The compound provided herein may be asymmetric (e.g., having one or more stereocenters). Unless otherwise indicated, ail the stereoisomers, such as enantiomers and diastereomers, are intended to be included. The compound provided herein including asymmetricaliy substituted carbon atoms may be separated in an optical ly activated or racemic form. Methods to préparé the optically active form from starting materials that are not optically active are well known in the art, such as by resolution of racemic mixtures or by stereoselective sÿnthesis. Various géométrie isomers, such as olefins, carbon-carbon double bonds and the like, may also exist in the compound provided herein, and ail of these stable isomers hâve been considered in the present disclosure. The present disclosure describes cis and trans géométrie isomers of the compound, which may be separated as a mixture of isomers or as individual isomers.
[0086] In certain embodiments, the compound provided herein has a (Λ)configuration. In certain embodiments, the compound provided herein has a (^-configuration. [0087] The racemic mixture of the compound may be resolved by any one of multiple methods well known in the art. An exemplary method includes fractional crystailization using a chiral resolving acid which is an optically active salt-forming organic acid. Suitable resolving reagents for the fractional recrystallization method are, for example, optically active acids (e.g., D and L forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid,
mandelic acid, maiic acid, iactic acid) or varions optically active camphorsulfonic acids.
Other resolving reagents suitable for the fractional crystallization method include stereoisomerically pure forms of N-methylbenzylamine, 2-phenylgIycinol, norephedrine, ephedrine, Mmethylephedrine, cyclohexylethylamme, 1,2-diaminocyclohexane. etc.
[0088] The racemic mixture may also be resolved by elution on a column provided with an optically active resolving reagent (e.g., dînîtrobenzoylphenylglycine). A suitable elution solvent composition may be determined by a person skilied in the art.
[0089] The compound provided herein also includes tautomeric forms. The tautomeric fonns are caused by the interconversion between a single bond and an adjacent 10 double bond both accompanied by the migration of protons. The tautomeric forms include tautomers of protons in an isomeric protonated State with the same Chemical formula and total charge. Examples of the proton tautomers include a keto-enol pair, an amide-imidic acid pair, a lactam-lactim pair, an enamine-imine pair, and an annular form in which protons can occupy two or more positions of a heterocyclic system, such as 1H- and 3H-imidazole, 1H-, 15 2H- and 4H-l,2,4-triazole, ÎH- and 2H-isoindole, and 1H- and 2H-pyrazoie. The tautomeric forms can be balanced or stericaliy locked into one form through appropriate substitution.
[0090] The compound provided herein can also include ail isotopes of atoms existing in intermediate or final compounds. The isotopes include those atoms with the same atomic number but different mass numbers. For example, isotopes of hydrogen include protium, 20 deuterium and tritium.
[0091] In certain embodiments, the mîcromolecule compound provided herein may be obtained by organic synthesis. The compound provided herein, including salts, esters, hydrates or solvatés thereof, may be prepared by any well-known organic synthesis technology and may be synthesized according to many possible synthesis routes.
[0092] The reaction for preparing the compound provided herein may be carried out in a suitable solvent, and a person skilied in the field of organic synthesis can easily select the solvent. The suitable solvent cannot substantially react with starting materîals (reactants), intennediates or products at the reaction température (for example, the température may range from a freezing température of the solvent to a boiling température of the solvent). A given reaction may be carried out in one solvent or a mixture of more than one solvent.
According to spécifie reaction steps, a person skilied in the art can select suitable solvents for the spécifie reaction steps.
[0093] The préparation of the compound provided herein may involve the protection and deprotection of varions Chemical groups. A person skilied in the art can easily détermine whether protection and deprotection are needed and select suitable protective groups. Chemistry of the protective groups can be found m, for example. T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis. 3rd Ed., Wiley & Sons, Inc., New York (1999), the entrée contents of which are incorporated into the présent disclosure by reference. [0094] The reaction may be monitored according to any suitable method well known in the art. For example, the formation of products may be monitored by using spectroscopy, such as nuclear magnetic résonance spectroscopy (e.g., Ή or 13C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), mass spectrometry; or by using chromatography, such as high performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LCMS) or thin-layer chromatography (TLC). A person skilled în the art may purîfy the compound by many methods, including high performance liquid chromatography (HPLC) (see, for example, Préparative LC-MS Purification: Improved Compound Spécifie Method Optimization Karl F. Blom, Brian Glass, Richard Sparks, Andrew P. Combs J. Combi. Chem. 2004, 6(6), 874-883, the entrée contents of which are incorporated into the présent disclosure by reference) and normal-phase silica gel column chromatography.
|0095] In certain embodiments, the micromolecule compound provided herein may be commercially available.
[0096] In some embodiments, the micromolecule PMKIIIa inhibîtor provided herein îs at an amount of 0.01-20 mg/g, 0.05-20 mg/g, 0.1-20 mg/g, 0.2-20 mg/g, 0.5-20 mg/g, 0.820 mg/g, 1-20 mg/g, 1-18 mg/g, 1-16 mg/g, 1-14 mg/g, 1-12 mg/g, 1-10 mg/g, 2-10 mg/g, 2-8 mg/g, 2-6 mg/g, 2-5 mg/g, 2-4 mg/g, 2-3 mg/g, 3-6 mg/g, 0.2-15 mg/g, 0.2-12 mg/g. 0.2-10 mg/g, 0.2-8 mg/g, 0.2-6 mg/g, 0.2-4 mg/g, 0.2-2 mg/g, 0.2-1 mg/g or 0.2-0.8 mg/g in the pharmaceutical composition.
|0097] Pharmaceutically acceptable carrier
[0098] As used herein, the term pharmaceutically acceptable refers to those compounds, materials, compositions and/or dosage fonns that are suitable for use in contact with human and animal tissues within the scope of reasonable medical judgment without excessive toxicity, irritation, allergie reaction or other problems or complications, and hâve a reasonable benefit/risk ratio. In certain embodiments, the pharmaceutically acceptable compounds, materials, compositions and/or dosage forms refer to those used for animais (more particularly for humans) approved by regulatory authorities (e.g., U.S. Food and Drug Administration, State Food and Drug Administration or European Medicines Agency) or listed in widely accepted pharmacopoeia (e.g., U.S. Pharmacopoeia, Pharmacopoeia of the People's Republic of China or European Pharmacopoeia).
[0099] Pharmaceuticaliy acceptable carriers that may be used in the pharmaceutical composition provided herein include, but are not limited to, for example pharmaceuticaliy acceptable liquid, gel or solid vehicles, aqueous media (e.g,, sodium chloride injection, Ringer's solution injection, isotonie glucose injection, stérile water injection, or glucose and lactated Ringer's injection), non-aqueous media (e.g., plant-derived nonvolatile oil, cottonseed oil, corn oil. sesame oil, peanut oil or mediummedium-to-long-chain glyceride, such as medium-chain triglycéride), antimicrobial substances, isotonie substances (e.g., sodium chloride or glucose), buffers (e.g., phosphate or citrate buffers), antioxîdants (e.s., sodium bisulfate), anesthetics (e.g., procaine hydrochloride), suspending agents dîspersins agents (e.g., sodium carboxymethyl cellulose, hydroxypropyl methylcelhilose, or polywmylpynOlidone), chelating agents (e.g., EDTA (ethylenediaminetetraacetic acid) or EGTA (ethylene glycol bis(2-aminoethyl ether)tetraacetic acid)), emulsifiers (e.g., Polysorbate 80 (Tween-80)), diluents, adjuvants, or nontoxic auxiliary substances, other components well known in the art, or various combinations of the above. Suitable components may include, for example, fillers, binders, disintegrants, buffers, preservatives, lubrîcants, flavoring agents, thickeners, colorants or emulsifiers.
|0100] In some embodiments, the pharmaceuticaliy acceptable carrier provided herein further includes an antioxidant, such as sulfite, bisulfite, pyrosulfite, dithiocarbamate, ascorbic acid, ascorbyl palmitate, hydrocoumarin, vitamin E, ethanolamine, propyl gallate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), nordihydroguaiaretic acid or glutathione. In some embodiments, the antioxidant provided herein is at an amount of 0.001%-5% (wt), 0.005%-5% (wt), 0.01%-5% (wt), 0.05%-5% (wt), 0.1%-5% (wt), 0.1%-3% (wt), 0.1%-2% (wt), 0.1%-!% (wt), 0.1%-0.8% (wt), 0J%-0.5% (wt), 0.1%-0.3% (wt), 0.3%-2% (wt), 0.5%-2% (wt), 0.8%-2% (wt) or l%-2% (wt) based on the weight ofthe pharmaceutical composition.
[0101] In some embodiments, the pharmaceuticaliy acceptable carrier provided herein does not comprise antioxîdants.
[0102] In some embodiments, the pharmaceuticaliy acceptable carrier provided herein further comprises a viscosity modifier, a pH regulator or a flavoring agent.
[0103] The pharmaceutical composition provided herein may be used in administration routes well known in the art, such as injection administration (e.g., subeutaneous injection, intraperitoneal injection, intravenous injection (including intravenous drip or intravenous infusion), intramuscular injection or intradermal injection) or noninjection administration (e.g., oral administration, nasal administration, sublingual administration, rectal administration or external administration). In some embodiments, the pharmaceutical composition provided herein is used for oral, subcutaneous, intramuscular or intravenous administration.
[0104] In some embodiments, the pharmaceutical composition provided herein may be prepared into dosage fonns for oral administration (including but not limîted to capsules, tablets, pîlls, aqueous suspensions or solutions), dosage forms for injection administration (including but not limited to solutions, émulsions, liposomes, powder injections), suppositories for rectal administration, and dosage forms for topical administration (including but not limited to ointments, pastes, creams. lotions, gels, powder, solutions, sprays, inhalants or patches), etc.
|0105] In some embodiments, the pharmaceutical composition provided herein is tablets, capsules, suspensions, solutions, semisoiid préparations, patches or microneedles.
[0106] In some embodiments, the pharmaceutical composition provided herein is an oral liquid. As used herein, the term oral liquid is a liquid dosage form for oral administration, which includes (but is not limited to) pharmaceutically acceptable émulsions, microemulsions, solutions, suspensions, syrups and élixirs. In addition to active compounds, the liquid dosage form may include commonly used inert diluents (e.g., water or other solvents), solubilizers, emulsifiers, wetting agents, emulsifiers and suspending agents, sweetening agents, flavoring agents and fragrances. In some embodiments, the oral liquid is in the form of a solution. In some embodiments, the oral liquid may be diluted with a diluent before being administered to a patient. In some embodiments, the diluent is végétable oil, or an aqueous solution having a certain flavoring effect, such as soda water, fruit juice, etc.
[0107] In some embodiments, the pharmaceutical composition provided herein is an injection.
[0108] As used herein, the term injection refers to a préparation for injection, in which médicaments are formulated into solutions (aqueous or non-aqueous), suspensions or émulsions and filled into ampoules or multi-dose containers. The injection, such as a stenle injectable aqueous or oily suspension, may be formulated according to known technologies using suitable dispersing agents or wetting agents, suspending agents and emulsifiers. In some embodiments, the pharmaceutical composition provided herein is an oily injection. In some embodiments, the pharmaceutical composition provided herein is an injection including the lipid provided herein. In some embodiments, the pharmaceutical composition provided herein is an injection including mono-/di-glycerides of octanoic/decanoic acid or medium chain triglycérides. In some embodiments, the pharmaceutical composition provided herein is prepared into a pre-filled dosage form.
[0109] In some embodiments. the pharmaceutical composition provided herein is patches.
[0110] As used herein, the term patch refers to a flaky préparation which is made from active pharmaceutical ingrédients and suitable materials and may produce systemic or topical effects when pasted on the skin. The patch is consisting of a backing layer, a medicament-containing matrix, a pressure-sensitive adhesive and an anti-sticking layer to be removed before use. The patch may be used on intact skin surfaces, and may be also used on diseased or incomplète skin surfaces. The patch which is used on the intact skin surfaces and can diffuse médicaments through the skin into the blood circulation system is known as a transdermal patch. The action time of the transdermal patch is detennined by its médicament content and release rate. The patch may be classified into an adhesive dispersion type, a reservoir type and a peripheral adhesive type. In some embodiments, the pharmaceutical composition provided herein is a patch including the lipid provided herein. In sonie embodiments, the pharmaceutical composition provided herein is a patch including mono-/diglycerides of octanoic/decanoic acid or medium-chain triglycérides.
(0111J In some embodiments, the pharmaceutical composition provided herein is microneedles.
[0112] As used herein, the tenu microneedle refers to a préparation having a microneedle array that can pierce the stratum comeum to facilitate transdermal delivery of therapeutic agents. In sonie embodiments, the microneedle has a microneedle array with a height of 300 to 1,000 pm. The microneedle used herein may be made of a material including resin or other polymer materials, ceramies or metals. In addition, the material of the microneedle îs preferably a material including thermoplastic resin, and more preferably a material including biodégradable thermoplastic resin. In some embodiments, the pharmaceutical composition provided herein îs a microneedle including the lipid provided herein. In some embodiments, the pharmaceutical composition provided herein is a microneedle including niono-/di-glycerides of octanoic/decanoic acid or medium-chain triglycérides. In some embodiments, the pharmaceutical composition provided herein and the microneedle are prepared separately, but used in combination. In some embodiments, the pharmaceutical composition provided herein is used before or after the microneedle, for example, the microneedle is firstly applied to the skin of a patient, and then the pharmaceutical composition provided herein îs applied to the same site; altematively the
I pharmaceutical composition provided herein is firstly applied to the skîn of the patient, and then the mîcroneedie is applied to the same site.
[0113] Lioid
[0114] In some embodiments. the pharmaceutically acceptable carrier provided herein includes a iipid.
[0115] As used herein, the term Iipid refers to an ester and dérivatives thereof formed by the reaction of a fatty acid and an alcohol. It is a type of compounds general ly insoluble in water but soluble in fat-soluble solvents. It may be synthetic, semisynthetic or naturally occurring, including a fat, a phospholipid, a glycolipid, a cholestérol, a cholestérol ester, etc,
[0116] In some embodiments, the pharmaceutically acceptable carrier provided herein includes at least about 50% (w/w), at least about 60% (w/w), at least about 70% (w/w), at least about 80% (w/w), at least about 85% (w/w), at least about 90% (w/w), at least about 95% (w/w), at least about 97% (w/w), at least about 98% (w/w), at least about 99% (w/w) or 100% (w/w) of the Iipid.
[0117] In some embodiments, the Iipid provided herein includes a Iipid with a melting point of-20-80°C, -20-10°C or -20-0°C. In some embodiments, the lipîd provided herein includes a Iipid which is a liquid at room température. In some embodiments, the Iipid provided herein is consisting of a Iipid with a melting point of -20-0°C.
[0118] As used herein, the terni melting point refers to a température at which the solid state and the liquid state of a substance are in equilibrium under a certain pressure, that is, at this pressure and this melting point température, the Chemical potential of a substance in the solid state is equal to that in the liquid state, When the substance is pure, it generally has a ftxed melting point, that is, under a certain pressure, the température différence from initial melting to full melting (the range is known as a melting range) does not exceed 0.5-l°C. The melting point may be measured by conventional methods in the art, including but not limited to capillary measurement, microscope hot plate measurement, automatic melting point measurement, etc. In some embodiments, the melting point provided.herein is measured under normal pressure.
[0119] In some embodiments, the Iipid provided herein has a degree of unsaturation of 0-5, 0-4, 0-3, 0-2, 0-1 or 0. In some embodiments, the Iipid provided herein has a degree of unsaturation of 0 or 1. In some embodiments, the Iipid provided herein has a degree of unsaturation of 0.
[0120] As used herein, the term degree of unsaturation'’, also known as an index of hydrogen deficiency or a ring-plus-double-bond index, is a quantitative indicator of the degree of unsaturation of an organic molecuie, that is, for every 2 hydrogen atoms reduced in the organic molecuie as compared with an open-chain alkane with the same number of carbon atoms, the degree of unsaturation of the organic substance is increased by 1. In general, the degree of unsaturation is represented by a Greek letter Ω. The degree of unsaturation may help to détermine how many rings (1 degree of unsaturation), double bonds (1 degree of unsaturation) and triple bonds (2 degrees of unsaturation) a compound has. In some embodiments, the degree of unsaturation provided herein excludes the degree of unsaturation resulting from rings.
[0121] According to the degree of saturation, the lipid can be divided into two classes, namely a saturated lipid and an unsaturated lipid. According to the degree of unsaturation, the unsaturated lipid is further divided into a monounsaturated lipid and a polyunsaturated lipid. The monounsaturated lipid has only one double bond in the molecular structure; and a polyunsaturated fatty acid has two or more double bonds in the molecular structure.
[0122] In some embodiments, the pharmaceutically acceptable carrier provided herein dose not comprise unsaturated lipids.
[0123] In some embodiments, the lipid provided herein includes a lipid which has a fatty acid carbon chain at a length in a range of 4-24, 4-22, 4-20, 6-20, 6-16, 6-14, 6-13, 6-12, S-13, 8-12 or 8-10 carbon atoms. In some embodiments, the lipid provided herein includes a lipid which has a fatty acid chain at a length of 8 and 10, and optionally further includes a lipid which has the fatty acid carbon chain at a length of 12-22.
[0124] As used herein, the term fatty acid carbon chain length refers to the number of carbon atoms in a carbon chain in a fatty acid of the lipid.
[0125] In some embodiments, the fatty acid chain in the lipid is a long-chain fatty acid, a medium-chain fatty acid or a short-chain fatty acid. In some embodiments, the pharmaceutically acceptable carrier provided herein is consisting ofa medium-chain triglycéride, or consisting of a mixture of a medium-chain triglycéride and a long-chain triglycéride.
[0126] As used herein, the term long-chain fatty acid, also known as a higher fatty acid, refers to a fatty acid with more than 12 carbon atoms on a carbon chain. The long-chain fatty acid mainly exîsts in a natural fat and is a main component of the fat. There are many kinds of long-chain fatty acids in the natural fat. Common ones are palmitic acid (hexadecanoic acid), stearic acid (octadecanoic acid) and oleic acid (pctadecene-[9]-acid).
]0127] As used herein, the term medium-chain fatty acid refers to a fatty acid with 6-12 carbon atoms on a carbon chain, and main components are octanoic acid (C8) and decanoic acid (CIO).
[0128] As used herein, the term short-chain fatty acid, also known as a volatile fatty acid, refers to an organic fatty acid with 2-6 carbon atoms on a carbon chain, mainly including acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid and valeric acid.
[0129] In some embodiments, the lipid provided herein is vegetable oil.
[0130] As used herein, the term vegetable oil is a compound formed by estérification of an unsaturated fatty acids and a glycerol. The vegetable oil may be oil obtained from fruits, seeds and germ of plants, such as peanut oil, soybean oil, linseed oil, castor oil, râpeseed oil, etc. A main component of the vegetable oil is an ester generated by a linear higher fatty acid and a glycerol. In addition, the vegetable oil may further include vitamins E, K, minerais such as calcium, iron, phosphorus, potassium, fatty acids, etc.
[0131] In some embodiments, the vegetable oil provided herein is olive oil, tea oil, rapeseed oil, peanut oil, soybean oil, corn oil, safflower oil, groundnut oil, sunflower seed oil, canola oii, walnut oil, almond oil, avocado oil, castor oil, coconut oil, cottonseed oil, rice bran oîl, sesame oil, refmed palm oil or a mixture thereof.
[0132] In some embodiments, the lipid provided herein is a fatty acid, a fatty acid ester, a fatty alcohol, a lipoid, a paraffm or a mixture thereof.
[0133] In some embodiments, the lipid provided herein is a fatty acid ester. In some embodiments, the fatty acid ester provided herein is a glyceride, an ethylene glycol ester, a propylene glycol ester or a mixture thereof. In some embodiments, the fatty acid ester provided herein is a monoester, a diester, a triester or a mixture thereof. In some embodiments, the fatty acid ester provided herein is glycerides of octanoic acid and/or decanoic acid. In some embodiments, the lipid provided herein is mono-/di-glycerides of octanoic/decanoic acid or medium-chain triglycérides.
[0134] As used herein, the term medium-chain triglycéride (MCT) refers to triglycérides of fatty acids with a length of 6 to 12 carbon atoms (including one or more of hexanoic acid, octanoic acid, decanoic acid and lauric acid). The medium-chain triglycéride has a low freezîng point, is a liquid at room température and has low viscosity. In some embodiments, the medium-chain triglycéride provided herein is extracted from dry hard parts of endosperms of coconuts (e.g., Cocos mtcifera L.) or oil palms (e.g., Elaeis guineenis Jacq). A typical medium-chain triglycéride refers to a saturated octanoic acid triglycéride or a saturated decanoic acid triglycéride or a saturated octanoic acid-decanoic acid mixed triglycéride. In some embodiments, the medium-chain triglycéride provided herein meets the standards for a medium-chain triglycéride în widely accepted pharmacopoeia (e.g,, U.S, Pharmacopoeia. Pharmacopoeia of the People's Republic of China or European Pharmacopoeia). In some embodiments, the medium-chain triglycéride provided herein is MIGLYOL£812N medium-chain triglycéride.
[0135] Préparation method of pharmaceutical composition
[0136] The pharmaceutical composition provided herein may be prepared by conventional methods in the art.
[0137] In another aspect, the présent disclosure provides a method for preparing the pharmaceutical composition provided herein. The method comprises: mixing the micromoiecule PI4KIIIa inhibitor and the pharmaceutically acceptable carrier to obtaîn a mixture.
[0138] In some embodiments, the method comprises: mixing the micromoiecule PI4KIIIa inhibitor and the pharmaceutically acceptable carrier through a mechanical force. In some embodiments, the mechanical force is stirring, dispersing, shaking or ultrasonic treatment. In some embodiments, the action time ofthe mechanical force is 5 hours, 4 hours, 3 hours, 2 hours, 1 hour, 50 minutes, 40 minutes, 30 minutes, 20 minutes or 10 minutes, or a range between any two time points mentioned above. In some embodiments, heating is performed simultaneously in the mixing process. In some embodiments, the heating température is 30-80°C, 35-80°C, 40-80°C, 40-70°C, 40-60°C, 45-55°C or 55°C.
[0139] In other embodiments, the method comprises: mixing the micromoiecule PI4KIIIa inhibitor and the pharmaceutically acceptable carrier after melting the pharmaceutically acceptable carrier by heating.
[0140] In some embodiments, the method further comprises: filtering the mixture. In some embodiments, the undîssolved micromoiecule ΡΙ4ΚΙ1Ια inhibitor is removed by the filtering. In some embodiments, a filtering device used in the filtering substantially does not adsorb the micromoiecule PI4KIIIa inhibitor, for example, it adsorbs less than about 1%, 2%, 3%, 5%, 8%, 10%, 12%, 15% or 20% of the micromoiecule PI4KIIIa inhibitor in the mixture. [0141] Disease treatment method and medical use
[0142] Another aspect of the présent disclosure relates to a method for treating a
PI4KIIIa-related disease in a subject, The method comprises administrât!ng the pharmaceutical composition provided herein to a subject in need thereof.
[0143] In certain embodiments, the pharmaceutical composition provided herein includes a therapeutically effective amount of the micromoiecule ΡΙ4ΚΙΠα inhibitor [0144] As used herein, the terni therapeutically effective amount refers to an amount of médicaments that may alleviate or eliminate a disease or symptom of a subject or may prophylactically inhibit or avoîd the occurrence of the disease or symptom. The therapeutically effective amount may be an amount of médicaments that may alleviate one or more diseases or symptoms of a subject to a certain degree; an amount of médicaments that may partially or completely restore one or more physiologicai or biochemical parameters related to causes of the diseases or symptoms to normal; and/or an amount of médicaments that may reduce the possibility of occurrence of the diseases or symptoms.
[0145] A therapeutically effective dose of the micromoiecule PI4KIIIa inhibitor provided herein dépends on many factors weli known in the art, such as weight, âge, past medical htstory, treatment being currently received, health status of the subject, and intensity, allergy, hypersensitivity and side effects of médicament interaction, as well as administration routes and degree of disease development. Those skilled in the art (e.g., doctors or veterinarians) may reduce or increase the dose according to these or other conditions or requirements accordingly.
[0146] As used herein, the term subject may include a human and a non-human animal. The non-human animal includes ail vertebrates such as a mammal and a non-mammal. The subject may also be a faim animal (e.g., a cow, a pig, a sheep, a chicken, a rabbit or a horse), or a rodent (e.g., a rat or a mouse), or a primate (e.g., a gonlla or a mortkey), or a domestic animal (e.g,, a dog or a cat). The subject may be male or female, or it may be of different âges. A human subject may be a Caucasian, an African, an Asian, a Semite, or other races, or a hybrid of different races. The human object may be an elder, an adult, a teenager, a child or an infant.
[0147[ In some embodiments, the subject provided herein îs an animal such as a pig, a dog, a mortkey, a cat, a mouse, or a rat, or a human.
[0148] As used herein, the term PI4KIIIa-related disease refers to diseases associated with abnormal cellular reactions mediated by a PI4KIIIa protein kinase. In some embodiments, the PI4Knia-related disease provided herein is Alzheimer's disease.
[0149) The présent disclosure further relates to use of the phannaceutical composition provided herein in the manufacture of a médicament for treating a P I4KIIIa-related disease in a subject and the pharmaceutical composition provided herein for use in treating a PI4KIIIarelated disease in a subject.
I I (0150] Examples
[0151] Example 1: Stability and solubility of PAO in different vegetable oils
[0152] 1.1 Investigation on dissolution rate of PAO in vegetable oil
[0153] Supersaturated PAO vegetable oil solutions were respectively formulated.
allowed to stand at room température, and sampled at different tîme points to measure the dissolution rates
[0154] Table 1 : Formulation composition of PAO in vegetable oil in experiment of dissolution rate
Formulation
Names of raw and auxiliary materials PAO Vegetable oil
Ratio 1 100
Theoretical weight 50 mg 5 g
[0155] Methods: 50 mg of PAO was respectively weighed and placed into 40 ml vials, and 5 g of corresponding vegetable oil (soybean oil, sesame oil and tea oil) was respectively added. The mixture was stirred on a magnetic stirrer, and sampled at 0.5 h, 1 h, 2.5 h, 4 h and 24 h, respectively. After centrifugal filtration (12,000 rpm), the content was measured by HPLC,
[0156] The conditions of the HPLC are shown în Table 2 below:
[0157] Table 2: HPLC conditions
Chromatographie column Waters XBridge Cl 8, 5pm 4.6*250 mm
VVavelength: 214 nm Column oven 25°C
Flow rate 1.0 mL/min
Mobile phase A: 0.5% TFA aqueous solution
B; MF.OHACN-l l
Gradient elution Time (min) B%
0 10
7 10
30 80
35 80
40 100
50 100
50.1 10
55 10
[0158] Unless otherwise specified, the HPLC conditions in ail examples of the présent disclosure are the same as those mentioned above.
[0159] Results:
[0160]
Table 3: Results of 24 h dissolution rates for three vegetable oîls
Matrix Time Concentration (mg/ml)
Tea oil 0.5 h 2.51
1 h 3.97
2.5 h 3.81
4 h 3.87
24 h 3.76
Sesame oîl 0.5 h 3.48
1 h 4.33
2.5 h 4.11
4 h 4.58
24 h 4.31
Soybean oil 0.5 h 3.97
1 h 4.81
2.5 h 5.01
4 h 4.97
24h 4.47
[0161] Analysis: From the above experimental results, it can be known that in the three vegetable oîls, the PAO substantîally reached the State of dissolution equilibrium at 1 h. However, the concentration of the PAO in ail the three vegetable oîls decreased to a certain 10 extent at 24 h.
[0162] 1.2 Investigation on stability of PAO in vegetable oil
[0163] Vegetable oil solutions of the PAO were respectively formulated, and allowed to stand. Samples were taken at 0 h, 2 h, 4 h, 20 h, and 48 h respectively to investigate the content and related substances.
[0164] Table 4: Formulation composition of PAO in vegetable oîl in experîment of stability
Formulation
Names of raw and auxiliary' materials PAO Vegetable oil
Ratio 1 500
Theoretical weight 20 mg 10g
[0165) Methods: 20 mg of PAO was respectively weighed and placed into 20 ml vials, and 10 g of corresponding vegetable oil (tea oil, sesame oil and soybean oil) was added. The mixture was stirred on a magnetic stirrer at room température for 30 minutes, and filtered through a 0.22 pm filter. The fiitrate was collected, aliowed to stand at room température, and diluted with isopropanol at 0 h, 2 h, 4 h, 20 h or 48 h respectively. The stability was investigated through HPLC détection.
[0166] Results:
(0167) Table 5: Experimental results of stability of PAO in vegetable oil
Matrix Time RT % Impurities (area nonnalization method) % Total impurities (area nonnalization method) Appearance Dilution fold Concentration (mg/ml) % API residue ratio
Tea oil Oh 6.622 0.326 0.33 Clearoily liquid 7.55 0.8361 -
15.545 99.674
2h 6.558 0,705 0.70 7.46 0.8505 101.72
15.574 99.295
4h 6.531 1.012 1.01 8.07 0.8711 104.19
15,539 98,988
20 h 6,422 1.798 1.80 7.87 0.7965 95.26
15,407 98.203
48 h 6.137 4.469 4.47 7.85 0.5856 70.04
14.941 95.531
7d 6.137 2.878 2.88 7.84 0.7143 85.43
15.732 97.122
Sesame oil Oh 6.586 1.482 1.58 Clear oily liquid 7.75 0.8868 -
15.549 98.422
29.684 0.096
2h 6.527 4.616 5.27 7.82 0.7045 79.44
15.571 94.726
27,957 0.162
29.681 0.496
4 h 6.491 8.065 9.55 Turbid. witii production of particles 7,78 0.4931 55.60
15.525 90.451
27.918 0.663
29,638 0.822
20 h 6.405 22.225 31.15 7.91 0,1952 22.01
15,410 68.849
20.860 1.025
27,805 6.829
29.528 1.072
Soybean oil Oh 6.622 0.316 0.32 Clear oily liquid 8.27 1.3081 -
15.960 99.684
2 b 6.576 0.318 0.32 8.20 1.3315 101.79
15.573 99.682
4 h 6.587 0.322 0.32 8.14 1.2483 95.43
15.558 99.678
20 h 6.43 1 1.366 1.37 8.41 1.1416 87.27
15,406 98.634
48 h 6.622 8.492 8.49 7.52 0.7208 55.10
15,960 91,508
[0168] Analysis: The PAO was very unstable in the sesame oil, the total impurities increased to 9.95% after 4 h, and turbidity appeared at 20 h. In the soybean oil and the tea oil, the single impurities increased to 8.492% and 4.469% respectively after 48 h.
[0169] 1.3 Investigation on stability of PAO in végétable oïl after addition of antïoxidant
[0170] The stabilities after adding two different antioxidants (2,6-di-tert-butyl-4merhylphenol, vitamin E) to the tea oil and soybean oil including the PAO and after mixing the PAO with the vitamin E alone were investigated respectively.
[0171] Table 6: Formulation composition of PAO after addition of antïoxidant in expert ment of stability
Lot nurnber Fl F2 F3 F4
Names of raw and auxiliary materials Amount
PAO 10 mg 10 mg 10 mg 10 mg
Soybean oil 5g 5g - -
Tea oil • • 5 g -
2,6-di-tert-butyl-4-methylphenol (BHT) 5.7 mg - 5.7 mg -
Vitamin E (VE) — 71.57 mg - 71.57 mg
[0172] Methods: The components were weighed respectively according to the above formulation, and placed into a 20 ml vial. The mixture was stirred on a constant-temperature magnetic stiirer at room température for 30 min, and filtered through a 0.22 μηι filter. The filtrate was collected, ailowed to stand at room température, and diluted with isopropanol at 0 h, 1.5 h, 18 h, 24 h and 48 h respectively. The stabilîty was investigated through HPLC détection.
[0173] Results:
[0174] Table 7; Experimental results of stabilîty of PAO in soybean oïl after addition ofBHT
Ingrédients Time RT % Measurements (area normalization method) % Total impuni ies (area normalization method) Appearance Dilution fold Concentration (mg/ml) % API residue ratio
Fl (PAO+soybean oil+BHT) Oh 6.436 0.321 0.32 Clear oily liquid 8.17 1,0816 -
16.343 99.679
1.5 h 6.419 0.299 0.30 7.26 1.0248 94.75
16.300 99,702
18h 6.265 0.980 0.98 7.13 1.0383 96.00
15.927 99.020
24 h 6.246 1.0698 1.07 7.43 1.0326 95.47
15.892 98.9302
48 h 6.188 1.366 1.37 8.00 0.9592 88.68
15.819 98.634
4d 6.133 2,714 2.27 7.16 0.8146 75.31
15.742 97.286
[0175] Table 8: Experimental results of stabilîty of PAO in soybean oil after addition ofVE
Ingrédients Time RT % Measurements (area normalization method) % Total impurities (area nonnalization method) Appearance Dilution fold Concentration (mg/ml) % API residue ratio
F2 Oh 6.416 1.002 1.00 Clear oily 7.72 0.9619 -
(PAO-soybean oil+VE) 16.335 98.998 liquid
1.5 h 6.381 1.899 1.90 7.65 0.8864 92,15
16.278 98.101
18 h 6.208 68.605 68.60 Turbid. with production of particles 7.44 0.0911 9,47
15.933 31.396
24 h 6.200 88,3043 88.30 7.69 0.0569 5.92
15.886 11.6957
[0176] Table 9; Experimental resuits of stability of API in tea oil after addition of
BHT
Ingrédients Time RT % Measurements (area nonnalization method) % Total impurities (area nonnalization method) Appearance Dilution fold Concenuation (mg/ml) % API residue ratio
F3 (PAO+tea oiI+BHT) Oh 6.173 0.363 0.36 Clear oily liquid 8.45 0.9485 -
15.931 99.637
1.5 h 6.178 0,544 0.54 7.84 0.9411 99.21
15,933 99.456
18 h 6.120 1.153 1.15 7,46 0.8750 92.25
15.848 98.847
24 h 6.222 1.2143 1.21 7.96 0.8967 94.54
15.901 98.7857
48 h 6.178 1.203 1.20 8.13 0.8513 89.75
15.823 98.797
4d 6.125 1.7 il 1.71 7.90 0.8445 89.04
15,713 98.289
Table 10: Experimental resuits of stability of API in tea oil after addition of
[0177]
VE
Ingrédients Time RT % Measurements (area nomialization method) % Total impurities (area nonnalization method) Appearance Dilution fold Concentration (mg/ml) % API residue ratio
F4 (PAO+tea oil+VE) Oh 6,186 0.436 0.44 Clear oily liquid 8.19 0.8530 -
15,938 99.565
1.5 h 6,168 0.709 0.71 7.74 0.8002 93.81
15.916 99.291
18 h 6.125 2.334 2.33 7,82 0.6844 80.23
15,844 97.666
24 h 6.213 2.5903 2.59 7.87 0.7055 82.70
15.898 97.4097
48 h 6.171 3.464 3.46 8.04 0.5860 68.70
15.825 96.536
4d 6.118 9.020 9.02 7.89 0.4913 57.6
15.720 90.980
[0178] Analysis: Compared with the results without the addition of antîoxidants, dégradation of the PAO was improved after the antîoxidants were added. The efïect of adding BHT was better than that of adding VE, but from the perspective of the PAO content, there was still a significant réduction.
[0179] Example 2: Solubility and stability of PAO in mono-/di-glycerides of octanoic/decanoic acid (MCM) and medium-chain triglycérides (MCT)
[0180] 2,1 Investigation on stability of API in mono-/di-glycerides of octanoic/decanoic acid
[0181] Formulation:
[0182] Table 11: Formulation of PAO in mono-/di-glycerides of octanoic/decanoic acid in experiment of stability
Names of raw and auxiliary materials Weight
PAO 10 mg
Mono-/di-glycerides of octanoic/decanoic acid (MCM) 5g
BHT 8 mg
[0183] Methods:
[0184] 1. The raw and auxiliary materials were weighed respectîvely accordîng to the above formulation, and placed into a 20 ml vial. The mixture was stirred magnetically at room température for 30 min.
[0185] 2. After stirrîng, the mixture was filtered through a 0.22 pm nylon millipore filter with a diameter of 25 mm. The filtrate was detected by HPLC for the content and related substances.
[0186] 3. The sample was placed at room température, away from light, and sampled and detected on day 2, day 5 and day 11.
[0187] Results:
[0188] Table 12: Experimental results of 11-day stability of PAO in mono-,diglycerides of octanoic decanoic acid
Formulation composition Time RT % Measurements % Total impurities Appearance Concentration (mg/ml)
PAOBH1 +MCM Od 16.582 100.000 0.00 Clear oîly liquid 100.1400
2d 16.582 100.000 0.00 Clear oîly liquid 101.4020
5d 15.622 100.000 0.00 Clear oîly liquid 101.9920
11 d 6.546 0.247 0.25 Clear oily liquid 100.3560
15.812 99.753
|0189] Analysis: The experimental results are shown in Table 12. No reiated substances were detected in the first 5 days, and the content remai ned substantially unchanged. By 11 d, the content still did not change signifîcantly, but the reiated substance phenylarsonic acid increased to 0.25%.
[0190] 2.2 Investigation on stability of PAO in medium-chain triglycérides
[0191] The stability of the PAO in medium-chain triglycérides (MCT) was investigated.
[0192] Formulation:
[0193] Table 13: Formulation of PAO in medium-chain triglycérides in experiment of stability
Names of raw and auxiliary materîals Theoretical weight
PAO 20 mg
MCT 6.98 g
BHT 8 mg
[0194] Methods:
[0195] 1. The raw and auxiliary materîals were weighed respectively according to the above formulation, and placed into a 20 ml vial. The mixture was stirred at room température for 30 min.
[0196] 2. After stirring, the mixture was filtered through a 0.22 pm nylon millîpore fi lier with a diameter of 25 mm. The filtrate was detected by HPLC for the content and reiated substances.
[0197] 3. The sample was placed at room température, away from light, and sampled and detected on day 5 and day 14.
[0198] Results:
[0199] Table 14; Experimental results of 14-day stability of PAO in medium-chain triglycérides
Formulation composition Time RT % Measurements % Total impurities Appea rance Weight (mg) Volume (ml) Concentration (mg/ml) % Content
PAO- BHT+ MCT Od 6.552 0.083 0.29 Clear oily liquid 502.2 5.00 100.4400 0.2347
15.591 99.709
31.121 0.208
5d 6.576 0,196 0.40 Clear oily liquid 500.05 5.00 100.0100 0.2418
15,851 99.601
31,453 0.202
14 d 6.493 0,331 0.52 Clear oily liquid 508.17 5.00 101.6340 0.2222
15,676 99.481
31.358 0.189
[0200] Analysis: From the above experimental results, it can be known that the total related substances of the sample were 0.29% on day 0, increased to 0,40% on 5 d, and increased to 0.52% on day 14. The content of the phenylarsonic acid impurity (phenylarsonic 10 acid) at rétention time of 6.55 min was 0.083% on day 0, and increased to 0.33% after 14 days.
[0201] 2.3 Investigation on stability of PAO in MCM and MCT solutions without addition of antioxidant BHT and influence of stirring time on dissolution
[0202] Formulation:
[0203] Table 15; Formulation of PAO in MCT and MCM without addition of antioxidant BHT in the investigation on stability
Lot number F13-180426 F14-180426
Nantes of raw and auxiliary materials Theoretical weight Theoretical weight
PAO 40 mg 20 mg
MCT 20 g
MCM - 10g
[0204]
Processes;
[0205] F13-180426
[0206] 1, The raw and auxiliaiy materials are weighed according to the above formulation, and placed into a 10 ml vial.
[0207] 2, The mixture was stirred in a constant-temperature magnetic stirrer for 30 min. Approxîmately 10 g was taken, and filtered through a 0.22 gm millipore filter membrane with a diameter of 25 mm.
[0208] 3. The remainîng part was continuously stirred for 1 h, 2 h and 4 h, and then sampled. The samples were filtered through a 0.22 pm millipore filter membrane with a diameter of 25 mm.
[0209] 4. The filtrâtes were detected by HPLC respectively.
|0210] F14-180426
[0211] 1. The raw and auxiliary materials are weighed according to the above formulation, and placed into a 10 ml vial.
|0212[ 2. The mixture was stirred in a constant-temperature magnetic stirrer for 30 min.
[0213] 3. The mixture was filtered through a 0.22 μιη millipore filter membrane with a diameter of 25 mm. The filtrate was detected by HPLC.
[0214] Results:
[0215] Table 16: The dissolution under different stirring times and stability' results for 35 d room-temperature placing of F13 (PAO+MCT)
Formulation composition Time RT % Measurements % Total impurities Appearance Weight (mg) Volume (ml) Concentration (mgTnl) % Content % Content relative to 0 cl
Fi 3-1 SO 42 6 (PAO+ MCT) Od tO.S h) 14.712 99.784 0.22 Clear oily liquid 757.95 5.00 151.5900 0.1944 -
30.670 0.216
Odl h 14.657 99.783 0.22 Clear oily liquid 753.13 5.00 150.6260 0.1963 100.98
30.632 0.218
2h 14.611 99.789 0.21 Clear oily liquid 761.95 5.00 152.3900 0.1999 102.83
30.626 0.211
4h 14.517 99.780 0.22 Clear oily liquid 758.61 5.00 151.7220 0.1998 102.78
30.571 0.220
6d 6.407 0.159 0.32 Clear oily liquid 748.96 5.00 149.7920 0.1930 99.28
15.485 99.682
31.208 0.159
Il d 6.471 0.121 0.30 Clear oily liquid 755.72 5.00 151.1440 0.1971 101.39
15.617 99.699
31.302 0.180
15 d 6.223 0.168 0.34 Ciear oily liquid 761.07 5.00 152.2140 0,1947 iœ.15
15.080 99.665
31.030 0.167
21 d 5.948 0.140 0,30 CJear oily liquid 754.03 5.00 150.8060 0,1933 99,43
14.479 99.699
30.632 0.161
27 d 6.133 0.0816 0.29 Clear oily liquid 757.76 10 75.7760 0.2024 104.12
14.778 99.708
30.741 0.138
31.752 0.073
35 d 6.652 0.2059 0.41 Clear oily liquid 761.96 10 76.1960 0,1923 98,92
15.976 99.593
31.527 0.147
32.476 0.054
[0216] Analysis: From the above experimental results, it can be known that along with the extension of the stirring time, the content of the API in MCT substantially tended to be stable and overall approximated the theoretical content, namely 0.2% (w/w). The détection 5 results on 0 d showed that no phenylarsonic acid impuritîes were produced, and only impurîties of the active pharmaceutical ingrédients themselves appeared near 31 min. With the continuation of room-temperature placîng, the phenylarsonic acid at an amount of 0.159% began to appear from 6 d. The content of phenylarsonic acid at each subséquent time point fluctuated around the détection resuit on 6 d. It can be seen that after the API was placed in oil for a period of time, the phenylarsonic acid content tended to be stable.
[0217] Table 17; Détection results of stability ofF14 (PAO + MCM) for placement at room température for 35 d
Formulation composition Time RT % Measurements % Total impurities Appearance Weight (mg) Volume (ml) Concentration (mg'ml) % Content % Content relative to 0 d RSD
F14-180426 (PAO+ MCM) Od 14.345 99.809 0.19 Clear oily liquid 752.58 5.00 150.5160 0.1930 - -
30.465 0.191
6d 6.35 0.116 0.26 Clear oily liquid 754,45 5,00 150.8900 0.1857 96.22 -
15,374 99.742
31,110 0,142
11 d (taking upper layer) 6.391 0.144 0.26 Clear oily liquid 759,10 5.00 151,8200 0,1924 99.69 0,86
15,432 99,741
31.171 0.115
11 d ( taking lower layer) 6.395 0.157 0.27 Clear oily liquid 759.70 5.00 151.9400 0.1908 98.86
15.401 99.726
31.164 0.118
1 ] d (taking after being uniformly mixed) 6.385 0.120 0.24 Clear oily liquid 754.27 5.00 150.8540 0.1941 100.57
15.381 99.759
31.154 0.121
15 d 6.188 0.149 0.27 Clear oily liquid 759.08 5.00 151.8160 0.1861 96.42 -
15.019 99.735
30.967 0.116
21 d 5.906 0.144 0.46 Clear oily liquid 763.45 5.00 152.6900 0.1905 98.70 -
14.397 99.540
30.596 0.316
27 d 6.04 0.144 0.07 Clear oily liquid 756.53 10 75.6530 0.1968 101.97 -
14.617 99.931
35 d 6.595 0.284 0.28 Clear oily liquid 744.51 10 74.4510 0.1803 93.42
15.829 99.716
Pure PAO (SP0020182-029) - 14.144 99.699 0.30 White powder 20.63 20.00 1.0315 102.8 - -
30.401 0.209
33.661 0.047
35.920 0.046
[0218] Analysis: For F14 taking MCM as a solvent, the overall trend of the stability was consistent with that of F13. Phenylarsonic acid began to appear from 6 d, was in a relatively stable state, and reached the maximum on 35 d.
[0219J 2,4 Investigation on influence factors (high température, high humidity and light exposure) of F15 and F16
[0220] F15 and F16 were placed under high température (50°C), high humidity (92.5%
RH) and light exposure (4,500 ix) respectively. Samples were taken on 5 d, 10 d and 30 d respectively to detect the content and related substances.
[0221] Table 18: Formulation of F15 for influence factor investigation
Lot number F15-18O515
Nantes of raw and auxiliary materials Theoretical weight
PAO 100 mg
MCT 50 g
[0222] Table 19; Formulation of F16 for influence factor investigation
Lot number FI6-180515
Names of raw and auxiliary matenals Theoreticai weight
PAO 100 mg
MCM 50 g
[0223] Methods:
[0224] Fl 5-]80515: The active pharmaceutical ingrédients were passed through a
200-mesh screen. The raw and auxiliary materials were weighed, and placed into a vial. The mixture was magnetically stirred for 30 min, and filtered through a 0.22 pm millipore filter of 25 mm. The solution was taken triplicate, and placed under the conditions of high température of 50°C, high humidity of 92.5% RH and light exposure of 4,500 Lx respectively. Samples were taken on 5 d. 10 d and 30 d respectively to investigate the influence factors, [0225] F16-180515: The MCM was heated in a water bath at 40°C for 3-5 min until the MCM was melted into a liquid, and the remaining steps are the same as those of F15 to perform investigation on the influence factors,
[0226] Results of the influence factors of F15-180515 are shown in Tables 20-23; [0227] Table 20: Analysis results of F15 in refrigerator (2-8°C) on 5 d, 10 d and 33 d
Naine Time RT RRT % Measurements % Total impurities Appea rance Weiabt (mg) Volume (ml) Concentration (nig/m1) % Content % Content relative to 0 d
F15-180S15 (MCT+PAO) Low température (2-8°C) Od 14.345 1.00 99.794 0.206 Clear 01 ly liquid 759.00 10 75.9000 0.1544 -
30.584 2.13 0,206
5 d 6.339 0.42 0,11 0.37 761.78 10 76.1780 0.1635 105.89
15.217 1.00 99.629
31.054 2.04 0.261
lOd 6.596 0.42 0.275 0,47 768.23 10 76.8230 0.1620 104.92
15.802 1.00 99.535
31.479 1.99 0.190
33 d 15.428 1.00 99.885 0.12 757.02 10 75.7020 0.1545 100.06
31.264 2.03 0.115
Note: The 0 d results were measured on the day of sample formulation, and were the sanie data as other influence factors The parts marked in red are for the impurtry phenylarsonic acid.
[0228] Table 21 : Analysis results of F15 in high-humidity (92.5% RH) stability chamber on 5 d, 10 d and 32 d
Na me Time RT RRT % Measurements % Total impurities A pp va rance Weight (mg) Volume (ml) Concentration (mg/ml) % Content Relative content per cent âge
Relative to 0 d Relative to 2-8'C
F15-180515 (MCT+ PAO) High humiditv (92.5% RH) 5d 6.195 0,42 0.091 5.02 Clear oily liquid 750.85 10 75.0850 0.1630 105.57% 99.69%
14.875 1,00 94.576
30.795 2.07 0.252
31.857 2.14 0.404
32,838 2.21 4.676
10 d 6.453 0.42 0.1589 1.34 748.56 10 74.8560 0.1626 105,31% i 00.37%
15,467 1.00 98.834
31.218 2.02 0.258
32.190 2,08 0.131
33.180 2.15 0.918
33 d 6.522 0.41 0.5846 0.85 746.06 10 74.6060 0.1547 100,19% 100.13%
15.721 1.00 99.148
31.376 2.00 0.146
32.339 2.06 0.122
Note: The relative content percentages were respectively relative to the day of sample formulation and storage in die refrigerator for the same time period. The parts marked in red are for die impurity phenylarsonic acid.
[0229] Table 22: Analysis resuIts of F15 in (50°C) stability chamber on 5 d, 10 d and 32 d
Na me Time RT RRT % Measurements % Total impurities Appea rance Weight (mg) Volume (ml) Concentration (mg/ml) % Content Relative content perce otage
Relative to 0 d Relative to 2-8°C
Fl 5-180515 (MCT+PAO) High température (50°C) 5 d 6.287 0,42 Û.1381 0.40 Clear oily liquid 750.64 10 75.0640 0.1620 104.92% 99.08%
15.130 1,00 99.603
30.986 2.05 0.259
10 d 6.52 0.42 0.3304 0.59 757,43 10 75.7430 0.1623 105,17% 100.19%
15.698 1.00 99.410
31.383 2.00 0 209
32.357 2.06 0.051
33 d 6.631 0.42 0,4232 0,51 747.50 10 74.7500 0.1572 101.81% 101.75%
15.917 1.00 99.491
31.543 1.98 0.086
Note: The relative content percentages were respectively relative to the day of sample formulation and storage in die refrigerator for die same time period. The parts marked in red are for the impurity phenylarsonic acid.
[0230] Table 23: Analysis results of F15 in light exposure (4,500 Ix) stability chamber on 5 d, 10 d and 32 d
Xante Time RT RRT % Measurements % Total impurirle* Appea rance Weight (mg) Volume (ml) Concentration (mg/ml) % Content Relative content percentage
Relative to 0 d Relative to 2-8'C
F154 80515 (MCT+ API) Light expos tire (4,500 Ix) 5d 6.2 0.41 3.4358 5.32 Clear oily liquid 764.03 10 76.4030 0.1395 90.35% 85.32%
15.003 1.00 94.680
21.766 1.45 1.825
30.859 2.06 0.059
lOd 6.43 0.41 4.8882 7.33 757.87 10 75.7870 0.1240 80.31% 7654%
15529 1.00 92.671
22.209 1.43 2.441
33 d 6548 0.41 7.2746 25.45 761.60 10 76.1600 0.0382 24.7490 24.72%
15.823 1.00 74550
22.487 1.42 37.975
27.802 1.76 0.201
Note: The relative content percentages were respectively relative to the day of sample formulation and storage in the refrigerator for the same rime period. The parts marked in red are for the impurity phenylarsonic acid.
[0231] Analysis: From the results, it can be known that the contents of Fl5 under low-temperature conditions on 5 d and 10 d were higher than those on 0 d (the day of sample formulation), and the impurity phenylarsonic acid was detected. The content resuit on 33 d 5 was comparable to that on 0 d, and no phenylarsonic acid was detected.
[0232] The change trend of the API content under high-temperature and highhumidity conditions was consistent wîth that at low température. The phenylarsonic acid began to appear from 5 d. Under high-temperature and high-humidity conditions, the content of the phenylarsonic acid reached 0.42% and 0.58% respectively on 33 d. Under high- humidity conditions, new unknown impurities appeared after the rétention time of 30 min. Under high-temperature conditions, similar impurities also appeared on 10 d, and the content L was unstable. ‘
[0233] Under light exposure conditions, the API degraded rapidly. On 33 d, the API content decreased to 24.72%, and the phenylarsonic acid content increased to 7.72%. In addition, a new unknown impurity (at the rétention time of 22 min) began to appear from 5 d, and the impurity increased rapidly and increased to 17.98% on 33 d. At the same time, another new unknown impurity (at the rétention time of 27.8 min) began to appear on 33 d. [0234] Conclusions: After F15 was placed under varions conditions for 33 d, the content of F15 changed to a certain extent under low-temperature, high-temperature and high-humidity conditions. The trends for the three conditions were the same, which first
increased and then decreased. This change may be caused by the inaccurate content profiles of reference substances.
[0235] Related substances increased by different degrees under varions conditions. The stability of the sample was poor under light exposure conditions. Along with the time extension of the placement, the API content decreased significantly, and the total impurity content increased significantly. The case at high température was comparable to that at high humidity, and the impurities increased slowly. For placing under high-température conditions for 5 d, only 0.138% of the phenylarsonic acid impurity appeared.
[0236] Results of the influence factors for F16 are shown in Tables 24-27:
[0237] Table 24: Analysis results of F16 in réfrigérator (2-8’C) stability chamber on 5 d, 10 d and 33 d
Naine Time RT RRT % Measurements % Total impurities Appearance Weight (mg) Volume (ml) Concentration (mg/ml) % Content Content percentage relative to 0 d
F16-180518(MCM+PAO) Low température (2-8°C) Od 14.121 1.00 99.794 0.206 Clear oily liquid 754.61 10 75.4610 0.1429 -
30.420 2.15 0.206
5 d 14.645 1.00 100.000 - 754.57 10 75.4570 0.1508 105.53%
10 d 15.232 1.00 100.000 752.64 10 75.2640 0.1492 104.41%
33 d 6.269 0.41 0.526 0.53 760.16 10 76.016 0.1426 99.79%
15.168 1.00 99.474
Note: The relative content percentages were respectively relative to the day of sample formulation and storage in the refrigerator for the saine time period. The parts marked in red are for the impurity phenylarsonic acid.
[0238] Table 25: Analysis results of F16 in high-humidîty (92.5% RH) stability chamber on 5 d: 10 d and 32 d
N a me Time RT RRT % Measurements % Total impurities Appearance Weight (mg) Volume (ml) Concentration (mgnil) % Content Relative content percentage
Relative to û d Relative to 2-8°C
1-16-18()51 S(MCM+ PAO) High humidity (92.5% RII) 5 d 14.611 1.00 100.000 - Clear oily liquid 757.40 10 75.7400 0.1660 116.17% 110.08%
lOd 14.611 1.00 100.000 - 747.76 10 74.7760 0.1649 115.40% i 10.52%
33 d 6.335 0.41 0.789 0.79 754.27 10 75.4270 0.1503 105.18% 105.40%
15.367 1.00 99.211
Note: The relative content percentages were respectively relative to the day of sample formulation and storage in die refrigerator for the same time period. The parts marked in red are for the impurity phenylarsonic acid.
[0239] Table 26: Analysis results of F16 in high-température (50°C) stability chamber on 5 d, 10 d and 32 d
Naine Time RT RRT % Measurements % Total impurities Appearance Weight (mg) Volume (ml) Concentration (mg/ml) % Content Relative content percentage
Relative to 0 d Relative to 2-8'C
F16-180518(MCM+ PAO) High température (50°O 5 d 6.043 0.41 0.0654 0.07 Clear oily liquid 762.33 1Ü 76.2.330 0.1658 116.03% 109.95%
14.631 1.00 99.935
lOd 6.301 0.41 0215 022 761.63 10 76.1630 0.1647 11526% 110.39%
15212 1.00 99.785
33 d 6.35 0.41 1.0396 1.04 757.74 10 75,7740 0,1504 10525% 105.47%
15.397 1.00 98.960
Vote; The relative content percentages were respectîvely relative to the day of sample formulation and storage in the réfrigérator for the same time period, The parts marked in red are for the impurily phenylarsonic acid.
[0240] Table 27; Analysis results of F16 in light exposure (4,500 LX) stability chamber on 5 d, 10 d and 32 d
Name Time RT RRT % Measurements % Total impurities Appearance Weight (mg) Volume (ml) Concentration (mg/m!) % Content Relative content percentage
Relative to 0 d Relative to 2-8’C
F16-180518(MCM+ API) light exposure (4,500 LX) 5 d 6.011 0.41 0.3294 0.33 Clear oily liquid 752.02 10 752020 0.1605 112.32% 106.43%
14.622 1,00 99.671
lOd 627 0,41 0.5011 0.50 753.05 10 75.3050 0.1587 111,06% 106,37%
15200 1.00 99.499
33 d 6.334 0.41 3.1528 3,15 760.93 10 76.0930 0,1388 97.13% 97.34%
15.377 1,00 96,847
Note: The 0 d results were measured on the day of sample formulation, and were the same data as other influence factors, The parts marked in red are for the impurily phenylarsonic acid.
[0241] Analysis: In terms of reiated substances, the only impurity in F16 under varions conditions was phenylarsonic acid. Because the MCM auxiliary material itself had a set of solvent peaks after the rétention time of 30 min under this HPLC method, which overlapped with those of the impurities of the API in this area, the impurities of the API near here were not reflected in the table. After F16 was placed under low-temperature, high10 humidity and high-temperature conditions for 33 d, the phenylarsonic acid increased more sîgnifîcantly than that in F15, reaching 0.53%, 0.79% and 1.04% respectîvely. Under light exposure conditions, the content of phenylarsonic acid in F16 was far lower than that in F15, and the main dégradation impurity in F15 at the rétention time of 22.48 min did not appear in F16.
[0242] In ternis of the API content, the change trends under high-température and high-humidity conditions were the same. At each time point, the API content was higher than that under low-temperature conditions, and also higher than the 0 d détection resuit. On 0 d, because the formulated sample w as placed in the refrigerator for severai hours before being detected, the API content detected on 0 d was consistent with that at low température. Since MCM was solid at low température, it needed to be melted into a liquid before sampling during room-temperature détection. Therefore, the reason for the low content may be that the API was not completely re-dissolved in the MCM in the freezing and thawing process of the API in MCM solution, thus resulting in the low API content. Under light exposure conditions, the content of API relative to 0 d graduai ly decreased.
[0243] Conclusions: In general, Fl5 was less stable than F16 under light exposure conditions, but more stable than F16 under other influence factor conditions.
[0244] 2.5 Placement stabilîty forF15 and F16 (40°C/75% RH)and PAO in glyceryl monolinoleate (MAISINE CC) (40°C/75% RH and room température)
[0245] F15 and F16 were placed at 40°C/75% RH to investigate the stabilîty. At the same time, a PAO in glyceryl monolinoleate solution was placed at 40°C/75% RH and at room-temperature, and samples were taken at different time points to investigate the stabilîty. [0246] Table 28: Formulation of F15 and F16 and PAO in MAISINE CC solution for placement stabilîty
Lot number F15-180601 F16-180601 F18-180601
Names of auxiliary materials Theoretical weight Theoretical weight Theoretical weight
PAO 20 mg 20 mg 30 mg
MCT 10g -
MCM - 10 g -
MAISINE CC - - 15 g
[0247] Methods:
[0248] F15-180601 : The active pharmaceutical ingrédients were passed through an 80-mesh screen. The raw and auxiliary materials were weighed, and placed into a vial, The mixture was stirred on a magnetic stirrer at room température for 0.5 h, and filtered through a 0.22 pm nylon millipore filter. About 7 g of the filtrate was weighed, placed into a vial and then put into a 40°C/75% RH stabilîty chamber. Samples were taken at different time points
to investi gâte the stability. The remaining part of the filtrate was placed into a vrai and then put into a 25°C/60°C stability chamber for later use.
[0249] F16-180601 : The MCM was wreighed, placed into a vial and then melted into a liquid in a w'ater bath of 40°C, and the remaining steps are the same as those of F15-180601.
[0250] F18-180601 : The formulation method was the same as that of F15 -180601.
The filtrate was divided into two parts, one part was placed in the laboratory, away from iight and the other part was placed in a 40°C/75% RH stability chamber. Samples w-ere taken at different time points to investigate the stability'.
[0251] Results:
[0252] Table 29: Analysis results of content and related substances of F15 in (40°C/75% RH) stability chamber within 33 d
Name Time RT RRT % Measurements % Total impur! fies Appearanee Weight (mg) Volume (ml) Concentration (m g/m 1) % Content Content percentage relative to 0 d
F151S0601 (MCT+ PAO) 0 d 15.800 1.00 99.6146 0.39 Ciear oily liquid 761.06 10 76.1060 0.1472 -
31.423 1.99 0.3247
32.363 2.05 0.061
5 d (40ûC/75% RH?) 15.355 0.49 99.838 0.16 754.20 ίο 75.4200 0.1595 108.36
31.118 1.00 0.162
13 d (40=075% RH) 6.605 0.42 0.281 0.45 746.92 10 74.6920 0.1515 102.92
15.841 1.00 99.555
31.498 1.99 0.164
31 d (40°C/75% RH) 6.259 0.41 0.237 0.41 750.18 10 75.0180 0.1532 104.08
15.184 1.00 99.586
30.581 2.01 0.177
Note; The parts marked in red are for the impurity phenylarsonic acid.
[0253] Table 30: Analysis results of content and related substances of F16 in (40°C/75% RH) stability chamber within 33 d
Nanie Time RT RRT % Measu remonts % Total impurities Appearanee Weight (mg) Volume (ml) Concentration (mg/ml) % Content Content percentage relative to Od
ΓΙΟ- Ι 80601 (MCM +PAO) 0 d 15.621 1.00 100.000 - Ciear oily liquid 746.99 10 74.699 0.1331 -
5 d (40JC/75% RH) 15.161 1.00 100.000 - 763.13 10 76.3130 0.1342 100.83%
13 d (40°C/75% RH) 6.517 0.42 0.458 0.46 753.17 10 75 3170 0.1339 100.60%
15.648 1.00 99.542
31 d (40-075% RH) 6.15 0.41 0,298 0.30 754.98 10 75.4980 0.1365 102.55%
15.052 1.00 99.702
Note: The parts nwked in red are for the impurity pheny Jars unie acid.
[0254] Table 31: Analysis results of content and related substances of F16 in (40°C/75% RH) stabi lity chamber and under room-temperature conditions within 5 d
Name Time RT RRT % Measurements % Total inipuriries App carence Weight (mg) Volume (ml) Concentration (mg-ml) % Content
F18180601 (Maisine /-PAO) Od 6.55 0.42 4.5601 4.56 Yellow ciear oily liquid 748.54 10 74,8540 0,0619
15.753 1.00 95.440
5 d (roomtemperature laboratory) 6.273 1.00 100.000 - 771,89 10 77.1890 -
5 d (40’075% RH) 6.267 1.00 100.000 - 753,72 10 75.3720 -
Note: The parts marked in red are for the impurity phenylarsonic acid.
[0255] Analysis: For F15, no phenylarsonic acid was detected on 0 d, but an unknown related substance appeared at the rétention time of 32 min. It was later confmned that this impurity was an external pollution impurity. In addition, the phenylarsonic acid impurity began to appear from 13 d, reaching 0.28%; and it decreased slightly after 31 d. The content of PAO was generally higher than that on 0 d and reached the maximum on 5 d, and the content relative to 0 d reached 108.36%.
[0256] For F16, the content substantially tended to be stable. The change trends of the related substances in F16 were consistent with tbose exhibited in Fl5. The phenylarsonic acid impurity began to appear from 13 d, reaching 0.46%; and it also decreased slightly on 31 d, At the same time point, the phenylarsonic acid content was higher than that of F15.
[0257] For F18 with glyceryl monolinoleate as the matrix, the PAO was extremely unstable thereîn, and 4.56% phenylarsonic acid was detected on the day of formulation. The PAO was completely degraded both at room température and in a 40°C/75% RH stabi lity chamber on 5 d.
[0258] Conclusions: Based on the comprehensive comparison stability results of F15 20 and F16 under 40°C/75% RH conditions withm 31 d, the stability of F15 was slightly higher than that of Fl6. In addition, compared with previous stability results at room température, the stability of F15 under accelerated conditions (40°C/75% RH) was comparable to that at
room température, while the placement stability of F16 under accelerated conditions (40°C/75% RH) was slightly lower than that at room température.
]0259] 2.6 Placement stability of PAO sample (25°C/60% RH and 2-8°C) |0260] PAO samples (PAO in MCT solutions, having a concentration of 1.5 mg/ml) were stored under 25°C/60% RH (accelerated) and 2-8°C (long-term) conditions for 6 months respectively. The HPLC test resuIts of the stability are shown in Table 32 and Table 33. The HPLC analysis method and parameters are substantially the same as those in Table 2, except that the mobile phase A is changed from 0,05% TFA aqueous solution to 0.05% HjPO4 aqueous solution.
[0261] Table 32: Accelerated stability test results of PAOsample stored under
25°t760% RH conditions for 6 months
Test itéra Method N/A Time (month)
0 1 2 3 6
Properties GAM-GP-QC-012 Colorless clear oily liquid Colodess clear oily liquid Colorless clear oily liquid Colorless clear oily liquid Colorless clear oily liquid
Clarity ChP <O9O2> Compilant Compilant Compilant Compilant Compli ant
Related substances AM-DCGO25-O1 Names of impurities % Impurities
RRT0.39 (0,38-0,40) ND ND ND ND ND
RRT0.45 (0.440,46): PA 0.25 022 0.24 0.27 0.11
RRT1.08(L09) ND ND ND ND N/A
RRT1.25 (1.24) ND ND ND ND ND
RRT1.37 (1.39) ND N/A N/A N/A N/A
RRT1.39 ND N/A N/A N/A N/A
RRT1.41 (1.39-1,42) ND ND ND ND ND
RRT1.48 (1.46-1.49) 0,12 0.13 0.13 0.13 0.14
RRT1.67 (1.64-1.68) 0.20 0.20 0.20 0.19 0.19
RRT1.79 (1.76-1.80) <LOQ (0.03} <LOQ (0.03) <LOQ (0.03) <LOQ (0.03) ND
RRT2.06 (2.02-2.07) ND ND ND <LOQ (0,03) ND
% Total impurities 0.57 0.54 0.57 0.60 0.44
Content AM-DCG025-0I N/A 100.1% 99.5% 100.0% 98.6% 97.7%
Microbial limit AM-P101-04 N/A Total aérobic bacteria: <102 CFU mL Mollis and veasts: <50 ' CFU'mL Escherichia coh: Not detected per 1 mL
ND: Below the détection limit (0.03%); N/A: not applicable
[0262] Table 33: Long^snnstsbiitytestresulteofPAOsample stored under 2-8=C conditions for 6 months
Test item Method N-'A Time (montli)
0 3 6
Properties GAM-GP-QC-012 Colorless clear oily liquid Colorless clear oîly liquid Colorless clear oily liquid
Clarity ChP <0902> Compilant Compilant Compilant
Reiated substances AM-DCGO25-OI Names of impurities % Impurities
RRT0.39 (0.38-0.40) ND ND ND
RRT0.45 (0.44-0.46):PA 0.25 0.15 0.10
RRT1.08 (1.09) ND ND N/A
RRT1.25(L24) ND ND ND
RRTL37(1.39) ND N/A N/A
RRT1.39 ND N/A N/A
RRT1.41 (1.39-1.42) ND ND ND
RRT1.48 (1.46-1.49) 0.12 0.14 0.13
RRT1.67 (1.64-1.68) 0.20 0.20 0.20
RRT 1.79 (1.76-1.80) <LOQ (0.03) <LOQ (0.03) ND
RRT2.06 (2.02-2.07) ND <LOQ (0.03) ND
% Total impurities 0.57 0.48 0.44
Content AM-DCGO25-01 N/A 100.1% 99.1% 98.2%
Microbial limit AM-PI01-04 N/A Total aérobic bacteria: <10- CFU/mL Molds and yeasts: <50 CFU/mL Escherichia coli: Not detected per 1 mL
ND: Below the détection limit (0.03%); N/A: not applicable
[0263] Example 3; Screening formulation conditions of PAO in MCT solution
[0264] Experiments on the influence factors of F15 showed that API was relatively stable in MCT under high-temperature (50°C) conditions, Therefore, it is considered to promote the dissolution of PAO by heating.
Table 34: Formulation for dissolution of API by heating
Lot number Fl 5-180929
Naines of auxiliary materials Theoretîcal weight
PAO 20 mg
MCT 10 g
[0265] Methods: The température of a constant-temperature magnetic stirrer was preset at 503C. After the température reached 50°C, the raw and auxiliary materials were weighed and placed into a 25 ml round-bottom flask. The mixture was stirred at a set speed of 800 rpm m the dark, and phenomena were observed at different time points. After the mixture 10 became clear, a sample was taken and filtered through a nylon millipore filter membrane with a pore size of 0.22 pm. The content and related substances were detected by HPLC.
[0266] Status of sample:
[0267] Table 35: Phenomena of PAO at different time points during dissolution by heating
Time point Phenomena
Very beginning of stiriing PAO was suspended in MCT, and the System was turbid, with a large number of obvious big particles.
Stirring for 15 min Except for a small number of visible big particles, the system appeared to be clear.
Stirring for 30 min No visible undissolved substances were found, and the System appeared to be clear and transparent.
Stirring for 1 h Phenomena were the same as those at the time point of stirring for 30 min.
Stirring for 2 h Phenomena were the same as those at the time point of stirring for 30 min.
Stirring for 4 h Phenomena were the same as those at the time point of stirring for 30 min.
[0268] Resuits:
[0269] Table 36: HPLC détection resuits of F15 at different time points during dissolution by heating
Name Sampling time point RT % Measurements % Total impurities Appea rance % Content % RSD
F15-180929 (PAO+MCT) 0.5 h 14.945 99.4157 0.58 Clear oily liquid 0.2005 0.53
30.517 0.3651
31.446 0.0462
35.520 0,1731
1 h 14.916 99.2742 0.73 0.2017
28.050 0,1021
30.488 0.3745
31.429 0.0565
35.502 0.1927
2 h 14.885 99.3120 0.69 0.1991
27.997 0.0946
30.455 0.3791
31.385 0.0539
35.482 0.1603
4 h 14.842 99.3127 0.69 0.2006
27.965 0.0894
30.413 0.3891
31.359 0.0515
35.459 0.1573
[0270] Analysis: The results are shown in Table 36. After the sample is stirred to become clear (0.5 h-4 h), the content reached the theoretical concentration (0.2%), and no phenylarsonic acid was generated. At the same time, an impurity at the rétention time of 28 5 min was newly produced after 1 h. This impurity resulted from the active phannaceutical ingrédients. Therefore, stirring and heating for 0.5 h was the optimal formulation condition.
[0271] Example 4: In vitro release experiments of PAO
[0272] Four formulations were prepared, and in vitro experiments were carried ont to simulate the release of the formulations în the stomach. Since PAO had a higher affinity with 10 proteins, a 0.1N HCl solution was temporarily used instead of artificial gastric juice. The unified formulation concentration was 2 mg/g. The release ofthe formulation was investigated through a dissolution instrument.
[0273) Table 37: Dissolution method for in vitro release experiments
Release medium/volume Speed Sampling points Température Method
0.1N hydrochloric acid 200 ml 100 ipm 15 min, 30 min, 45 min. 60 min and 120 min 37=0.5=C Paddle dissolution method
[0274] 4.1 Investigation on dissolution and stability of PAO in 0.1N HCl
[0275] 20 mg of PAO was weighed, and dissolution experiments were carried out according to the above dissolution method (repeated once for the same sample). Samples 5 were taken at different time points with a sampling volume of 3 ml, and no solution was supplemented after sampling. Each sample was subjected to 0.22 pm filtration. HPLC détection was performed on the filtrate to investigate the dissolution rate and stability. The dissolution solution at 2 h was continuously injected within 24 h to investigate the stability.
[0276] The results are shown in FIG. 1 and Tables 38-41:
[0277] Table 38; Investigation on dissolution rate of PAO through dissolution method
Sample information PAO sample 1 PAO sample 2
Time (min) Cumulative release (%)
0 0 0
15 18.77 14.17
30 35.13 24.33
45 46.68 30.92
60 55.83 0.00 (data missing due to an HPLC problem)
120 85.04 79.29
[0278] Table 39: Related substances of dissolution sample of PAO sample 1
Time RT % Area % Total impurities
15 min 5.539 0.3937 1.80
7.613 1.0269
15.841 98.1977
20.882 0.3817
30 min 5.541 0.6226 6.17
7.632 3.1118
15.864 93.8296
18.540 0.3762
20.882 2.0599
45mm 5.528 0.4429 7.34
7.606 4.3722
15.830 92.6648
18.465 0.5120
20.141 0.3500
20.89 1.6580
1 h 5.517 0.4258 7.43
7,571 4.8278
15.813 92.5727
18.443 0.3926
20.145 0.3062
20.885 1.4748
2 h 5.503 0.5 5.44
7.566 2.8557
15.805 94.5582
16.786 0.0593
18.439 0.2538
20.877 1.7730
[0279]
Table 40: Related substances of dissolution sample of PAO sample 2
Time RT % Area % Total impurities
15 min 5.54 0.7842 7.01
7.615 3.8508
15.855 92.9863
18.525 0.5621
20,966 1.8166
30 min 5.528 0.69 . 6.03
7.615 3.8168 ’
15.870 93.9682
18.547 0.2166
20.974 1.3085
45 min 5.512 0.6174 7.13
7.583 3.5237
15.819 92.6648
16.797 0.0880
18.458 0.6202
20.887 2.2803
1 h 5.498 0.9555 6.39
7.559 2.9402
15.799 93,6145
16.774 0.0634
18.433 0.4308
20.122 0.0413
20.876 1.9543
2h 5.514 0.5835 8.26
7.586 4.8660
15.817 91.7416
16.806 0.1048
17.560 0.0743
18.450 0.3442
20.885 2.2857
[0280] Table 41: 24 h (liquid injection plate) stability détection results of dissolution sampie at 2 h of PAO sample 1
Time RT % Area % Total impurities
Oh 5.514 0.5835 8.26
7.586 4.8660
15.817 91.7416
16.806 0.1048
17.560 0.0743
18.450 0,3442
20.885 2.2857
2h 5.514 0.3373 7.89
7.590 4.9222
15.820 92.1117
16.789 0.1023
18.446 0.2885
20.878 2.2380
4 h 5.528 0.3084 7.98
7.602 4.8785
15.832 92.0247
16.812 0.1277
17.580 0.0406
18.463 0.3277
20.892 2.2924
8h 5.524 0.3731 8.23
7.603 4.9485
15.843 91.7686
16.814 0.1095
17,571 0.1033
18.469 0.4172
20,891 2.2798
12 h 5.524 0,3531 7.95
7.595 4.9000
15.848 92.0488
16.837 0.1339
18.528 0.2220
20.963 2.3423
24 h 5.555 0.2967 7.84
7.664 4.8424
15.886 92.1565
16.852 0.1088
18.559 0.3403
20.961 2.2552
[0281] Analysis: In the experiment, from the time when PAO powder was added to a dissolution medium to the end of the dissolution, the undissolved PAO remained floating on the surface of the dissolution medium, and no suspension was found in the medium, showing 5 poor wettability; and there were relatively fewer floating substances after dissolution experiment. In view of the profile, the API was in a dissolved state ail the way, and the profile did not show slowing down sîgnificantly. In the dissolution process, a large number of impurities were produced, and irregular changes occurred to the impurities. This may be due to the low solubility of phenylarsonic acid, the main dégradation impurity of API, in acid, the 10 dissolution time and state or the like. The related substances substantially tended to be stable
after 24-hour continuons injection of the dissolution sample at 2 h. Light expos tire occurred in the dissolution process, but 24 h stability was measured in the injection plate, which was the stability against light exposure, Therefore, it can be determined that the API was stable in 0.1N hydrochlonc acid. However, strict protection from light w as required in the dissolution 5 process.
[0282] 4.2 Simulated release of PAO in MCT solution and glyceryl behenate solid dispersion
[0283] Sample préparation methods:
[0284] Fl: The same as F15-180929.
(0285] F2: 15 g of glyceryl behenate was heated to 85°C until the glyceryl behenate was melted into a liquid, and then 30 mg of PAO was added and dissolved therein. The resulting product was cooled to room température, and granuiated by sievîng through a 30mesh screen. Thus, a behenate solid dispersion containing 2 mg/g PAO was prepared.
[0286] Experimental methods for simulating release: 5 g of MCT solution and 5 g of 15 the glycerol behenate solid dispersion (2 mg/g) were taken separately (repeated once for each sample). Release experiments were carried out according to the above method with a sampling volume of 3 ml, and no solution was supplemented after sampling. The sample was filtered through a 0.22 pm milîipore filter membrane. The filtrate was investigated for the release by HPLC. After the completion of the dissolution experiment, the floating MCT oil 20 and glyceryl behenate solid dispersion powder were collected, diluted and dissolved, and detected by HPLC to investigate related substances.
]0287] The resuits are shown in FIGs. 2-3 and Tables 42-45:
[0288] Table 42: Cumulative release of MCT solution (Fl)
Sample information MCT sample 1 MCT sample 2
Time (min) Cumulative release (%)
0 0 0
15 15.24 12.81
30 22.78 19.04
45 31.72 17.24
60 39.58 29.96
120 56.29 44.52
Table 43: Cumulative release of glyceryl behenate (F2)
[0289]
Sample information Glyceryl behenate solid dispersion 1 Glyceryl behenate solid dispersion 2
Time (min) Cumulative release (%)
0 0 0
15 1.38 1.28
30 2.99 0.00
45 4.76 2.05
60 7.89 5.78
120 10.97 1.70
[0290] Table 44: Related substances of floating MCT oil after the dissolution expenment of Fl
Name RT % Area % Total impurities
Sample 1 9.271 1.5237 4.13
17.780 95.8681
19.324 0.1311
24.658 0.0494
26.758 0.1878
28.61 0.141
30.37 0.5875
32.566 0.156
34.27 0.0658
36.139 0.1153
39.812 0.7797
40.438 0.3946
[0291] Table 45: Related substances of floating glyceryl behenate after the dissolution experiment of F2
Name RT % Area % Total impurities
Sample 1 5.29 0.023 3.09
9.247 1.5957
17,889 96.9111
19.519 0.0666
24.81 0.085
27.261 0.1406
27.788 0.2861
29.181 0.0558
30.984 0.4433
33.238 0.07
33.985 0.0414
37.368 0.1065
41.045 0.175
Sample 2 5,258 0.0221 2.79
9.115 1.2046
17.773 97,2062
19.426 0.0888
24,705 0,1154
27.152 0.1269
27.662 0.4231
30.841 0.4272
33.086 0.0592
33.716 0.0857
37.076 0.0999
40.8 0.1407
[0292] Analysis: The results are shown in the above tables. Compared with pure APL API was released more slowly from the MCT solution (Fl), and did not reach a dissolution plateau at 2 h. This indicated that PAO was released from the MCT préparation in a sustained 5 manner in the simulated gastric juice, which facilitated to reduce the topical irritation of PAO to the gastric mucosa. PAO was hardly released from the glyceryl behenate solid dispersion (F2). The solid dispersion was prepared from water insoluble glyceryl behenate, and particles were relatively fluffy. Accordîngly, the sample powder was hardly wetted during the dissolution experiments but floated on the surface of the dissolution medium. Therefore, PAO 10 was hardly released. Some impurîties were newly produced for the sample after dissolution experiment. This may be caused by a fa et such as light exposure or by the dissolution medium included in the dissolution residues.
[0293] 4.3 In vitro simulated release experiments of PAO in MC suspension and
PAO in MC suspension with Tween 80
[0294] Sample préparation methods: 10 mg of PAO and 5 g of methylcellulose (MC) aqueous solution (F3, with MC at a concentration of 2%, w/v) or an MC aqueous solution containing 0.1% (w/v) Tween 80 (F4, also with MC at a concentration of 2%. w/v) were weîghed, and magnetically stirred for 30 min. Then. ail the samples were added to the dissolution medium. In addition, samples of the same concentration were prepared respectively, and filtered through a 0.22 pm filter membrane. The content and related substances were detected by HPLC, and comprehensive analysis was performed according to the results.
[0295] Calculation method: The cumulative reiease was calculated by the initially input PAO which excludes PAO dissolved in the initia! suspension. Cumulative reiease = [(API concentration at sampling point * volume of reiease medium) 4- API concentration at previous sampling time point * sampling volume at previous time point]/(input amount - dissolution concentration in suspension * mass of suspension).
[0296]
[0297]
[0298]
The results are shown in FIGs. 4-5 and Tables 46-49:
Table 46; Cumulative reiease of MC suspension
Sample information MC suspension 1 (F3) MC suspension 2 (F4)
Time (min) Cumulative reiease (%)
0 0 0
15 39.25 27.57
30 35.25 27.75
45 48.33 35.51
60 48.99 38.64
120 54.11 33.04
Table 47: Cumulative reiease of MC+0.1% Tween 80 suspension
Sample information MC+ Tween 80 Suspension 1 (F4) MC+ Tween 80 Suspension 2 (F5)
Time (min) Cumulative reiease (%)
0 0 0
15 43.2 24.32
30 50.76 35.66
45 62.16 53.92
60 67.36175 60.16
120 71.32363 63.09175
[0299] Table 48: Détection results of related substances for insoluble substances after the dissolution experiments of MC suspension and MC+Tween 80 suspension
Name RT RRT Area % Area % Total impurities
F3 8.598 0.50 9.9086 0.759 3.07
17.026 1.00 1265.4 96.9253
22.539 1.32 30.23259 2.3157
F4 8.046 0.49 9.33755 2.3907 4.31
16.304 1.00 373.73837 95.6885
21.214 1.30 7.50208 1.9208
[0300] Table 49: Results of related substances for filtrate after sample formulation of
MC suspension and MC+Tween 80 suspension
Name RT RRT Area % Area % Total impurities
F3 8.003 0.49 9.45747 2.2153 2.22
16.275 1.00 417.45660 97.7847
F4 8.046 0.49 9.33755 2.3907 4.31
16.304 1,00 373.73837 95.6885
21.214 1.30 7.50208 1.9208
[0301] Analysis: For the two methylcellulose suspensions, the release substantially reached a plateau at I h, and the cumulative release was close to that of the MCT solution.
However, there were a small number of insoluble substances observed in the actual experiment process. In combination with the results of the related substances in the filtrate after sample formulation in Table 49, it can be inferred that the sample had poor stability in the two media and was highly degraded, thus resulting in a pseudosustained release condition in the simulated release experiments.
[0302] Example 5; In vivo kinetic study of PAO în animais
[0303] 5.1 In vivo kinetic study of oral MCT préparation of PAO in monkeys
[0304] Two groups of monkeys were selected, one male and one femaie in each group.
PAO was orally administered to a first group (male 101 and femaie 102) by taking MCT as a vehicle at a dose of 0.3 mg/kg/day for 2 consecutive weeks. Blood was collected at 0.5, ], 2,
4, 8, 12, 24, and 48 hours after administration on the last day to detect the concentration of the compound in the blood (whole blood, not plasma).
[0305] PAO was orally administered to a second group (male 301 and female 302) by similarly taking MCT as a vehicle at a dose of 0.3 mg/kg via single dosing, Blood was collected at 0.5, 1, 2, 4, 8, 12, 24, and 48 hours after administration to similarly detect the concentration of the compound in the whole blood. Afterwards, the administration was stopped for 5 days before PAO was orally administered by similarly taking MCT as a vehicle at a dose of 0.6 mg/kg via single dosing, Blood was collected at 0.5, 1, 2, 4, 8, 12, 24, and 48 hours after administration to detect the concentration of the compound in the whole blood.
[0306] Table 50: Blood concentrations at different time
Blood sample
_ . Sampling Animal Groupmg m _ , . Average . , Concentration * Analyte , , T . concentration (g/mM (ng/mL)
Compound PAO 0.3 mpk PO 0.5 h 101 102 PI01 (PAO) 80,9 82.2 83.4
301 302 2.71 15.2 27.7
1 h 101 102 93.6 88,3 83.0
301 302 20.9 28,5 36.1
2 h 101 102 109 120 130
301 302 26.6 38,7 50.7
4 h 101 102 172 157 141
301 302 36,7 41.6 46.5
8 h 101 102 26.9 45.1 63.2
301 302 54.7 51.2 47.6
12 h 101 102 27.1 42.4 57.7
301 302 75.9 59.3 42.7
24 h 101 102 89.3 99.7 110
301 302 38.8 33.3 27.8
48 h 101 102 99.8 97.5 95.2
301 302 19,9 21.4 22.8
Compound PAO, 0.6 mpk PO 301 0.5 h 302 31.7 38.6 45.5
301 1 h 302 126 135 144
301 2h 302 116 127 137
301 4 h 302 125 133 141
301 8 h 302 96.6 109 121
301 12 h 302 71.6 85.5 99.4
301 24 h 302 45.5 55.2 64.9
301 48 h 302 32.9 42.0 51.1
[0307] Table 51; pharmacokinetic parameters at 0.3 mpk
Animal ID 101 102 Averag e Animal ID 301 302 Averag e
Rsq_adj ND ND — Rsq_adj ND 0.87 7 —
No. points used for T, 2 0.00 0.00 0.00 Points for T1.2 0.00 5.00 ND
Cmax (ngUïlL) 172 141 157 Cmax (ng/mL) 75,9 50.7 63.3
Tmax (h) 4.00 4.00 4.00 Tmax (h) 12.0 2,00 7.00
T12(h) ND ND ND Tl 2 (h) ND 38.6 ND
Tlast (h) 48.0 48.0 48.0 Thstih) 48.0 48.0 48.0
AUCo-iasi (ng-h/mL) 376 0 450 0 4130 AUCo-ïast (iig-hÎnL) 188 0 1554 1717
AUCo-inf (ng.h/mL) ND ND ND AUCo-inf (ng.h'mL) ND 2823 ND
MRTo-last (h) 26.1 24.8 25,4 MRTo-last (h) 20.8 20.7 20.8
MRTo-inf (h) ND ND ND MRTo-inf (h) ND 58.0 ND
AUCExtm(%) ND ND ND AUCExtra (%) ND 45,0 ND
AUMCExtra (%) ND ND ND AUMCExtra (%) ND 80.4 ND
(0308] Table 52: pharmacokinetic parameters at 0,6 mpk
Animal ID 301 302 Average
Rsq_adj 0.883 0.897 —
No. points used for T1/2 6.00 6.00 6.00
Cmax (ng/mL) 126 144 135
Tmax (h) 1,00 1.00 1.00
T!2(h) 23.5 29.9 26.7
Tlast (h) 48.0 48.0 48.0
AUCo-iast (ng'h/mL) 2807 3796 3302
AUCo-inf (ng.h/mL) 3925 6004 4964
MRTo-iast (h) 18,5 19.7 19.1
MRTo-inf (h) 36,6 46.0 41.3
AUCExtra (%) 28.5 36.8 32.6
AUMCExtra (%) 63.8 72,9 68.4
Notes: ND; Not detemiined (Parameters not determined due to inadequately defined terminal élimination phase)
BQL: Below the lower limit of quantitation (LLOQ)
If the adjusted rsq (linear régression coefficient of the concentration value on the terminal phase) is less than 0.9, T1/2 might not be accurately estimated.
If the % AUCExtra > 20%, AUCo-inf, Cl, MRTo-inf and Vdss might not be accurately estimated.
If the % AUMCExtra > 20%, MRTo-inf and Vd5S might not be accurately estimated.
If the adjusted linear régression coefficient of a final phase concentration value is less than 0.9, then T1-2 probably cannot be accurately estimated.
[0309] Analysis: After a single oral administration of the MCT préparation of PAO to the monkeys, PAO can be absorbed into the blood, and the blood concentration reached the maximum within 4 hours. PAO had a long half life in the blood, being about 26.7 hours, which indicated that a higher blood concentration can be maintained by orally administration of PAO once a day. In conclusion, this indicated that the MCT préparation of PAO can be used for oral delivery of PAO. After the MCT préparation of PAO was orally administered to the monkeys every day at a dose of 0.3 mg/kg/day for 2 consecutive weeks, the average exposure of PAO in the blood (AUCo.jaÎ1 = 4130 ng'h/mL) was about 2.4 times as high as that of a single oral administration (AUCo-iast =1717 ng*h/mL), indicating that repeated administration will cause médicament accumulation. It was suggested that the administration should be stopped for 1-2 days after the consecutive daily oral administration of the médicament for 2 weeks or within 2 weeks.
[0310] 5.2 In vivo kinetic study of oral sesame oil préparation of PAO and intravenous PAO in monkeys
[0311] Fatty acids in MCT are mainly medium-chain saturated fatty acids, while fatty acids in sesame oil are mainly long-chain unsaturated fatty acids. There are significant différences between the two. Also, long-chain fatty acids are mainly absorbed by lymphatic vessels în the intestine, while medium-chain fatty acids are mainly absorbed by intestinal mucosal cells. Therefore, we detected the kinetics of a sesame oil préparation of PAO orally administered to the monkeys and compared them with the kinetics of intravenous PAO.
[0312] Two groups of monkeys were selected, ail of which were male. PAO was administered to a first group (C 1001 and Cl002) through iv injection by taking 1% DMSO as a vehicle at an actual dose of 0.118 mg/kg (nominal dose; 0.100 mg/kg) via single dosing. PAO was orally administered to a second group (C2001 and C2002) by taking sesame oil as a vehicle at an actual dose of 0.168 mg/kg (nominal dose: 0.200 mg/kg) via similariy single dosing. For each group, blood was collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, 24, and 48 hours after administration to detect the concentration ofthe compound in the blood (whole blood, not plasma).
[0313] The results are shown in Table 53 and FIGs. 6A-6C:
[0314] Table 53; Results of in vivo kinetic study in monkeys
Pharmacokinetics in monkeys (ng/mL)
PAO
PO
IV time (h) C1001 Cl 002 Average IV PO time (h) C2001 C2002 Average PO
0.0000 BQL BQL ND 0.000 BQL BQL ND
0.0830 2400 2470 2435 0.0830 BQL BQL ND
0.250 . 1040 1230 1135 0.250 4.11 20.4 12.3
0.500 513 678 596 0.500 14.0 45.0 29.5
1.00 236 256 246 1.00 36.0 41.4 38.7
2.00 112 113 113 2.00 35.1 45._3 40.2
4.00 . 68.7 87.6 78.2 4.00 41.4 43.5 42.5
8.00 40.5 47.1 43.8 8.00 25.1 26.0 25.6
12.0 36.6 39.6 38.1 12.0 21.6 19.4 20.5
24.0 21.8 26.9 24.4 24.0 13.8 11.9 12.9
48.0 23.2 20.8 22.0 48.0 8.88 9.00 8.94
Rsq_adj 0,516 0.869 — Rsq_adj 0.949 0.812 —
No. points used for 6.00 4.00 ND No. points used for 4.00 4,00 4.00
Tu2 Ti?
Co (ngiiiL) 3637 3493 3565 Cn™ (ng/mL) 41.4 45.3 43.4
Tl? (11) 24.3 35.3 29.8 Tma.x (h) 4.00 2.00 3.00
Vdss (L/kg) 0.894 0.933 0.914 T 1.2 (h) 27.0 28.0 27.5
Cl (mL/minkg) 0.507 0.436 0.471 (h) 48.0 48.0 48.0
T|asi (h) 48.0 48.0 48.0 AUCo-iasi (ng'lVmL) 828 824 826
AUCo-i^t (ng.h/mL) 2477 2762 2620 AUCo-inf (ng.h/mL) 1174 1187 1181
AUCo-inf (ng.h/mL) 3289 3821 3555 MRTo-last (h) 18.2 17.1 17.7
MRTo-last (h) 11.8 11.4 11,6 MRTo-mf (h) 38.5 38.9 38.7
MRTo-mf (h) 29.4 35.7 32.5 AUCExtra (%) 29.5 30.6 30.0
AUCExtra(%) 24.7 27.7 26.2 AUMCExtra (%) 66.7 69.5 68.1
AUMCExtra (%) 69.7 76.9 73.3 Bioavailability (%)a 15.8
Notes: ND: Not determined (Parameters not determined due to inadequately defined terminal élimination phase)
BQL: Below the lower limit of quantitation (LLOQ)
If the adjusted rsq (iinear régression coefficient of the concentration value on the terminal phase) is less than 0.9, Ti,2 might not be accurately estimated.
If the % AUCExsra > 20%, AUCo.inf, Cl, MRTo-mf and Vdss might not be accurately estimated.
If the % AUMCExtra > 20%, MRT0-mf and Vdss might not be accurately estimated.
If the adjusted Iinear régression coefficient of the concentration value on the terminal phase is less than 0.9, T1/2 might not be accurately estimated.
a: Bioavailability (%) was calculated using AUCo-mf (% AUCexî™ < 20%) or AUCo-iast (% AUCExtra > 20%) with nominal dose.
]0315] Analysis: After a single oral administration of the sesame oil préparation of PAO to the monkeys (at a dose of 0.168 mg/kg), PAO can also be absorbed inio the blood, and the blood concentration similariy reached the maximum within 4 hours. The half life of PAO in the blood was about 27.5 hours. The average exposure of PAO in the blood (AUCo-ia« = 826 ngh/mL) was about 0.48 times as high as the average exposure of a single oral administration ofthe MCT préparation of PAO at 0.3 mg/kg (AUCo-iast =1717 ng hmL) and about 0.25 times as high as the average exposure at 0.6 mg/kg (AUCo-um = 3302 ng’hTnL). This indicated that the oral sesame oil préparation and MCT préparation of PAO were comparable in in vivo kinetics and bioavailability.
[0316] 5.3 In vivo kinetic study of oral MCT préparation of PAO in beagles
[0317] Two groups of beagles were selected, ail of which were male. PAO was administered to a first group (DI001 and DI002) through iv injection by taking 1% DMSO as a vehicle at an actual dose of 0.101 mg/kg (nominal dose: 0.100 mg/kg) via single dosing. PAO was orally administered to a second group (D2001 and D2002) by taking sesame oil as a vehicle at an actual dose of 0.169 mg/kg (nominal dose: 0.200 mg/kg) via similariy single dosing. For each group, blood was collected at 0.083, 0.25, 0.5, 1,2, 4, 8, 12, 24, and 48 hours after administration to detect the concentration of the compound in the blood (whole blood, not plasma).
[0318] The results are shown in Table 54 and FIG s. 7A-7C:
[0319] Table 54: Results of in vivo kinetic study in beagles
Pharmacokînetîcs in beagles (ng/mL)
PAO
IV PO
IV time (h) D1001 DI 002 Average IV PO time (h) D2001 D2002 Average PO
0.0000 BQL BQL ND 0.000 BQL BQL ND
0.0830 603 657 630 0.0830 BQL BQL ND
0.250 238 411 325 0.250 6.36 BQL ND
0.500 146 219 183 0.500 14.5 6.72 10.6
1.00 68.4 102 85.2 1.00 18.1 10.9 14.5
2.00 43.2 75.0 59.1 2.00 20.6 16.7 18.7
4.00 20.6 31.5 26.1 4.00 16.1 15.5 15.8
8.00 13.5 17.4 15.5 8.00 9.81 11.3 10.6
12.0 7.65 12.9 10.3 12.0 7.62 10.6 9.11
24.0 6.27 8.49 7.38 24.0 4.02 5.88 4.95
48.0 4.41 5.46 4.94 48.0 BQL BQL ND
Rsq_adj 0.998 0.940 — Rsq_adj 0.997 0.967 —
No. points used for T] 7 3.00 3.00 3.00 No. points used for Tl2 3.00 4.00 ND
Co (ng/mL) 957 830 893 Cmax (ng/mL) 20.6 16.7 18.7
T12(h) 45.6 30,0 37.8 TmaxCh) 2.00 2.00 2.00
VdSÎ (L/kg) 4.42 2.27 3.35 Tii(h) 12.6 15.0 13.8
Cl (mL/min/kg) 1.75 i.42 1.59 Tlast (h) 24.0 24.0 24.0
Tïasi (h) 48.0 48.0 48.0 AUCo-iast (ng-h/mL) 220 245 233
AUCo.w (ng.h'mL) 662 936 799 AUCo-inf (ng.h'mL) 293 372 333
AUCo-inf (ng.h'mL} 952 1173 1062 NlRTo-iaït (h) 8,89 10.4 9.64
MRTo-last (11) 10.7 10.3 10.5 MRTo-inf (h) 17.1 22.4 19.8
MRTo-mf (h) 42.1 26.6 34.4 AUCExtra (%) 24.8 34.1 29.5
AUCexM (%) 30.5 20.2 25.3 AUMCExtra (%) 61.0 69.5 65.3
AUMCExtra (%) 82.3 69.1 75.7 Bioavaîlability (%)a — - 14.6
Notes: ND: Not determined (Parameters not determined due to inadequately defined terminal élimination phase)
BQL: Below the lower lirait of quanti talion (LLOQ)
If the adjusted rsq (linear régression coefficient of the concentration value on the terminal phase) is less than 0.9, T 1,2 might not be accurately estimated.
If the % AUCExtra > 20%, AUCo-mf, Cl, MRTo-inf and Vdi5 might not be accurately estimated.
If the % AUMCExtra > 20%, MRTo-inf and Vdss might not be accurately estimated.
If the adjusted linear régression coefficient of the concentration value on the terminal phase is less than 0.9, T1/2 might not be accurately estimated, a: Bioavaîlability (%) was calculated using AUCo-inf (% AUCExtra < 20%) or AUCo-iaSi (% AUCExtra > 20%) with nominal dose.
[0320] Analysis: After a single oral administration of the sesame oil préparation of
PAO to the beagles, the blood concentration also reached the maximum within 4 hours, and the bioavaîlability was similar, being about 15%. However, the exposure of PAO in the blood of the beagles was about one fourth of the exposure in the blood of the monkeys, This indicated that the lipid préparation of PAO has similar bioavaîlability in different animal species, but has relatively large différence in exposure.
[0321 ] 5.4 In vivo kinetic study of oral DMSO préparation and MCT préparation of PAO in mice
[0322[ In order to compare the différence between the oral DMSO préparation and the oral MCT préparation, in vivo kinetic study in mice was carried out. Maie mice were
dwided into two groups, 3 mice per group. PAO was oraily administered to one group (MOI, M02 and M03) by taking a 1% DMSO aqueous solution as a vehicle at an actual dose of 0.0913 mg'kg (nominal dose: 0.100 mg/kg). The MCT préparation of PAO was administered to the other group (N01, N02 and N03) at an actual dose of 0.107 mg/kg (nominal dose:
0.100 mg'kg). Blood was collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours after administration to detect the concentration of the compound in the blood (whole blood, not plasma).
[0323] The results are shown in Tables 55-56 and FIGs. 8A-8B;
[0324] Table 55: Results of in vivo kinetic study of oral 1% DMSO préparation of
PAO (dose: 0.0913 mg/kg) in mice
Pharmacokinetics of PAO in mice (ng/mL)
PAO
PO
PO time (h) MOI M02 M03 Average PO SD CV (%)
0.0830 1.39 0.678 0.783 0.950 i 0.384 40.4
0.250 1.82 0.916 1.08 1.27 0.482 37,9
0.500 2.02 0.899 0.951 1.29 0.633 49,0
1.00 2.27 0.874 1.22 1.45 i 0,727 50.0
2.00 1.27 1.04 1.29 1,20 St 0.139 11,6
4.00 2.59 1.29 1.38 1.75 i 0.726 41.4
6.00 1.89 1.30 1.21 1.47 0.369 25.2
8.00 1.69 1.07 0.954 1.24 i 0.396 32.0
24.0 0.509 BQL BQL ND ND ND
PK parameters MOI M02 M03 Average PO SD CV (%)
Rsq_adj 0.999 ND ND — d: —
No. points used for Tlz 3.00 0,00 0.00 ND zt — —
(ng/mL) 2.59 1.30 1.38 1,76 ± 0.723 41.1
1^(11) 4.00 6.00 4.00 4.67 dz 1.15 24.7
T i,, (h) 9.41 ND ND ND ± ND ND
T|ast (11) 24.0 8.00 8.00 ND ± — —
AUCo-insi (ng'h/mL) 31.3 9.07 9.65 16.7 ± 12.7 75.9
AUCo-inf (ng.h/mL) 38.2 ND ND ND ± ND ND
MRTo-last (h) 9.29 4.25 3.98 5.84 ± 2.99 51.2
MRTo-uif (h) 14.4 ND ND ND ± ND ND
AUCExi™(%) 18.1 ND ND ND ± ND ND
AUMCEma(%) 47.2 ND ND ND i ND ND
Notes: ND: Not determined (Parameters not determined due to inadequately defined terminal élimination phase)
BQL: Below the lower limit of quantitation (LLOQ)
If the adjusted rsq (linear régression coefficient of the concentration value on the terminal phase) is less than 0.9, T12 might not be accuraîely estimated.
If the % AUCrxtra > 20%, AUCo-w, Cl, MRTo-inf and Vd« might not be accurately estimated.
If the % AUMCexiw > 20%, MRTo-inf and Vdss might not be accurately estimated.
If the adjusted linear régression coefficient of the concentration value on the terminal phase is less than 0.9, T1-2 might not be accurately estimated.
[0325] Table 56: Results of in vivo kinetic study of oral MCT préparation of PAO (dose: 0.107 mg/kg) in mice
Pharmacokinetics of PAO in mice (ng/mL)
PAO
PO
PO time (h) N01 N02 N03 Average PO SD CV (%)
0.0830 BQL BQL BQL ND ± ND ND
0.250 1.06 2.78 1.42 1.75 0.907 51.7
0.500 0.875 4.57 1.95 2.47 ά: 1.90 77.1
1.00 BQL 5.47 1.80 3-64 ND ND
2.00 BQL 2.81 1.10 1.96 ± ND ND
4.00 0.714 2.07 0.926 1.24 ± 0.729 59.0
6.00 0.916 2.47 1.12 1.50 ± 0.844 56.2
8.00 0.857 2.12 1.85 1.61 ± 0.665 41.3
24.0 BQL 0.822 1.57 1.20 ± ND ND
PK parameters N01 N02 N03 Average PO SD CV (%)
Rsq_adj ND 0.999 ND — — —
No. points used for T1/2 0.00 3.00 0.00 ND ± — —
Cœax (ng/mL) 1.06 5.47 1.95 2.83 2.33 82.5
Tmax (h) 0.250 1.00 0.500 0.583 ± 0.382 65.5
T1/2 (h) ND 11.5 ND ND ND ND
T last (11) 8.00 24.0 24.0 ND —
AUCo-iasi (ngh/rnL) 6.54 43.6 37.2 29.1 ± 19.8 68.1
AUCo-mf (ng.h/mL) ND 57.3 ND ND ± ND ND
MRTû-last (h) 4.09 9.18 12.6 8.64 4.30 49.8
MRTo-inf (h) ND 16.6 ND ND ND ND
AUCExtta(%) ND 23.8 ND ND ± ND ND
AUMCexm (%) ND 57.9 ND ND ND ND
Notes: ND: Not determined (Parameters not determined due to inadequately defined terminal élimination phase)
BQL: Below the lower limit of quantitation (LLOQ)
If the adjusted rsq (linear régression coefficient of the concentration value on the terminal phase) is less than 0.9, Ti/2 might not be accurately estimated.
If the % AUCExtn > 20%, AUCo-inf, Cl, MRTo-îbî and Vdss might not be accurately estimated.
If the % AUMCExtra > 20%, MRTo-mf and Vd&s might not be accurately estimated.
If the adjusted linear régression coefficient of the concentration value on the terminal phase is less than 0.9, Ti-2 might not be accurately estimated.
[0326] The results showed that the bioavailability of PAO delivered by the MCT préparation was apparently higher than the bioavailability of PAO delivered by the cosolvent DMSO aqueous solution. In addition, Example 4.2 and Table 42 of the présent invention showed that the PAO was released from its MCT préparation in a sustained manner in the simulated gastric juice. Therefore, the lipid préparation of PAO can not only realtze the sustained release of PAO in the gastric juice so as to relieve irritation of PAO to the gastric mucosa, but also increase the bioavailability of PAO,
[0327] 5,5 In vivo kinetic study of PAO in rats
[0328] In order to compare the différences between PAO préparations in administration route, dose and gender of administered subjects, in vivo kinetic study in rats was carried out. Rats were divided into four groups, 6 rats per group. Each group included 3 female rats and 3 male rats. PAO was ùitravenously administered to a first group at a nominal dose of 0.1 mg/kg. PAO was orally administered to a second group at a nominal dose of 0.1 mg/kg. PAO was orally administered to a third group at a nominal dose of 0.3 mg/kg. PAO was orally administered to a fourth group at a nominal dose of 0.9 mg/kg. The concentration of the compound in the blood (whole blood, not plasma) within 36 hours after administration was detected.
[0329] The results are shown in Table 57:
[03301 Table 57: Average pharmacokinetic parameters of PAO in male and female SD rats (n=6)
Group 1 2 3 4
Administration route IV PO PO PO
Dose level (mg/kg) 0.1 0.1 0.3 0.9
PK parameters Average SD Average SD Average SD Average SD
Co or Cmai (ng/mL) 1390 130 71.7 30.2 262 55.1 665 174
Tmax (h) — — 4.67 1.03 5.33 2.07 7.00 3.03
TU (h) 7.59 0.992 7.26 1.11 6.83 0.769 7.68 1.59
Vdss(L/kg) 0.130 0.011 4 — — — — —
CL (mL/min/kg) 0.343 0.031 2 — — — —
AUCo-bst (h*ng/mL) 4790 432 739 288 2950 694 9120 2060
AUCo-iaf (h*ng/mL) 4890 436 767 294 3050 700 9710 1960
Bioavailability (%) — — 15.7 — 20.8 — 22.1 —
a: Bioavaîlability (%) was calculated with average AUCo-mf and nominal dose. means not applicable
[0331] Table 58: Dose ratio of PAO in male and female SD rats after single oral administration of PAO
Gender Compared Doses (mg/kg) Dose ratio Cmai value Cmai ratio AUCo-last value AUCoratio
Male 0.300 over 0.100 3.00 230/59.6 3.86 2560/666 3.84
0.900 over 0.300 3.00 580/230 2.52 8270/2560 3.23
0.900 over 0.100 9.00 580/59.6 9.73 8270/666 12.4
Female 0.300 over 0.100 3.00 294/83.8 3.51 3330/812 4.10
0.900 over 0.300 3.00 749/294 2.55 9970/3330 2.99
0.900 over 0.100 9.00 749/83.8 8.94 9970/812 12.3
[0332] Table 59; Gender comparison of PAO on systemîc exposure in male and female SD rats after single intravenous or oral administration of PAO
Administration route Dose level (mg/kg) Cfl ΟΓ Cniax (female and male) Cf) ΟΓ Cmax ratio (female/male) AUCo-last value (female and male) AUCo-last ratio (female/male)
IV 0.100 1420/1350 1.05 4630/4940 0.937
PO 0.100 83.8/59.6 1.41 812/666 1.22
PO 0.300 294/230 1.28 3330/2560 1.30
PO 0.900 749/580 L29 9970/8270 1.21
[0333] 5.6 In vivo kinetic study of PAO in rats (0334] In order to study the influences of éthanol on the oral MCT préparation of PAO, in vivo kinetic study in rats was carried out. Rats were divided into two groups, 2 rats per group, one male and one female in each group. MCT préparations of PAO (PAO in MCT solution, having a concentration of 1.5 mg/g) were orally administered to one group (ROI and 15 R02) by using a vehicle contaîning no éthanol at a dose of 0.1 mpk. MCT préparations of
PAO (contaîning 1.05% (v/v) of éthanol) were orally administered to the other group (R01 and R02) at a dose of 0,2 mpk, Blood was collected at 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours after administration to detect the concentration of the compound in the blood.
[0335] The results are shown in Table 60:
[0336] Table 60: Influences of éthanol on the PK of MCT préparations of PAO orally 5 administered to SD rats
Vehicle (containing no éthanol) DMPK formulation (containing 1.05% (v/v) of éthanol)
At a dose ofO.l mpk At a dose of 0.2 mpk
POi (male) PO2 (female) POI (male) PO2 (female)
PO time (h) ROI R02 ROI R02
0.250 3,73 4.35 14.4 20.0
0.500 8.89 7.93 30.2 49.2
1.00 11.7 12.0 47.1 64,8
2.00 17.4 11.2 61.4 117
4.00 17.8 14.0 98.3 210
6.00 14.4 13.7 121 193
8.00 10.6 11.9 96.2 140
24.0 6.58 3.69 11.0 15,4
PK parameters ROI R02 ROI R02
Rsq_adj 0.829 1,00 1.00 1.00
No. points used for Tp2 3,00 3.00 3.00 3.00
Cmax (ng/mL) 17.8 14.0 121 210
Tmax (11) 4.00 4.00 6.00 4.00
Tl,2 (h) 18.1 9.50 5.17 4.97
Tlast(h) 24.0 24.0 24.0 24.0
AUCo-iast (ng’h/mL) 249 209 1305 2094
AUCo-inf (ng.h/mL) 421 260 1387 2204
MRTo-iast (h) 10.2 9.75 8.90 8.3S
MRTo-mf(h) 26.5 15.2 10.2 9.52
AUCextra (%) 40.9 19.5 5.91 5.01
AUMCExtra(%) 77.3 48.3 18.2 16.4
[0337] Table 61: Summary of influences of éthanol on the PK of MCT préparations of PAO orally administered to SD rats
Préparation Dose AUCo-lasi Average Compared with the préparation containing no éthanol at the same dose
Containing no éthanol 0.1 249 209 229
Containing éthanol 0.2 1305 1700 3.71
2094
[0338] It can be seen that adding éthanol to the MCT préparation of PAO can increase 5 the exposure of oral PAO in the blood or increase the bioavailability of oral PAO. When the concentration of the éthanol was 1.05% (wv), the bioavailability can be increased by 2-3 times.
[0339] Example 6: Toxicity study of different PAO préparations in animais [0340] In order to compare the toxicity of an MCT préparation of PAO and a 0.1%
DMSO aqueous solution of PAO in animais, 20 male ICR mice and 20 female ICR mice were selected, ail of which were 10 weeks old. The male and female mices were equally divided into 4 groups to which the MCT préparation of PAO and the 0.1% DMSO aqueous solution (v/v) of PAO were intragastrically administered at 1.5 or 0.75 mg/kg/day respectively for 46 days (Conditions for grouping and dosing of mice are shown in Table 62).
The mice were weighed every day, and dead mice were documented.
[0341] Table 62; Grouping and dosing of mice
Préparation Dose Number of female mice Number of male mice
MCT préparation 1.5 mg/kg/day 5 5
0.75 mg/kg/day 5 5
0.1% DMSO aqueous solution 1.5 mg/kg/day 5 5
0.75 mg/kg/day 5 5
[0342] After consecutive administration for 46 days, ail the female mice survived.
The average weights of the two groups of female mice to which the MCT préparation and the 20 0.1 % DMSO aqueous solution of PAO were orally administered at 0.75 mg/kg/day increased slowly, and there was almost no différence between the two groups when the administration was completed (30.6 g and 30.2 g). The average weights of the female mice to which the
MCT préparation of PAO was orally administered at 1.5 mg/kg/day slowly increased to 32.9 g. However, the average weights of the mice to which the 0.1% DMSO aqueous solution of PAO was orally administered decreased significantly after the second week, and finally dropped to 24.4 g (FIG. 9). As for the male mice, every mouse to which the MCT préparation 5 of PAO was orally administered at 0.75 mg/kg/day sun ived, and the weight of each mouse increased slowly. However, there was no obvions regularity for the weight changes of the mice in the other three groups. The spécifie results were as follows: one of the 5 mice to which the MCT préparation of PAO was orally administered at 1.5 mg/kg/day died in the second week of administration; one of the 5 mice to which the 0.1% DMSO aqueous solution 10 of PAO was orally administered at 0.75 mg/kg/day died in the second week of administration;
and two ofthe 5 mice to which the 0.1% DMSO aqueous solution of PAO was orally administered at 1.5 mg/kg/day died in the second and sixth week of administration, respectively.
[0343] These results showed that although the bioavailability of PAO delivered by the 15 MCT préparation was apparently higher than the bioavailability of PAO delivered by the aqueous solution of cosoivent DMSO, the in vivo toxicity of the MCT préparation of PAO was significantly lower.

Claims (42)

1. A pharmaceutical composition, comprising a micromoiecule PI4KIIIa inhibitor and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier comprises a lipid.
2. The pharmaceutical composition according to claim 1, wherein the micromoiecule PI4KIIIa inhibitor is PAO and a dérivative of PAO.
3. The pharmaceutical composition according to claim 1, wherein the micromoiecule PMKIIIa inhibitor has a structure of formula (I) or a pharmaceutically acceptable sait thereof, ^(Ri)n formula (I) wherein Ri is each independently selected from H, halogen, nitro, cyano, hydroxyl, amino, carbamoyl, Ci-6 alkyl, Ci-6 alkoxy, Ci-6 haloalkyl, -As(O), -NH-(Ci-6 alkyl), N,N-(Ci-6 alkyl)2, -NH-C(O)-R2, -NH-S(O)2-R3, -C(O)OR4 or heterocyclyl, wherein n is an integer of 0-5, R2 and Rs are each independently selected from H, amino, Ci-6 alkyl, Ci-6 alkoxy, Ci-6 haloalkyl, -NH-(Ci-6 alkyl), N,N-(Ci.& alkyl)2, C(O)OR4, C3-6 cycloalkyl, 6-12 membered aryl or 3-6 membered heterocyclyl, which are optionally substîtuted by halogen, nitro, cyano, hydroxyl, amino, carbamoyl, aryl, C1-6 alkyl, C2-6 alkynyl, C2-6 alkenyl, C].f> alkoxy, Ci-6 haloalkyl, 3-6 membered heterocyclyl, C3-6 cycloalkyl or Bn-O-, and R4 is C1-6 alkyl.
4. The pharmaceutical composition according to claim 3, wherein Ri is each independently selected from H, halogen, nitro, cyano, hydroxyl, amino, carbamoyl, Ci-6 alkyl, C1.6 alkoxy, Ci-6 haloalkyl, -As(O), -NH-(Cp6 alkyl), N,N-(Ci-6 alkyi)2 or -C(0)0R4, wherein n is an integer of 0-2, and R4 is C1-6 alkyl.
5. The pharmaceutical composition according to ciaîm 3, wherein Ri is each independently selected from H, halogen, nitro, cyano, hydroxyl, amino, C1-6 alkyl, Ci-6 alkoxy, Ci-6 haloalkyl or -As(O), wherein n is an integer of 0-2.
6. The pharmaceutical composition according to claim 3, wherein Ri is each independently selected from H, halogen, amino or Ci-6 alkoxy, wherein n is 1.
7. The pharmaceutical composition according to claim 6, wherein Rj is located at an ortho position or a para position of the -As(O) group.
8. The pharmaceutical composition according to claim 3, wherein Ri is H.
9. The pharmaceutical composition according to claim 1, wherein the micromolecule
PI4KIIIa inhibitor is at an amount of 0.01-20 mg/g, 0.05-20 mg/g, 0.1-20 mg/g, 0.2-20 mg/g, 0.5-20 mg/g, 0.8-20 mg/g, 1-20 mg/g, 1-18 mg/g, 1-16 mg/g, 1-14 mg/g, 1-12 mg/g, 1-10 mg/g, 2-10 mg/g, 2-8 mg/g, 2-6 mg/g, 3-6 mg/g, 0.2-15 mg/g, 0.2-12 mg/g, 0.2-10 mg/g, 0.28 mg/g, 0.2-6 mg/g, 0.2-4 mg/g, 0.2-2 mg/g, 0.2-1 mg/g or 0.2-0.8 mg/g in the pharmaceutical composition.
10. The pharmaceutical composition according to claim 1, wherein tire pharmaceutically acceptable carrier comprises at least about 50% (w/w), at least about 60% (w/w), at least about 70% (w/w), at least about 80% (w/w), at least about 85% (w/w), at least about 90% (w/w), at least about 95% (w/w), at least about 97% (w/w), at least about 98% (w/w), at Ieast about 99% (w/w) or 100% (w/w) of the lipid.
11. The pharmaceutical composition according to any one of the preceding daims, wherein the lipid comprises a lipid with a melting point of -20-80°C, -20-10°C or -20-0°C.
12. The pharmaceutical composition according to any one of the preceding daims, wherein the lipid has a degree of unsaturatîon of 0-5, 0-4, 0-3, 0-2, 0-1 or 0.
13. The pharmaceutical composition according to any one of the preceding daims, wherein the lipid comprises a lipid which has a fatty acid carbon chain at a length in a range of 4-24, 4-22, 4-20, 6-20, 6-16, 6-14, 6-13, 6-12, 8-13, 8-12 or 8-10 carbon atoms.
14. The pharmaceutical composition according to any one of the preceding daims, wherein the lipid comprises a lipid which has a fatty acid chain at a length of 8 and 10, and optionally further comprises a lipid which has the fatty acid carbon chain at a length of 12-22.
15. The pharmaceutical composition according to any one of the preceding daims, wherein the fatty acid chain in the lipid is a long-chain fatty acid, a medium-chain fatty acid or a short-chain fatty acid.
16. The composition according to claim 1, wherein the lipid is vegetable oil.
17. The pharmaceutical composition according to claim 16, wherein the vegetable oil îs olive oil, tea oil, rapeseed oil, peanut οίl, soybean oil, corn oil, safflower oil, groundnut oil, sunflower seed oil, canola oil, walnut oil, almond oil, avocado oil, castor oil, coconut oil, cottonseed oil, rice bran oil, sesame oil, refmed palm oil or a mixture thereof.
18. The pharmaceutical composition according to any one of the preceding daims, wherein the lipid is a fatty acid, a fatty acid ester, a fatty alcohoL a lipoîd, a paraffm or a mixture thereof.
19. The pharmaceutical composition according to claim 18, wherein the lipoid is a phospholipîd, a sucrose ester, a steroid, a fat-soluble vitamin or a mixture thereof.
20. The pharmaceutical composition according to claim 18, wherein the fatty acid ester is a glyceride, an ethylene glycol ester, a propylene glycol ester or a mixture thereof.
21. The pharmaceutical composition according to claim 18, wherein the fatty acid ester is a monoester, a diester, a triester or a mixture thereof.
22. The pharmaceutical composition according to claim 18, wherein the fatty acid ester comprises glycerides of octanoic acid and/or decanoic acid.
23. The pharmaceutical composition according to daim 18, wherein the fatty acid ester comprises a medium-chain triglycéride.
24. The pharmaceutical composition according to any one of the preceding daims, wherein the pharmaceutically acceptable carrier further comprises an antioxidant.
25. The pharmaceutical composition according to claim 24, wherein the antioxidant is at an amount ofû.001%-5% (wt), O.OO5%-5% (wt), 0.01%-5% (wt), 0.05%-5% (wt), 0.1%-5% (wt), 0.1%-3% (wt), Û.1%-2% (wt), 0.1%-l% (wt), 0.1%-0.8% (wt), 0.1%-0.5% (wt), 0.1%0.3% (wt), 0.3%-2% (wt), 0.5%-2% (wt), 0.8%-2% (wt) or l%-2% (wt) based on the weight of the pharmaceutical composition.
26. The pharmaceutical composition according to claim 24, wherein the antioxidant is sulfite, bisulfite, pyrosulfite, dithiocarbamate, ascorbîc acid, ascorbyl palmitate, hydrocoumarin, vitamin E, ethanolamine, propyl gallate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), nordihydroguaiaretic acid or glutathione.
27. The pharmaceutical composition according to any one of the preceding daims,
wherein the pharmaceutically acceptable carrier further comprises a viscosity modifier, a pH regulator or a flavoring agent.
28. The pharmaceutical composition according to any one of the preceding claims, wherein the pharmaceutically acceptable carrier further comprises éthanol.
29. The pharmaceutical composition according to claim 28, wherein the éthanol is at an amount of 1ü%-0.1 % (v/v).
30. The pharmaceutical composition according to claim 28, wherein the éthanol is at an amount of 8%-0.1% (v/v), 7%-0.1% (v/v), 6%-0.1% (v/v), 5%-0.1% (v/v), 4%-0.1% (v/v), 3%-0.1% (v/v), 2%-0.1% (v/v), 1,5%-0.1% (v/v), 1.2%-0.1% (v/v), 8%-0.3%(v/v), 8%0.5%(v/v), 8%-0.7%(v/v), 8%-0.9% (v/v), 8%-l% (v/v), 6%-0.3% (v/v), 5%-0.5% (v/v), 4%0.8% (v/v), 3%-0.9% (v/v) or 2%-l% (v/v).
31. The pharmaceutical composition according to any one of the preceding claims, wherein the pharmaceutical composition is used for oral, subcutaneous, intrainuscular or intravenous administration.
32. The pharmaceutical composition according to any one of the preceding claims, wherein the pharmaceutical composition îs tablets, capsules, suspensions, émulsions, solutions, semisolid préparations, patches or microneedles.
33. The pharmaceutical composition according to claim 1, wherein the micromoiecule ΡΙ4ΚΠΙα inhibitor is phenylarsine oxide, the phenylarsine oxide is at an amount of 0.25-20 mg/g in the pharmaceutical composition, and the pharmaceutically acceptable carrier is consisting of a medium-chain triglycéride, consisting of a medium-chain triglycéride and a long-chain triglycéride, or consisting of a medium-chain triglycéride and éthanol.
34. A method for preparing the pharmaceutical composition according to any one of claims 1-33, wherein the method comprises: mixing the micromoiecule Ρ14ΚΙΠα inhibitor and the pharmaceutically acceptable carrier to obtain a mixture.
35. The method according to claim 34, wherein the method comprises: mixing the micromoiecule PMKIlIa inhibitor and the pharmaceutically acceptable carrier through a mechanical force.
36. The method according to claim 35, wherein the mechanical force is stirring,
dispersing. shaking or ultrasonic treatment.
37. The method according to claim 34, wherein the method comprises: mixing the micromolecule PI4KIIIa inhibitor and the pharmaceuticaliy acceptable carrier after melting the pharmaceuticaliy acceptable carrier by heating.
5
38. The method according to claim 37, further comprising: filtering the mixture.
39. Use of the pharmaceutical composition according to any one of claims 1-33 in the manufacture of a médicament for treating a PI4KTIIa-related disease in a subject.
40. The pharmaceutical composition according to any one of claims 1 -33 for use in treating a PI4KIlIa-related disease in a subject.
10
41. The use according to claim 39 or the pharmaceutical composition for use according to claim 40, wherein the PMKlIIa-related disease is Alzheimer's disease.
42. The use according to claim 39 or the pharmaceutical composition for use according to claim 40, wherein the subject is an animal such as a pig, a dog, a monkey, a cat, a mouse, or a rat, or a human.
OA1202200005 2019-07-05 2020-07-06 Micromolecule PI4Klllα inhibitor composition, preparation method therefor and use thereof. OA20568A (en)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CNPCT/CN2019/094831 2019-07-05

Publications (1)

Publication Number Publication Date
OA20568A true OA20568A (en) 2022-10-27

Family

ID=

Similar Documents

Publication Publication Date Title
CA2866067C (en) Uses of certain f-series prostaglandin analogs for treating diabetes and dyslipidemia
TW416849B (en) New pharmaceutical composition based on taxoids
RU2393160C2 (en) Pharmaceutical composition containing temozolomide ester
US20230115711A1 (en) MICROMOLECULE PI4KIIIalpha INHIBITOR COMPOSITION, PREPARATION METHOD THEREFOR AND USE THEREOF
CN102344408B (en) Double-effect anesthetic
CN113336768B (en) Multi-target tyrosine kinase inhibitor
WO2015110040A1 (en) Chlorambucil derivative, preparation method and application
OA20568A (en) Micromolecule PI4Klllα inhibitor composition, preparation method therefor and use thereof.
EP0561597B1 (en) New derivatives of physostigmine, their use and pharmaceutical formulations containing them
CN111166745A (en) Composition containing racemic oxypyramine or salt thereof and application
CN114728883B (en) Ibuprofen ester prodrug, pharmaceutical composition, preparation method and application
JP3277735B2 (en) Composition for promoting absorption of naphthoic acid derivative
CN101456814A (en) Dexibuprofen guaiacol ester and preparation method thereof
SK284247B6 (en) Liposome formulation of 6,9-bis-[(2-aminoethyl)- amino]benzo[g]isoquinoline-5,10-dione dimaleate
EP1648413B1 (en) Pharmaceutical compositions of short-acting sedative hypnotic agent
JP7571940B2 (en) Ibuprofen ester derivatives and emulsion formulations thereof
KR20150030725A (en) Anti lung cancer drug made of oleum cassiae and major constituent cinnamaldehyde thereof and use thereof
CN115531306B (en) Left ornidazole derivative emulsion preparation for injection and preparation method thereof
CN109985004A (en) Flurbiprofen axetil fat emulsion injection, preparation method and application
CN112755196B (en) Donepezil Ji Lizi liquid
JP2023536371A (en) Ibuprofen ester derivative and its emulsion preparation
CA3200105A1 (en) Thc derivatives, oral dosage forms comprising same, uses thereof for treating diseases and disorders and synthesis thereof
EP4243773A1 (en) Solid dispersions of cannabidiol
CN114644673A (en) Estradiol derivative, preparation method and medical application thereof
US20160362402A1 (en) Novel compounds having anti-allodynic and antihyperalgesic activity