CN114728920B

CN114728920B - Vortioxetine prodrug and application thereof

Info

Publication number: CN114728920B
Application number: CN202080080988.0A
Authority: CN
Inventors: 张玲; 蒋钰; 陈帅; 张长青; 周珺; 万泽红
Original assignee: Suzhou Enhua Biomedical Technology Co ltd
Current assignee: Suzhou Enhua Biomedical Technology Co ltd
Priority date: 2019-12-10
Filing date: 2020-12-10
Publication date: 2023-07-21
Anticipated expiration: 2040-12-10
Also published as: CN114728920A; WO2021115372A1

Abstract

A vortioxetine prodrug and application thereof have a compound structure shown in a formula (A), and the compound can be used for preparing and treatingA medicament for treating neuropsychiatric diseases,

Description

Vortioxetine prodrug and application thereof

the present application claims priority from chinese patent application 201911254627.9, chinese patent application 2019/12/10. The present application refers to the entirety of the above-mentioned chinese patent application.

Technical Field

The invention belongs to the field of medicines, and particularly relates to a novel vortioxetine prodrug, a preparation method thereof, a composition containing the compound and application thereof.

Background

Major depressive disorder (Major depressive disorder, MDD) is a series of depressive symptoms, mainly sustained spontaneous depressed mood, which plague patients' ability to work, sleep, learn, eat and enjoy the current pleasure. Patients with major depression may experience pessimistic boredom, disfigurement, hallucinations, hypofunction, and serious suicidal attempts, and even suicidal behavior. A serious threat to human health is therefore a great deal of attention must be paid.

Vortioxetine (vortixoxetine) is a new drug for the treatment of depression, developed by the combination of danish northbound (Lundbeck) and martial arts (Takeda) and approved by the FDA for use in the treatment of MDD at 9 and 30 days 2013 under the trade name brinellix. Common adverse reactions of vortioxetine are gastrointestinal side effects such as nausea, constipation and vomiting, which are the major side effects of SSRIs antidepressants. The incidence of adverse reactions in the 5mg vortioxetine group was high for nausea (19.4 vs 9.4%) and diarrhea (11.4 vs 7.0%) compared to placebo, with an increase in the incidence of gastrointestinal side effects with further increases in the doses administered ([ J ]. Pharmacology & therapeutics,2015, 145:43-57).

WO2017215636A1 discloses a prodrug of vortioxetine with the compound structure shown below:

due to the existence of a Blood Brain Barrier (BBB), the higher the fat-soluble solute, the faster the solute can enter brain tissue through the blood brain barrier, for example, according to the law, certain CNS drugs can be modified, and the fat-soluble side chain is introduced to be made into fat-soluble pro-drugs, so that the membrane penetration capacity is improved, the drugs can enter brain tissue more easily through the BBB, and the brain/blood ratio of the drugs is increased, so that the drugs have central nervous system targeting drug delivery effect (J Drug discovery today,2007, 12 (1-2): 54-61); however, the relationship of the three of improving the penetration ability of the prodrug to the blood brain barrier, reducing the breaking rate of the modified chain of the prodrug in the plasma, and rapidly metabolizing the prodrug to the original drug at the target position is a difficulty in modifying the prodrug. As described in page 1 of patent WO2017215636A1 specification 21, the rate of metabolizing vortioxetine prodrug to proto-drug by drug metabolizing enzyme is changed along with the change of lipid solubility of modified side chain, so that a great deal of work is required to find out modified chain which breaks down slowly in plasma and breaks down rapidly in targeted drug release position (such as brain tissue), thus the modified vortioxetine prodrug can make proto-drug vortioxetine release into brain rapidly, reduce drug concentration in plasma and achieve the effects of quick action and reduction of adverse reaction in gastrointestinal tract.

In summary, the modification of vortioxetine prodrugs remains an important focus in the field of depression treatment, and vortioxetine prodrugs that can significantly reduce the dosage and increase the brain penetration rate are the hot spot of current research.

Disclosure of Invention

On one hand, the compound provided by the invention is used for connecting vortioxetine with different fat-soluble side chains so as to improve the cerebral blood ratio of vortioxetine, reduce the dosage of administration, enhance the drug effect and reduce the side effect; on the other hand, valproic acid, gabapentin and pregabalin are respectively connected with vortioxetine in a chemical mode for drug combination, so that the lipid solubility of the prodrug can be increased, the brain penetration of vortioxetine is improved, and two drugs with different mechanisms can play a role in drug synergism in mammals and/or human bodies.

In one aspect, the present invention provides a compound of formula (A), a stereoisomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof,

wherein:

x-is a single bond "-" or- (CH) ₂ )n ₁ -any one of the following;

R ₁ and R is ₂ Each independently selected from C optionally substituted with halogen, amino, hydroxy or cyano _1-3 Any of alkyl, or hydrogen;

R ₃ selected from C optionally substituted by halogen, hydroxy, cyano or amino _6-10 Is a group consisting of an aryl group,any one of them;

R ₄ and R is ₅ Each independently selected from hydrogen, amino, C optionally substituted with halogen, amino or hydroxy _6-15 Or C optionally substituted by amino, hydroxy, halogen or cyano _1-5 Any one of the alkyl groups of (a);

n ₁ is any integer between 1 and 3;

a and b are each independently any integer between 0 and 2.

In another aspect, the present invention provides a method for preparing a compound of formula (a), a stereoisomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, according to the invention, comprising:

the compound of the general formula (III) and the compound of the general formula (IV) are subjected to substitution reaction to obtain a compound of the general formula (A);

wherein:

-X-，R ₁ ，R ₂ and R is ₃ Each independently as defined above.

In yet another aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (a) of the present invention, a stereoisomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

In a further aspect, the present invention provides the use of a compound of formula (a), a stereoisomer thereof, a tautomer thereof or a pharmaceutically acceptable salt or a pharmaceutical composition thereof according to the invention in the manufacture of a medicament for the treatment of a neuropsychiatric disease.

In particular, the neuropsychiatric disease is selected from one or more of depression, anxiety, sleep disorders, neurodegenerative diseases, cognitive disorders, bipolar disorders, post-traumatic stress syndrome, addictive disorders, withdrawal syndrome or attention deficit disorder, preferably one or more of depression, anxiety, schizophrenia, sleep disorders, cognitive disorders, neurodegenerative diseases or bipolar disorders.

Detailed Description

General terms and definitions

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. If there is a conflict, the present application provides definitions. When trade names are presented herein, it is intended to refer to their corresponding commercial products or active ingredients thereof. All patents, published patent applications, and publications cited herein are incorporated by reference.

The terms "comprising," "including," "having," "containing," or "involving," and other variations thereof herein, are inclusive or open-ended and do not exclude additional unrecited elements or method steps. Those skilled in the art will appreciate that the terms such as "comprising" encompass "consisting of.

In addition, unless explicitly stated otherwise, the description as used in this disclosure of the present invention is to be understood in a broad sense as "each independently" or "each independently" and refers to the fact that, in different groups, specific items expressed between the same symbols do not affect each other, or that, in the same groups, specific items expressed between the same symbols do not affect each other.

The term "one or more" or similar expression "at least one" may denote, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more.

When lower and upper limits of a range of values are disclosed, any number and any range encompassed within the range are specifically disclosed. In particular, each numerical range of values disclosed herein is to be understood as meaning every number and range that is encompassed within the broader range.

The expression m-n as used herein refers to the range of m to n and the sub-ranges consisting of the individual point values therein as well as the individual point values. For example, the expression "C ₆ -C ₁₅ "OR" C _6-15 "ranges from 6 to 15 carbon atoms are to be understood as also covering any subrange therein as well as every point value, e.g. C ₆ -C ₁₅ 、C ₇ -C ₁₄ 、C ₈ -C ₁₃ 、C ₉ -C ₁₂ 、C ₆ -C ₁₄ 、C ₆ -C ₁₃ 、C ₇ -C ₁₅ 、C _10-15 Etc. and C ₆ 、C ₇ 、C ₈ 、C ₉ 、C ₁₀ 、C ₁₁ 、C ₁₂ Etc. For example, the expression "C ₁ -C ₃ "OR" C _1-3 "also should be understood in a similar manner, for example, any subrange and point value contained therein, e.g., C ₁ -C ₃ 、C ₁ -C ₂ 、C ₂ -C ₃ Etc. and C ₁ 、C ₂ 、C ₃ Etc. Also for example, the expression "C ₁ -C ₅ "OR" C ₁ - ₅ "ranges from 1 to 5 carbon atoms are to be understood as also covering any subrange therein as well as every point value, e.g. C ₂ -C ₅ 、C ₃ -C ₄ 、C ₁ -C ₂ 、C ₁ -C ₃ 、C ₁ -C ₄ 、C ₁ -C ₅ Etc. and C ₁ 、C ₂ 、C ₃ 、C ₄ 、C ₅ Etc.

The term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

The terms "substituted" and "substituted" refer to the replacement of one or more (e.g., one, two, three, or four) hydrogens on the designated atom with a selection from the indicated group, provided that the designated atom's normal valency is not exceeded, and that the substitution forms a stable compound. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds. When it is described that a substituent is absent, it is understood that the substituent may be one or more hydrogen atoms, provided that the structure is such that the compound is stable. When it is described that each carbon atom in a group can optionally be replaced by a heteroatom, provided that the normal valency of all atoms in the group in the current case is not exceeded, and stable compounds are formed.

A substituent may be unsubstituted (unsubstituted) or may be substituted if the substituent is described as "optionally substituted. For example, R ₁ Defined as "C optionally substituted with halogen, amino, hydroxy or cyano _1-3 Alkyl "of (C) represents R ₁ C being substituted by halogen, amino, hydroxy or cyano _1-3 Or unsubstituted C _1-3 When R is an alkyl group of ₁ Expressed as C only _1-3 C when alkyl is an alkyl group _1-3 Is unsubstituted (unsubstituted), R ₂ 、R ₃ 、R ₄ And R is ₅ Etc. are defined as R ₁ . If an atom or group is described as being optionally substituted with one or more of the list of substituents, then one or more hydrogens on that atom or group may be replaced with an independently selected, optional substituent. When the substituent is oxo (i.e., =o), it means that two hydrogen atoms are replaced.

As used herein, unless indicated, the point of attachment of a substituent may be from any suitable position of the substituent. When the bond of a substituent is shown as a bond through the ring connecting two atoms, then such substituent may be bonded to any ring-forming atom in the substitutable ring.

When any variable (e.g., R), and the variable with a label (e.g., R ₁ 、R ₂ 、R ₃ 、R ₄ 、R ₅ 、n ₁ A, b, etc.) are independent in each case at each occurrence of their definition when they occur more than once in the composition or structure of the compound. For example, if a group is substituted with 0, 1,2, 3 or 4R substituents, the group may optionally be substituted with up to four R substituents, and the options for each R substituent in each case are independent of each other.

The term "halogen" or "halo" is understood to mean a fluorine (F), chlorine (C1), bromine (Br) or iodine (I) atom, preferably a fluorine, chlorine, bromine atom.

The term "alkyl" refers to a straight or branched saturated aliphatic hydrocarbon group consisting of carbon and hydrogen atoms, which is attached to the remainder of the molecule by a single bond. "alkyl" may have 1 to 15 carbon atoms, i.e. "C ₁ -C ₁₅ Alkyl ", e.g. C _1-4 Alkyl, C _1-3 Alkyl, C _1-2 Alkyl, C ₃ Alkyl, C ₄ Alkyl, C _1-5 Alkyl, C _6-15 An alkyl group. Non-limiting examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-ethylpropyl, 1, 2-dimethylpropyl, neopentyl, 1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 2, 3-dimethylbutyl, 1, 3-dimethylbutyl, or 1, 2-dimethylbutyl, or isomers thereof.

The term "aryl" means a monocyclic, bicyclic, and tricyclic carbocyclic ring system containing 6 to 14 ring atoms, or 6 to 12 ring atoms, or 6 to 10 ring atoms, wherein at least one ring is aromatic. The aryl group is typically, but not necessarily, attached to the parent molecule through an aromatic ring of the aryl group. Examples of aryl groups may include phenyl, naphthyl, and anthracene. The aryl group is optionally substituted by one or more of the substituted groups described herein.

The term "pharmaceutically acceptable" refers to substances which, within the scope of normal medical judgment, are suitable for use in contact with the tissues of a patient without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit-to-benefit ratio, and effective for their intended use.

The compounds of the invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-) -and (+) -enantiomers, (R) -and (S) -enantiomers, diastereomers, (D) -isomers, (L) -isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of the present invention. In certain embodiments, preferred compounds are those isomeric compounds that exhibit more biological activity. Purified or partially purified isomers and stereoisomers, or racemic or diastereomeric mixtures of the compounds of the invention are also included within the scope of the invention. Purification and isolation of such materials can be accomplished by standard techniques known in the art.

The term "pharmaceutically acceptable carrier" refers to those substances which have no significant irritating effect on the organism and which do not impair the biological activity and properties of the active compound. "pharmaceutically acceptable carrier" includes, but is not limited to, glidants, sweeteners, diluents, preservatives, dyes/colorants, flavoring agents, surfactants, wetting agents, dispersing agents, disintegrants, stabilizers, solvents or emulsifiers.

The terms "administration" or "administering" and the like refer to a method that may enable delivery of a compound or composition to a desired biological site of action. These methods include, but are not limited to, oral or parenteral (including intraventricular, intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular injection or infusion), topical, rectal administration, and the like. In particular for injection or oral administration.

As used herein, the term "treating" includes alleviating, alleviating or ameliorating a disease or symptom, preventing other symptoms, ameliorating or preventing underlying metabolic factors of a symptom, inhibiting a disease or symptom, e.g., preventing the development of a disease or symptom, alleviating a disease or symptom, promoting alleviation of a disease or symptom, or halting signs of a disease or symptom, and extends to including prevention. "treating" also includes achieving therapeutic benefit and/or prophylactic benefit. Therapeutic benefit refers to eradication or amelioration of the condition being treated. In addition, therapeutic benefit is achieved by eradicating or ameliorating one or more physiological symptoms associated with the underlying disease, although the patient may still have the underlying disease, an amelioration of the patient's disease may be observed. Prophylactic benefit means that the patient is using the composition to prevent a risk of a disease, or is taking the patient when one or more physiological conditions of the disease are present, although the disease has not yet been diagnosed.

The term "comorbidity" means that two diseases co-exist, or one disease accompanies another, or (simultaneously produced/accompanied) the disease, e.g., two sets of symptoms are considered separately, all meeting respective diagnostic criteria.

The term "active ingredient", "therapeutic agent", "active substance" or "active agent" refers to a chemical entity that is effective in treating or preventing a disorder, disease or condition of interest.

The term "pharmaceutically acceptable salt" or "pharmaceutically acceptable salt" refers to salts with pharmaceutically acceptable non-toxic bases or acids, including inorganic or organic bases and inorganic or organic acids. Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof. Suitable acid addition salts are formed from acids that form pharmaceutically acceptable salts; suitable base addition salts are formed from bases which form pharmaceutically acceptable salts; methods for preparing pharmaceutically acceptable salts of the compounds of the invention are known to those skilled in the art.

The term "effective amount", "therapeutically effective amount" or "prophylactically effective amount" with respect to a drug, drug unit or active ingredient refers to a sufficient amount of a drug or agent that is acceptable for side effects but achieves the intended effect. Determination of an effective amount varies from person to person, depending on the age and general condition of the individual, and also on the particular active substance, a suitable effective amount in an individual can be determined by one skilled in the art from routine experimentation.

As used herein, "individual" includes human or non-human animals. Exemplary human individuals include human individuals (referred to as patients) or normal individuals suffering from a disease (e.g., a disease described herein). "non-human animals" in the context of the present invention include all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, domestic animals and/or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).

Herein, "X" and "-X-" are both denoted as the same particular group, which may be used interchangeably.

In the present invention, when the bond "- - -" described is absent, the substituent to which it is attached and the position to which it is substituted are directly attached, for example, when R ₃ Is thatWhen any one of the radicals "- - - - -" is absent, R ₄ And R is ₅ Each independently directly linked to a corresponding substituted position; for example, C _3-7 R when cycloalkyl is directly linked to the corresponding substituted position ₃ Can be->Etc.

The following detailed description is intended to illustrate non-limiting embodiments so that others skilled in the art may more fully understand the invention's solution, its principles and its practical application, to thereby modify and practice the invention in many forms best suited to the requirements of a particular use.

Compounds of formula (A)

wherein:

x-is a single bond "-" or- (CH) ₂ )n ₁ -any one of the following;

n ₁ is any integer between 1 and 3;

a and b are each independently any integer between 0 and 2.

In a preferred embodiment of the invention, wherein:

x-is a single bond "-" or- (CH) ₂ )n ₁ -any one of the following;

R ₁ and R is ₂ Each independently selected from C _1-3 Any one of alkyl or hydrogen;

R ₃ selected from C _6-10 Is a group consisting of an aryl group,any one of them;

R ₄ and R is ₅ Each independently selected from hydrogen, amino, C _6-15 Or C optionally substituted by amino _1-5 Any one of the alkyl groups of (a);

n ₁ Is any integer between 1 and 3;

a and b are each independently any integer between 0 and 2.

In one embodiment of the invention, the n ₁ Is any integer of 1,2 or 3; each of a and b is independently any integer of 0,1 or 2.

In one embodiment of the invention, the halogen is selected from fluorine, chlorine, bromine and iodine. In a more preferred embodiment, the halogen is selected from fluorine, chlorine and bromine. Such as fluorine or chlorine.

In a preferred embodiment of the present invention, said n ₁ 1 or 2, preferably 1; a. b are each independently 0,1 or 2, preferably 1.

In another embodiment of the present invention, the compound represented by the general formula (A), a stereoisomer thereof, a tautomer thereof or a pharmaceutically acceptable salt thereof is further selected from any one of the compounds represented by the general formula (A-1), (A-2) or (A-3) shown below:

wherein:

x-is a single bond "-" or- (CH) ₂ )n ₁ -any one of the following;

R ₃ selected from C optionally substituted by halogen, hydroxy, cyano or amino _6-10 Aryl of (a);

n ₁ is any integer between 1 and 3;

a and b are each independently any integer between 0 and 2.

In a preferred embodiment of the invention, -X-is a single bond "-" or- (CH) ₂ )n ₁ -any one of the following; r is R ₁ And R is ₂ Each independently selected from C _1-3 Any one of alkyl or hydrogen; r is R ₃ Selected from C _6-10 Aryl of (a); r is R ₄ And R is ₅ Each independently of the otherIs selected from hydrogen, amino, C _6-15 Or C optionally substituted by amino _1-5 Any one of the alkyl groups of (a); n is n ₁ Is any integer between 1 and 3; a and b are each independently any integer between 0 and 2.

In a preferred embodiment of the invention, the n ₁ 1 or 2, preferably 1; a. b are each independently 0, 1, 2, preferably 1.

In a preferred embodiment of the invention, the C is optionally substituted by halogen, amino, hydroxy or cyano _1-3 The alkyl group of (2) is selected from C _1-3 Alkyl group of said C _1-3 The alkyl group of (a) is selected from any one of methyl, ethyl or propyl, preferably any one of methyl or ethyl.

In a preferred embodiment of the invention, the C is optionally substituted with amino, hydroxy, halogen or cyano _1-5 Is selected from C optionally substituted by amino _1-5 An alkyl group of (C) optionally substituted with an amino group _1-5 Is selected from any one of methyl, ethyl, propyl, butyl or amyl optionally substituted by amino; preferably any one of methyl, ethyl, propyl or butyl optionally substituted with amino; more preferably any one of methyl, ethyl or propyl optionally substituted with amino.

In a more preferred embodiment of the present invention, the propyl group is selected from n-propyl (n-Pr, -CH) ₂ CH ₂ CH ₃ ) Or isopropyl ((i-Pr, -CH (CH) ₃ ) ₂ ). The butyl is selected from n-butyl (n-Bu, -CH) ₂ CH ₂ CH ₂ CH ₃ ) Isobutyl (i-Bu, -CH) ₂ CH(CH ₃ ) ₂ ) Sec-butyl (s-Bu, -CH (CH) ₃ )CH ₂ CH ₃ ) Or tert-butyl (t-Bu, -C (CH) ₃ ) ₃ ). The amyl group is selected from n-amyl (-CH) ₂ CH ₂ CH ₂ CH ₂ CH ₃ ) 2-pentyl (-CH (CH) ₃ )CH ₂ CH ₂ CH ₃ ) 3-pentyl (-CH (CH) ₂ CH ₃ ) ₂ ) 2-methyl-2-butyl (-C (CH) ₃ ) ₂ CH ₂ CH ₃ ) 3-methyl-2-butyl (-CH (CH) ₃ )CH(CH ₃ ) ₂ ) 3-methyl-1-butyl (-CH) ₂ CH ₂ CH(CH ₃ ) ₂ ) Or 2-methyl-1-butyl (-CH) ₂ CH(CH ₃ )CH ₂ CH ₃ )。

In a preferred embodiment of the invention, the C is optionally substituted by halogen, amino or hydroxy _6-15 The alkyl group of (2) is selected from C _6-15 Alkyl group of said C _6-15 The alkyl group of (2) is selected from C _6-15 Straight-chain alkyl of (C) is preferred _9-15 Straight chain alkyl of (a).

In a preferred embodiment of the invention, the C is optionally substituted with halogen, hydroxy, cyano or amino _6-10 Aryl of (2) is selected from C _6-10 Aryl of (C), said C _6-10 Is selected from any one of phenyl or naphthyl; more preferably phenyl.

In a preferred embodiment of the invention, said R ₁ And R is ₂ Each independently selected from C optionally substituted with halogen, amino, hydroxy or cyano _1-3 Any of alkyl, or hydrogen; preferably C _1-3 Any one of alkyl or hydrogen; more preferably any one of hydrogen, methyl or ethyl; in particular, hydrogen or methyl is preferred.

In a preferred embodiment of the invention, said R ₃ Selected from C optionally substituted by halogen, hydroxy, cyano or amino _6-10 Is a group consisting of an aryl group,any one of them; wherein the C is optionally substituted with halogen, hydroxy, cyano or amino _6-10 Aryl of (2) is preferably C _6-10 More preferably a phenyl group or a naphthyl group, and particularly preferably a phenyl group.

In a preferred embodiment of the invention, R ₄ And R is ₅ Each independently selected from hydrogen, amino, C optionally substituted with halogen, amino or hydroxy _6-15 Or C optionally substituted by amino, hydroxy, halogen or cyano _1-5 Any one of the alkyl groups of (a); more preferably hydrogen, amino, C _6-15 Or C optionally substituted by amino _1-5 Any one of the alkyl groups of (a); in particular, R ₄ And R is ₅ Each independently is preferably hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, -CH ₂ NH ₂ ，-CH ₂ CH ₂ NH ₂ ，-CH ₂ CH ₂ CH ₂ NH ₂ ，-(CH ₂ ) ₈ CH ₃ ，-(CH ₂ ) ₉ CH ₃ ，-(CH ₂ ) ₁₂ CH ₃ Or- (CH) ₂ ) ₁₃ CH ₃ Any one of the following.

In a more preferred embodiment of the invention, R ₄ And R is ₅ Each independently is preferably hydrogen, methyl, n-propyl, isopropyl, -CH ₂ CH ₂ NH ₂ ，-(CH ₂ ) ₈ CH ₃ ，-(CH ₂ ) ₉ CH ₃ ，-(CH ₂ ) ₁₂ CH ₃ Or- (CH) ₂ ) ₁₃ CH ₃ Any one of the following.

In a preferred embodiment of the invention, the n ₁ Is any integer between 1 and 3; preferably any one integer of 1,2 or 3, more preferably 1.

In a preferred embodiment of the present invention, each of a and b is independently any integer between 0 and 2, preferably any integer of 0,1 or 2, more preferably 1.

In the present invention, it is expressed as "C" only _1-3 Alkyl group "," C _6-15 Alkyl group "," C _6-15 Straight chain alkyl groups of (C) _6-10 Aryl "or the like is unsubstituted or unsubstituted.

In a preferred further embodiment of the present invention, there is provided a compound represented by the formula (A), a stereoisomer thereof, a tautomer thereof or a pharmaceutically acceptable salt thereof,

Wherein:

x-is a single bond "-" or- (CH) ₂ )n ₁ -any one of n ₁ 1 is shown in the specification;

R ₁ and R is ₂ Each independently selected from any one of hydrogen, methyl or ethyl;

R ₃ is selected from the group consisting of phenyl groups,any one of them;

R ₄ and R is ₅ Each independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, -CH ₂ NH ₂ ，-CH ₂ CH ₂ NH ₂ ，-CH ₂ CH ₂ CH ₂ NH ₂ ，-(CH ₂ ) ₈ CH ₃ ，-(CH ₂ ) ₉ CH ₃ ，-(CH ₂ ) ₁₂ CH ₃ Or- (CH) ₂ ) ₁₃ CH ₃ Any one of them;

a is 1, and b is 1.

In a preferred embodiment of the present invention, the-X-is a single bond "-" or- (CH) ₂ )n ₁ -any one of n ₁ 1 is shown in the specification; r is R ₁ And R is ₂ Each independently selected from any one of hydrogen or methyl; r is R ₃ Is selected from the group consisting of phenyl groups,any one of them; r is R ₄ And R is ₅ Each independently selected from hydrogen, methyl, n-propyl, isopropyl, -CH ₂ CH ₂ NH ₂ ，-(CH ₂ ) ₈ CH ₃ ，-(CH ₂ ) ₉ CH ₃ ，-(CH ₂ ) ₁₂ CH ₃ Or- (CH) ₂ ) ₁₃ CH ₃ Any one of them; a is 1, and b is 1.

In a preferred further embodiment of the present invention, the exemplary compounds of the general formula (a), stereoisomers thereof, tautomers thereof or pharmaceutically acceptable salts thereof are as follows:

table 1 exemplary Compounds of the Compounds of formula (A)

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In a preferred further embodiment of the present invention, the compound represented by the general formula (a), a stereoisomer thereof, a tautomer thereof or a pharmaceutically acceptable salt thereof is preferably a compound represented by the following formula (a):

Among the pharmaceutically acceptable salts of the compounds of the present invention, the pharmaceutically acceptable salts may be selected from the group consisting of hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, tartrate, maleate, fumarate, methanesulfonate, trifluoroacetate, gluconate, saccharate, benzoate, ethanesulfonate, benzenesulfonate or p-toluenesulfonate. Hydrobromide or trifluoroacetate is preferred.

In a preferred embodiment of the invention, the pharmaceutically acceptable salt is preferably trifluoroacetate salt.

In a preferred embodiment of the present invention, the pharmaceutically acceptable salts of compounds EP-0015-014 and E-P0015-015 are preferably trifluoroacetate salts, more preferably monotrifluoroacetate salts.

In an embodiment of the invention, the pharmaceutically acceptable salt is prepared from a compound, its stereoisomer or its tautomer, and its corresponding acid or base.

Process for preparing compounds of formula (A)

Preparation of Compounds of the general formula (A)

The present invention further provides a process for preparing a compound of formula (a), a stereoisomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, comprising:

Substitution reaction of the compound of formula (III) with a commercially available compound of formula (IV) gives a compound of formula (A).

In a preferred embodiment of the present invention, the compound of formula (A) is prepared by reacting a compound of formula (III) with a compound of formula (IV) in the presence of an inorganic base and an organic solvent. If under the protection of nitrogen, adding the compound of the general formula (III) and inorganic base such as cesium carbonate or potassium carbonate into a proper solvent such as DMF, and reacting for 0.5-3 hours at 40 ℃ to obtain the compound of the general formula (A).

In a preferred further embodiment of the invention, the compounds of the formula (IV) are prepared in the presence of a compound containing an amino group (e.g. -NH- ₂ ) In the case of (a), the compound of the general formula (a) can be prepared by reacting with an amino protecting reagent to protect an amino group contained in the compound of the general formula (IV), then reacting with the compound of the general formula (III), and then removing the protecting group. The amino protecting agent includes, but is not limited to, di-tert-butyl dicarbonate (Boc ₂ O)。

Preparation of Compounds of the general formula (III)

The invention further provides a process for the preparation of a compound of formula (III), comprising:

the compounds of formula (I) are reacted with commercially available compounds of formula (II) to produce compounds of formula (III).

In a preferred embodiment of the present invention, the compound of formula (I) is dissolved in a suitable organic solvent and triethylamine is reacted with the compound of formula (I) to prepare the compound of formula (III). For example, in Dichloromethane (DCM), the compound of formula (I), triethylamine and the compound of formula (II) are added and reacted at room temperature for 1-3 hours to give the compound of formula (III).

In the above-described embodiment of the production method of the present invention,

wherein:

x-is a single bond "-" or- (CH) ₂ )n ₁ -any one of the following;

n1 is any integer between 1 and 3;

a and b are each independently any integer between 0 and 2.

Pharmaceutical composition and pharmaceutical formulation

The present invention further provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (a), a stereoisomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

In a preferred embodiment of the invention, the pharmaceutical composition may be formulated in a conventional manner using one or more pharmaceutically acceptable carriers. Thus, the active compounds of the present invention may be formulated in dosage forms for oral, buccal, intranasal, parenteral (e.g. intravenous, intramuscular or subcutaneous) or rectal administration, or in dosage forms suitable for administration by inhalation or insufflation. The compounds of the invention may also be formulated in sustained release dosage forms.

In a preferred embodiment of the invention, an effective dose of a compound of the invention may be administered orally, e.g., with an inert diluent or with some carrier. According to some embodiments of the invention, the compounds of the invention may be encapsulated in gelatin capsules or compressed into tablets. For the purpose of oral treatment, the compounds of the present invention may be used together with excipients and in the form of tablets, troches, capsules, suspensions, syrups and the like. According to embodiments of the present invention, the above formulations should contain at least 0.5% (w/w) of the active compound of the present invention, but may vary depending on the particular dosage form, with 4% to about 70% by weight being convenient. The amount of active compound in such pharmaceutical compositions should be such that an appropriate dosage is achieved.

In a preferred embodiment of the invention, for oral administration, the active compounds according to the invention can be formulated, for example, by conventional means with pharmaceutically acceptable excipients, such as binders, fillers, lubricants, disintegrants or wetting agents, into tablets or capsules. The tablets may be coated by methods well known in the art. Liquid formulations for oral administration, such as may be employed in solution, syrup or suspension, or volatilized as a dry product, are reconstituted with water or other suitable carrier prior to use. Such liquid formulations may be prepared by conventional means using pharmaceutically acceptable additives such as suspending agents, emulsifying agents, non-aqueous vehicles and preservatives.

In a preferred embodiment of the invention, when the active compounds of the invention or stereoisomers, tautomers or pharmaceutically acceptable salts thereof, are for parenteral administration, the compounds provided herein may be combined with sterile water or an organic medium to form an injectable solution or suspension.

In a preferred embodiment of the invention, the active compounds of the invention or stereoisomers, tautomers or pharmaceutically acceptable salts thereof may be formulated in rectal compositions, such as suppositories or retention enemas, e.g. containing conventional suppository bases such as cocoa butter or other glycerides.

Therapeutic uses and methods

In a further aspect, the invention provides the use of a compound of the invention, a stereoisomer thereof, a tautomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the invention, in the manufacture of a medicament for the treatment of a neuropsychiatric disease.

In another aspect, the invention also provides a method of treating a neuropsychiatric disease comprising administering to a subject (e.g., a mammalian subject, such as a human) in need thereof a compound of the invention, a stereoisomer thereof, a tautomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the invention.

In one embodiment, the neuropsychiatric disease is selected from any one or more of schizophrenia, depression, anxiety, sleep disorders, neurodegenerative diseases, cognitive disorders, bipolar disorders, post-traumatic stress syndrome, addictive disorders, withdrawal syndrome or attention deficit disorder, preferably any one or more of depression, cognitive disorders, anxiety, schizophrenia, sleep disorders, neurodegenerative diseases or bipolar disorders; more preferably depression or cognitive disorders. In a preferred embodiment, the neuropsychiatric disease is depression.

The invention also provides the use of a compound of formula (a), a stereoisomer thereof, a tautomer thereof or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of depression, wherein the medicament optionally comprises one or more additional agents which modulate the nervous system of a mammal or alleviate depression.

In a preferred embodiment of the present invention, the present invention also provides the use of a compound of formula (a), a stereoisomer thereof, a tautomer thereof or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of depression.

In a preferred embodiment of the invention, the depression includes, but is not limited to, light to major depression, preferably medium to major depression, in compliance with relevant diagnostic criteria of WHO ICD-10 (International Classification of diseases), american DSM-5 (Manual of diagnosis and statistics of mental disorders) and CCMD-3 (national Classification of mental diseases and diagnostic criteria).

In a preferred embodiment of the invention, the depression is selected from the group consisting of major depression, moderate depression, major depression, depression with other mental disorders, and recurrent depression; in a more preferred embodiment, the depression is selected from the group consisting of moderate depression, major depression, depression accompanied by other mental disorders, and recurrent depression.

In one embodiment of the invention, the depression may co-treat at least one psychotic disorder.

In a preferred embodiment of the invention, the mental disorder is selected from one or more of a sleep disorder, a cognitive disorder, an anxiety disorder, a obsessive-compulsive disorder, an affective disorder or a personality disorder, preferably a cognitive disorder, an anxiety disorder or a sleep disorder, more preferably a cognitive disorder.

In a preferred embodiment of the invention, the depression comprises co-morbid other psychotic disorders, including but not limited to depression co-morbid cognitive disorders, depression co-morbid sleep disorders, depression co-morbid anxiety disorders, depression co-morbid sensory disorders, depression co-morbid social disorders or depression co-patient lattice disorders, and the like, preferably depression co-morbid cognitive disorders, depression co-morbid anxiety disorders or depression co-morbid sleep disorders; more preferably a depressive co-morbid cognitive disorder.

In embodiments of the invention, the co-morbid other mental disorders also refer to the concomitant other mental disorders.

In a preferred further embodiment of the present invention, the use of a compound of formula (a), a stereoisomer thereof, a tautomer thereof or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising a compound of formula (a), a stereoisomer thereof, a tautomer thereof or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of depression or depression with other psychotic disorders, preferably depression with cognitive disorders.

Description of the embodiments

The invention provides a compound shown in a general formula (A), a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof,

it is characterized in that the method comprises the steps of,

x-is a single bond "-" or- (CH) ₂ )n ₁ -any one of the following;

R ₃ is that

R ₄ And R is ₅ Each independently selected from hydrogen, amino, unsubstituted C _6-15 C substituted or unsubstituted by amino _1-5 Alkyl of C _3-7 Cycloalkyl, C _6-10 Any one of aryl or absent;

bond "- -" is any one of a single bond "-" or absent;

n ₁ is any integer between 1 and 3.

In a further preferred embodiment of the invention, n1 is 1 or 2, preferably n1 is 1.

In a preferred embodiment of the present invention, the compound of formula (a), a stereoisomer thereof, a tautomer thereof or a pharmaceutically acceptable salt thereof, wherein:

x-is a single bond "-" or- (CH) ₂ )n ₁ -any one of the following;

R ₁ and R is ₂ Each independently selectFrom C _1-3 Any one of alkyl or hydrogen;

R ₄ and R is ₅ Each independently selected from hydrogen, amino, unsubstituted C _6-15 C substituted or unsubstituted by amino _1-5 Any one of the alkyl groups of (a);

n ₁ is any integer between 1 and 3;

a and b are each independently any integer between 0 and 2.

In a further preferred embodiment of the invention, said n1 is 1 or 2, preferably n1 is 1; a and b are each independently 0, 1 or 2, preferably 1.

In a preferred further embodiment of the present invention, there is provided a compound represented by the formula (A-1) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof,

wherein:

x-is a single bond "-" or- (CH) ₂ )n ₁ -any one of the following;

bond "- -" is any one of a single bond "-" or absent;

n ₁ is any integer between 1 and 3.

In a further preferred embodiment of the invention, said n ₁ Is 1 or 2, preferably n ₁ 1.

In a further preferred embodiment of the invention, the C _1-3 The alkyl group of (a) is selected from methyl, ethyl or propyl, preferably methyl, ethyl, more preferably methyl.

In a further preferred embodiment of the invention, the amino-substituted or unsubstituted C _1-5 Is methyl, ethyl, propyl, butyl or pentyl substituted or unsubstituted with amino.

In a further preferred embodiment of the invention, the amino-substituted C _1-5 Methyl, ethyl or propyl substituted with amino groups including, but not limited to, e.g., -CH ₂ -NH ₂ ，-(CH ₂ ) ₂ -NH ₂ ，-(CH ₂ ) ₃ -NH ₂ 。

In a further preferred embodiment of the present invention, the unsubstituted C _1-5 Preferably methyl, ethyl, propyl or butyl.

In a further preferred embodiment of the present invention, the unsubstituted C _6-15 Preferably C _6-15 More preferably C _9-15 Straight chain alkyl of (a).

In a further preferred embodiment of the invention, the C _3-7 Is selected from unsubstituted five-membered cycloalkyl or six-membered cycloalkyl; the C is _6-10 Preferably unsubstituted phenyl or naphthyl, more preferably unsubstituted phenyl.

In a preferred embodiment of the present invention, in the compound represented by the general formula (A) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, -X-is a single bond "-" or- (CH) ₂ )n ₁ -any one of the following; the C is _1-3 Is selected from any one of methyl or ethyl; the amino-substituted or unsubstituted C _1-5 Is selected from any one of methyl, ethyl, propyl or butyl substituted or unsubstituted by amino; unsubstituted C _6-15 Alkyl of (2)Selected from C _6-15 A linear alkyl group of (a); the C is _3-7 Is selected from any one of unsubstituted five-membered cycloalkyl or six-membered cycloalkyl; the C is _6-10 Is selected from any one of unsubstituted phenyl or naphthyl; and when R is ₄ And R is ₅ Each independently is C _3-7 Cycloalkyl or C of (C) _6-10 In the case of any one of the aryl groups of (a), the bond "- - -" is absent.

In another preferred embodiment of the present invention, in the compound represented by the general formula (A) or a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof, -X-is a single bond "-" or- (CH) ₂ )n ₁ -any one of the following; r is R ₁ And R is ₂ Each independently selected from any one of hydrogen, methyl or ethyl; r is R ₄ And R is ₅ Each independently selected from hydrogen, methyl, ethyl, propyl, butyl, -CH ₂ NH ₂ ，-CH ₂ CH ₂ NH ₂ Cyclohexane, phenyl, - (CH) ₂ ) ₉ CH ₃ ，-(CH ₂ ) ₁₃ CH ₃ Or absent; and when R is ₃ And R is ₄ Each independently is C _3-7 Cycloalkyl or C of (C) _6-10 In the absence of any one of the aryl groups of (a), bond "- - -" is absent; n is n ₁ 1 or 2.

In a preferred embodiment of the invention, the C _1-3 Is selected from any one of methyl or ethyl; the amino-substituted or unsubstituted C _1-5 Is selected from any one of methyl, ethyl, propyl or butyl substituted or unsubstituted by amino; unsubstituted C _6-15 The alkyl group of (2) is selected from C _6-15 A linear alkyl group of (a); the C is _3-7 Is selected from any one of unsubstituted five-membered cycloalkyl or six-membered cycloalkyl; the C is _6-10 Is selected from any one of unsubstituted phenyl or naphthyl; and when R is ₄ And R is ₅ Each independently is C _3-7 Cycloalkyl or C of (C) _6-10 In the case of any one of the aryl groups of (a), the bond "- - -" is absent.

In the present inventionIn a preferred embodiment, said R ₁ And R is ₂ Each independently selected from any one of hydrogen, methyl or ethyl; the R is ₄ And R is ₅ Each independently selected from hydrogen, methyl, ethyl, propyl, butyl, -CH ₂ NH ₂ ，-CH ₂ CH ₂ NH ₂ Cyclohexane, phenyl, - (CH) ₂ ) ₉ CH ₃ ，-(CH ₂ ) ₁₃ CH ₃ Or absent; and when R is ₃ And R is ₄ Each independently is C _3-7 Cycloalkyl or C of (C) _6-10 In the absence of any one of the aryl groups of (a), bond "- - -" is absent; n is n ₁ 1 or 2.

In a preferred embodiment of the present invention, the compound represented by the general formula (a) is selected from any one of the compounds represented by the following:

Preferably->

In a preferred embodiment of the invention, the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, tartrate, maleate, fumarate, methanesulfonate, trifluoroacetate, gluconate, saccharate, benzoate, ethanesulfonate, benzenesulfonate or p-toluenesulfonate, preferably trifluoroacetate.

The present invention provides a process for preparing a compound of formula (a), a stereoisomer thereof, a tautomer thereof or a pharmaceutically acceptable salt thereof, comprising:

wherein:

-X-，R ₁ ，R ₂ ，R ₃ and bonds "- -" are each independently as defined above.

The invention provides a preparation method of a compound shown as a general formula (III), which comprises the following steps:

reacting a compound of the general formula (I) with a compound of the general formula (II) to prepare a compound of the general formula (III);

wherein:

R ₁ and R is ₂ Each independently as defined above.

The invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound represented by general formula (A), a stereoisomer thereof, a tautomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

The invention provides a use of a compound shown in a general formula (A), a stereoisomer, a tautomer or a pharmaceutically acceptable salt or a pharmaceutical composition thereof in preparing a medicament for treating depression or other mental disorders accompanied by depression, preferably cognitive disorders accompanied by depression.

The beneficial technical effects of the invention

The present invention provides a novel prodrug of vortioxetine, generally speaking, the central nervous system drug is truly a free drug that enters the brain, whereas known vortioxetine binds more to peripheral tissues. In the test examples of the present invention, vortioxetine as the prototype drug was directly administered to cynomolgus monkeys, and the vortioxetine prodrug was administered without substantially detecting the drug concentration of free vortioxetine in the brain, and could be seenThe medicine concentration of the free vortioxetine in the brain of the cynomolgus monkey is obviously improved, and the potential advantage of improving the medicine effect is achieved. In addition, the vortioxetine prodrug compound has the advantages of obviously improving the cerebral blood proportion, reducing the peripheral exposure, having the potential effect of improving the gastrointestinal side effect of vortioxetine and having obvious brain targeting administration effect; at the same time, the prodrug compounds have good pharmacokinetic properties, including a more gradual release profile of the drug in vivo than vortioxetine, and an in-brain exposure (AUC _0-last ) The method has the advantages of reducing the exposure of vortioxetine in blood plasma, improving the relative bioavailability, improving the intake of vortioxetine and the like, and has the potential effect of improving the drug effect.

Drawings

FIG. 1 AUC ratio (cerebral blood ratio, B/P) of vortioxetine and vortioxetine prodrug compound in rats.

Figure 2 total AUC exposure of vortioxetine and vortioxetine prodrug compounds in rats was compared.

Figure 3 comparison of relative bioavailability of vortioxetine and vortioxetine prodrug compounds in rats.

Figure 4 drug-time profile of vortioxetine and vortioxetine prodrug in plasma in rats.

Figure 5 drug-time profile of vortioxetine and vortioxetine prodrug in brain tissue in rats.

Figure 6 total AUC exposure of vortioxetine and vortioxetine prodrugs in cynomolgus monkeys.

Detailed Description

Embodiments of the present invention are described in detail below. The following examples are illustrative only and are not to be construed as limiting the invention. Unless otherwise indicated, the proportions, percentages, etc., referred to herein are by weight.

Synthetic examples

Example 1, 1- (isobutyryloxy) ethyl 4- (2- ((2, 4-dimethylphenyl) thio) phenyl) piperazine-1-carboxylic acid (Compound EP-0015-003)

(1) In a 50mL single-necked flask, 0.5g vortioxetine (1.68 mmol) and 5mL methylene chloride were added, followed by 510mg triethylamine (5.04 mmol) and 360mg 1-chloroethyl chloroformate (2.52 mmol), and the reaction was stirred at room temperature for 2 hours. After the reaction, 10mL of water and 15mL of methylene dichloride are added to extract an organic phase, the organic phase is decompressed and concentrated to obtain a crude product, and the crude product is separated by column chromatography (petroleum ether: ethyl acetate=10:1, v/v) to obtain yellow oily matter EP 0015-003-A550 mg, the yield is 81.1%,

¹ H NMR(400MHz，CDCl ₃ )δ7.36(d，J＝7.6Hz，1H)，7.15(s，1H)，7.10-7.00(m，3H)，6.91-6.87(m，1H)，6.65-6.60(m，1H)，6.53(d，J＝8.0Hz，1H)，3.75-3.66(m，4H)，3.08-3.03(m，4H)，2.36(s，3H)，2.31(s，3H)，1.84(d，J＝8.0Hz，3H)；LCMS

[M+H] ⁺ 405.0。

(2) In a 50mL single-necked flask, 150mg of EP0015-003-A (0.37 mmol), 39mg of isobutyric acid (0.44 mmol), 362mg of cesium carbonate (1.11 mmol) and 3mL of N, N-dimethylformamide were added, and the mixture was heated and stirred at 40℃for 2 hours. After the reaction, 10mL of water and 15mL of ethyl acetate were added to extract an organic phase, and the organic phase was concentrated under reduced pressure to obtain a crude product, which was subjected to plate separation (petroleum ether: ethyl acetate=5:1, v/v) to obtain 90mg of a yellow oil, yield 53.2%, purity 99%.

¹ H NMR(400MHz，CDCl ₃ )δ7.36(d，J＝7.8Hz，1H)，7.19-7.13(m，1H)，7.11-6.99(m，3H)，6.91-6.79(m，2H)，6.52(m，1H)，3.67(m，4H)，3.03(m，4H)，2.63-2.49(m，1H)，2.36(s，3H)，2.31(s，3H)，1.51(d，J＝5.4Hz，3H)，1.18(m，6H)；LCMS

[M+H] ⁺ 457.1.0

Example 2, 1- (isobutyryloxyoxo) -2-methylpropyl 4- (2- ((2, 4-dimethylphenyl) thio) phenyl) piperazine-1-carboxylic acid (Compound EP-0015-004)

(1) In a 50mL single port flask, 200mg vortioxetine (0.671 mmol), triethylamine (0.805 mmol) and acetonitrile (2 mL) were added, cooled to 0deg.C, and 1-chloro-2-methylpropyl chloroformate (137 mg,0.805 mmol) dissolved in acetonitrile (2 mL) was added dropwise to the above solution. The reaction was stirred for 2h, monitored by tlc and the starting material was complete. The reaction solution was poured into 25mL of water and extracted twice with 20mL of EA (ethyl acetate). Na for organic layer ₂ SO ₄ Drying and concentrating to obtain crude product. Column chromatography with eluent (petroleum ether: ethyl acetate=5:1, v/v) gave 225mg ep0015-004-a as a white solid in 78% yield. LCMS [ M+H]+433.1

(2) 200mg (0.461 mmol) of the first-step product EP0015-004-A, 750mg (2.3 mmol) of cesium carbonate and 81mg (0.922 mmol) of isobutyric acid are dissolved in DMF (5 mL), the temperature is raised to 40 ℃ and the reaction is stirred for 3h, and the TLC monitors that the raw materials are basically reacted. After cooling to room temperature, 30mL of water was added and extracted twice with 25mL of ethyl acetate, and the organic layer was taken up in Na ₂ SO ₄ Drying and concentrating to obtain crude product. Column chromatography with eluent (petroleum ether: ethyl acetate=2:1, v/v) gave 80mg of white solid in 37% yield. HPLC purity: 99%.

¹ H NMR(400MHz，CDCl ₃ )：δ7.36(d，J＝8.0Hz，1H)，7.15(s，1H)，7.10-7.00(m，3H)，6.90-6.86(m，1H)，6.65(d，J＝8.0Hz，1H)，6.53-6.51(m，1H)，3.67(s，4H)，3.03(s，4H)，2.62-2.55(m，1H)，2.36(s，3H)，2.31(s，3H)，2.11-2.03(m，1H)，1.21-1.16(m，6H)，1.01-0.99(m，6H)；LCMS[M+H] ⁺ 485.1

Example 3, 1- (benzoyloxy) ethyl 4- (2- ((2, 4-dimethylphenyl) thio) phenyl) piperazine-1-carboxylic acid (Compound EP-0015-006)

150mg (0.371 mmol) of intermediate EP0015-003-A, 600mg (1.85 mmol) of cesium carbonate and 68mg (0.562 mmol) of benzoic acid are dissolved in DMF (3 mL), the temperature is raised to 45 ℃ and the reaction is stirred for 3h, and TLC monitors that the raw materials are basically reacted. Reaction solution addition 3After 0mL of water was extracted twice with 25mL of ethyl acetate, the organic layer was taken up with Na ₂ SO ₄ Drying and concentrating to obtain crude product. Column chromatography with eluent (petroleum ether: ethyl acetate=2:1, v/v) gave 40mg of white solid in 22% yield. HPLC purity: 99%.

¹ H NMR(400MHz，CDCl ₃ )：8.09-8.06(m，2H)，7.60-7.56(m，1H)，7.47-7.43(m，2H)，7.36(d，J＝8.0Hz，2H)，7.16-7.00(m，5H)，6.90-6.86(m，1H)，6.53-6.51(m，1H)，3.69(s，4H)，3.04(s，4H)，2.36(s，3H)，2.3(s，3H)，1.65(d，J＝4.0Hz，3H)；LCMS[M+H] ⁺ 491.1.

Example 4, 1- (dodecanoyloxy) ethyl 4- (2- ((2, 4-dimethylphenyl) thio) phenyl) piperazine-1 carboxylic acid (Compound EP-0015-010)

150mg (0.371 mmol) of intermediate EP0015-003-A, 600mg (1.85 mmol) of cesium carbonate and 111mg (0.557 mmol) of lauric acid were dissolved in DMF (5 ml), and the reaction was stirred at room temperature for 3h, and TLC monitoring the starting material was essentially complete. The reaction mixture was extracted twice with 25mL of ethyl acetate after adding 30mL of water, and the organic layer was taken up in Na ₂ SO ₄ Drying and concentrating to obtain crude product. Column chromatography with eluent (petroleum ether: ethyl acetate=2:1, v/v) gave 45mg of white solid, 22% yield, HPLC purity: 97%.

¹ HNMR(400MHz，CDCl ₃ )δ7.36(d，J＝8.0Hz，1H)，7.15(s，1H)，7.10-7.00(m，3H)，6.90-6.86(m，1H)，6.53-6.51(m，1H)，3.67(s，4H)，3.03(s，4H)，2.62-2.55(m，1H)，2.36(s，3H)，2.31(s，3H)，2.36-2.28(m，8H)，1.67-1.61(m，2H)，1.51(d，J＝8.0Hz，3H)，1.34-1.20(m，16H)，0.09-0.85(m，3H)，[M+H] ⁺ 569.2.

Example 5, 1- (hexadecanoyloxy) ethyl 4- (2- ((2, 4-dimethylphenyl) thio) phenyl) piperazine-1-carboxylic acid (Compound EP-0015-011)

In a 50mL single-necked flask, 200mg of intermediate EP0015-003-A (0.49 mmol), 151mg of palmitic acid (0.59 mmol), 479mg of cesium carbonate (1.47 mmol) and 4mL of N, N-dimethylformamide were added, and the mixture was stirred at 40℃for 2 hours. After the reaction, 10mL of water and 15mL of ethyl acetate were added to extract an organic phase, and the organic phase was concentrated under reduced pressure to obtain a crude product, which was subjected to plate separation (petroleum ether: ethyl acetate=20:1, v/v) to obtain 160mg of a yellow oil, yield 51.8%, purity 97%.

¹ H NMR(400MHz，CDCl ₃ )δ7.36(d，J＝7.8Hz，1H)，7.15(s，1H)，7.11-6.99(m，3H)，6.92-6.84(m，2H)，6.52(m，1H)，3.67(m，4H)，3.03(m，4H)，2.49-2.20(m，8H)，1.62(q，J＝7.2Hz，2H)，1.51(d，J＝5.6Hz，3H)，1.24(m，24H)，0.90-0.84(m，3H)；LCMS[M+H] ⁺ 569.2.

Example 6, 1- ((2-propylpentanoyl) oxy) ethyl 4- (2- ((2, 4-dimethylphenyl) thio) phenyl) piperazine-1-carboxylic acid (Compound EP-0015-013)

In a 50mL single-necked flask, 150mg of intermediate EP0015-003-A (0.37 mmol), 81mg of valproic acid (0.56 mmol), 362mg of cesium carbonate (1.11 mmol) and 3mL of N, N-dimethylformamide were added and reacted under stirring at 40℃for 2 hours. After the reaction, 10mL of water and 15mL of ethyl acetate were added to extract an organic phase, and the organic phase was concentrated under reduced pressure to obtain a crude product, which was subjected to plate separation (petroleum ether: ethyl acetate=10:1, v/v) to obtain 100mg of a colorless oil, yield 52.7%, purity 99%.

¹ H NMR(400MHz，CDCl ₃ )δ7.36(d，J＝7.8Hz，1H)，7.15(s，1H)，7.05(m，3H)，6.94-6.85(m，2H)，6.53(m，1H)，3.65(m，4H)，3.02(m，4H)，2.42-2.28(m，7H)，1.66-1.58(m，2H)，1.51(d，J＝5.6Hz，3H)，1.47-1.39(m，2H)，1.32(m，4H)，0.90(t，J＝7.2Hz，6H)；LCMS[M+H] ⁺ 513.2.

Example 7, 1- (2- (1- (aminomethyl) cyclohexyl) acetoxy) ethyl 4- (2- ((2, 4-dimethylphenyl) thio) phenyl) piperazine-1-carboxylic acid trifluoroacetate salt (Compound EP-0015-014 trifluoroacetate salt)

(1) In a 50mL single port flask, 180mg of intermediate EP0015-003-A (0.44 mmol), 179mg of 2- (1- (((tert-butoxycarbonyl) amino) methyl) cyclohexyl) acetic acid (0.66 mmol), 430mg of cesium carbonate (1.32 mmol) and 3mLN, N-dimethylformamide were added and reacted at 40℃for 0.5 hours with stirring. After the reaction, 10mL of water and 15mL of dichloromethane are added to extract an organic phase, the organic phase is decompressed and concentrated to obtain a crude product, the crude product is separated by a pre-preparation plate (petroleum ether: ethyl acetate=10:1, v/v) to obtain 150mg of colorless oily matter, the yield is 52.7%, and LCMS [ M+H-56 ] ] ⁺ 584.2.

(2) In a 50mL single-necked flask, 150mg of the product of the third step, 1.5mL of trifluoroacetic acid and 1.5mL of methylene chloride were added, and the reaction was stirred at room temperature for 0.5 hour. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain 170mg of a colorless oil with a purity of 98%.

¹ H NMR(400MHz，Chloroform-d)δ7.51(m，3H)，7.31(d，J＝7.8Hz，1H)，7.19-7.09(m，3H)，7.03(d，J＝7.7Hz，1H)，6.97(m，1H)，6.83(m，1H)，6.66-6.59(m，1H)，3.73(m，4H)，3.19(m，4H)，2.58(s，2H)，2.36(s，3H)，2.31(s，3H)，1.61-1.38(m，15H)；LCMS[M+H] ⁺ 540.2.

Example 8, 1- ((3- (aminomethyl) -5-methylhexanoyl) oxo) ethyl 4- (2- ((2, 4-dimethylphenyl) thio) phenyl) piperazine-1-carboxylic acid trifluoroacetate (Compound EP-0015-015 trifluoroacetate)

(1) In a 50mL single port flask, 150mg of intermediate EP0015-003-A (0.37 mmol), 145mg of 3- (((t-butyl)Oxycarbonyl) amino) methyl) -5-methylhexanoic acid (0.56 mmol), 362mg cesium carbonate (1.11 mmol) and 3mL N, N-dimethylformamide were reacted with stirring at 40℃for 0.5 h. After the reaction, 10mL of water and 15mL of dichloromethane are added to extract an organic phase, the organic phase is decompressed and concentrated to obtain a crude product, the crude product is separated by a pre-preparation plate (petroleum ether: ethyl acetate=3:1, v/v) to obtain 180mg of colorless oily matter, the yield is 77.4%, and LCMS [ M+H-56 ]] ⁺ 572.2.

(2) In a 50mL single-necked flask, 110mgEP0015-015-A, 1mL of trifluoroacetic acid and 1mL of methylene chloride were added, and the reaction was stirred at room temperature for 0.5 hour. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain 110mg of a colorless oil with a purity of 97%.

¹ H NMR(400MHz，DMSO-d ₆ )δ7.76(m，3H)，7.33(d，J＝8.0Hz，1H)，7.24(m，1H)，7.15-7.06(m，3H)，6.94(m，1H)，6.73(m，1H)，6.44-6.37(m，1H)，3.53(m，4H)，2.97(m，4H)，2.78(m，2H)，2.61-2.51(m，1H)，2.33(s，3H)，2.24(s，3H)，2.12(m，1H)，1.61(m，1H)，1.46(d，J＝5.6Hz，3H)，1.24-1.04(m，3H)，0.88-0.82(m，6H)，LCMS[M+H] ⁺ 528.2.

Test examples

Test drug

EP-X was prepared as described in example 6 of PCT/CN2017/088493, the structure of which is shown below:

each of the other test compounds of the present invention was prepared as described in examples 1-8 above.

Test example 1 microsomal metabolic stability and plasma stability test of test prodrug molecules

Test examples 1-1, microsomal metabolic stability test

Experimental system

(1) Stock solutions of the compounds to be tested and the positive control were prepared at a concentration of 10mM using DMSO as diluent, and then all stock solutions were diluted with 70% acetonitrile to a working concentration of 0.25 mM;

(2) The cofactor used in the test was NADPH regeneration system consisting of 6.5mM NADP,16.5nM G-6-P,3U/mL G-6-P D;

(3) The quencher consists of acetonitrile containing tosyl butyramide and propanol (used as internal standard);

(4) The buffer used in the test was 100mM phosphate buffer containing 3.3mM MgCl ₂ ；

(5) The mixture contained 0.2mg/mL liver microsomal protein and 1 μm test compound/positive control and incubated in 100mM potassium phosphate buffer.

Experimental method

(1) A 0 minute sample was prepared by adding 80 μl aliquots of each incubation mixture to 300 μl of quenching reagent to precipitate the protein. The sample was vortexed and then an aliquot of 20. Mu.L of NADPH regeneration system was added.

(2) The reaction was started by adding 130. Mu.L of NADPH regeneration system to 520. Mu.L of each incubation mixture. The final incubation conditions reached in 650 μl were: 0.2mg/mL microsomal protein, 1. Mu.M test substance/positive control, 1.3mM NADP,3.3mM 6-phosphoglucose, 0.6U/mL 6-phosphoglucose dehydrogenase.

(3) The mixture was incubated with gentle shaking in a 37℃water bath. 100. Mu.L aliquots of each mixture were transferred to clean 96-well plates at 5, 10, 30, 60 minutes, which contained 300. Mu.L of quencher to precipitate the protein, followed by centrifugation (5000 Xg, 10 minutes).

(4) 100. Mu.L of the supernatant was placed in a 96-well assay plate into which 300. Mu.L of ultrapure water was previously added, and then analyzed by LC-MS/MS.

Results

TABLE 1 test Metabolic stability data of Compounds in microsomes

Conclusion(s)

The results shown in the table 1 show that the compound of the invention has longer half-life period in the microsome metabolism process and relatively lower clearance rate, which suggests that the compound of the invention has good microsome metabolism stability, can maintain effective blood concentration in vivo for a longer time, and is obviously better than EP-X.

Test examples 1-2, plasma stability test

Experimental system

(1) Stock solutions of test compounds and positive controls were prepared at a concentration of 10mM using DMSO as diluent. The stock solution of the positive control was then diluted with 50% acetonitrile to a working concentration of 0.2mM, and the stock solution of the test compound was then diluted with 50% acetonitrile to a working concentration of 1 mM.

(2) The quencher consisted of acetonitrile containing tosyl butyramide and propranolol (used as internal standard).

Experimental method

(1) Positive control and test article working solutions (in duplicate) were added to plasma and concentrated to 1 μm and 5 μm, respectively.

(2) A 0 minute sample was prepared by adding 80 μl aliquots of each incubation mixture to 320 μl of quenching reagent to precipitate the protein.

(3) The mixture was incubated in a 37 ℃ water bath with gentle shaking. An 80. Mu.L aliquot of each mixture was transferred to a clean 96-well plate at 30, 60, 120, 240 minutes, which contained 320. Mu.L of quencher to precipitate the protein, followed by centrifugation (4000 Xg, 15 minutes).

(4) 80. Mu.L of the supernatant was placed in a 96-well assay plate with 160. Mu.L of ultrapure water added in advance, and then analyzed by LC-MS/MS.

Results

Table 2 metabolic stability data for test compounds in plasma

Conclusion(s)

From the results shown in Table 2, the compounds of the present invention have a longer plasma half-life, suggesting that the compounds of the present invention can maintain an effective blood concentration in vivo for a longer period of time, which is significantly superior to EP-X.

Test examples 1-3, in vivo Metabolic Properties of the test drug in rats

The purpose is as follows: the pharmacokinetic profile of vortioxetine prodrug compounds of the present invention was examined in comparison with EP-X in rats using LC-MS method.

Chromatographic conditions: the column was a WELCH Xtime-C18 column (50X 2.1mm,3.5 mm); mobile phase: a: aqueous solution (0.1% formic acid); b: acetonitrile (0.1% formic acid); gradient elution (0.10-0.80 min,10% B-95% B, 0.80-1.80min,95% B-95% B, 1.80-1.81min,95% B-10% B, 1.81-3.00min,10% B-10% B;); flow rate: 350 mu L min-1; column temperature: 30 ℃.

Mass spectrometry conditions: positive ion scanning multiple reaction monitoring mode (MRM) using electrospray ion source (ESI). Ion source temperature: 500 ℃; ion ejection voltage: 5000V; atomizer (GS 1): 50psi; assist gas (GS 2): 35psi; collision gas (CAD): high; curtain gas (CUR): 25psi. The ion pairs used for quantitative analysis were m/z 299.3- > 109.2 (vortioxetine), 455.3- > 165.1 (internal standard verapamil), respectively.

Treatment of plasma samples: 20. Mu.L of the plasma sample was taken, placed in a 2.0mL centrifuge tube, and 200. Mu.L of a precipitant solution (acetonitrile solution containing 60.0 ng. Multidot.mL) was added ^-1 Dexamethasone, 5.0 ng.mL ^-1 Verapamil, 30.0 ng.mL of (E) ^-1 Buspirone solution of (a) for 3min by vortexing, centrifuging (12,000 rpm) for 3min, transferring 75 μl of the supernatant to a 96-well plate, adding 75 μl of aqueous solution, mixing well, and taking 10 μl for LC-MS/MS analysis.

Treatment of brain tissue samples: a brain tissue sample (50. Mu.L) was placed in a 2.0mL centrifuge tube, and 500. Mu.L of a precipitant solution (acetonitrile solution containing 60.0 ng. Multidot.mL) was added ^-1 Dexamethasone, 5.0 ng.mL ^-1 Verapamil, 30.0 ng.mL of (E) ^-1 Buspirone solution of (a) for 3min by vortexing, centrifuging (12,000 rpm) for 3min, transferring 75 μl of the supernatant to a 96-well plate, adding 75 μl of aqueous solution, mixing well, and taking 10 μl for LC-MS/MS analysis.

Preparing a test product:

calculating positive control and test compound groups as dosage of vortioxetine with equal dosage according to molecular weight and compound purity of the prodrug, treating the sample by using a spray drying process, then using pure water as a solvent, adding a proper volume of solvent, fully dissolving and uniformly mixing to prepare administration solutions of vortioxetine prodrug groups.

When preparing a test sample, calculating theoretical sample weight according to the designed concentration and the required volume, and calculating the dosage of the vortioxetine prodrug as an equal dosage according to the molecular weight and the purity of the compound in each group of vortioxetine prodrugs. Labeling the prepared medicinal liquid for later use. If the test sample is poorly dissolved or suspended unevenly, appropriate vortexing or ultrasound can be performed.

And (3) test design:

each compound was administered to 18 healthy male rats, 3 animals at each time point, and 6 time points (0.5, 1, 2, 4, 6, 10 hours) at the same dosage of vortioxetine as 10mg/kg, and each group was administered simultaneously in a single intragastric administration. Plasma and whole brain tissue samples were collected at designated time points after dosing. The compound concentrations (vortioxetine prodrug and vortioxetine) in the samples were then determined using API-4000QTRAP LC-MS. Relevant PK parameters were calculated using WinNonLin software and pharmacokinetic parameters were reported as follows: AUC (AUC) _(0-t) 、AUC _(0-∞) 、MRT _(0-t) 、C _max 、T _max 、T _1/2 。

Cerebral blood ratio (B/P) =auc _0-last (brain tissue)/AUC _0-last (plasma)

Results of pharmacokinetic parameters of vortioxetine detected in rat brain tissue and plasma are shown in table 3 for positive control and test compound groups and in rat brain tissue and plasma for prodrugs are shown in table 4.

Results

Table 3 test of pharmacokinetic parameters of vortioxetine of compounds in rats

TABLE 4 pharmacokinetic parameters of prodrugs of compounds tested in rats

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Conclusion(s)

From the results shown in tables 3 to 4, it is evident that the metabolite vortioxetine of the compound EP-0015-010 of the present invention has a cerebral blood ratio (B/P) of 50.47, the metabolite vortioxetine of EP-X has a B/P of 33.54, and the metabolite vortioxetine of EP-0015-010 has a significantly increased cerebral blood ratio (B/P) relative to EP-X, suggesting that the compound of the present invention has a stronger targeted administration effect to brain tissues and significantly increases the exposure of the active metabolite vortioxetine in the brain.

In addition, according to tables 3-4, it can be found that the vortioxetine prodrug EP-0015-010 can maintain a certain prodrug blood concentration after entering blood and brain while maintaining quick release of vortioxetine as an active product, and the prodrug EP-0015-010 is provided with reasonable prodrug clearance rate and release rate of vortioxetine as an active metabolite; whereas the exposure of EP-X not only metabolite vortioxetine in brain tissue was significantly lower than that of EP-0015-010 (AUC _0-last 10800vs 6640), which is also difficult to detect in plasma and brain tissue itself, suggests that clearance is too fast to maintain effective vortioxetine blood levels for extended periods of time.

Test example 2 in vivo pharmacokinetic testing of prodrug molecules in rats

The purpose is as follows: the pharmacokinetic profile of vortioxetine prodrug compound in rats was examined using the LC-MS method.

Chromatographic conditions: the column was a WELCH Xtime-C18 column (50X 2.1mm,3.5 μm); mobile phase: a: aqueous solution (0.1% formic acid); b: acetonitrile (0.1% formic acid); gradient elution (0.10-0.80 min,10% B-95% B, 0.80-1.80min,95% B-95% B, 1.80-1.81min,95% B-10% B, 1.81-3.00min, 1)0% B-10% B; ) The method comprises the steps of carrying out a first treatment on the surface of the Flow rate: 350 mu L min ^-1 The method comprises the steps of carrying out a first treatment on the surface of the Column temperature: 30 ℃.

Treatment of brain tissue samples: a brain tissue sample (50. Mu.L) was placed in a 2.0mL centrifuge tube, and 500. Mu.L of a precipitant solution (acetonitrile solution containing 60.0 ng. Multidot.mL) was added ^-1 Dexamethasone, 5.0 ng.mL ^-1 Verapamil, 30.0 ng.nL of (E) ^-1 Buspirone solution of (a) for 3min by vortexing, centrifuging (12,000 rpm) for 3min, transferring 75 μl of the supernatant to a 96-well plate, adding 75 μl of aqueous solution, mixing well, and taking 10 μl for LC-MS/MS analysis.

Preparing a test product:

vortioxetine group: 28mg of vortioxetine compound (self-made) is precisely weighed, 5% DMSO solution and 95% (0.5% CMC-Na solution) are used as solvents, a proper volume of solvents are added, and the solvents are fully dissolved and uniformly mixed to obtain a complete suspension of 0.5mg/mL, thus obtaining vortioxetine administration solution.

Vortioxetine prodrugs each group: EP-0015-003, EP-0015-004, EP-0015-006, EP-0015-010, EP-0015-011, EP-0015-013, EP-0015-014 and EP-0015-015 are calculated as the dosage of vortioxetine with equal dosage according to the molecular weight and the compound purity of each prodrug, 5% DMSO solution +95% (0.5% CMC-Na solution) is used as a solvent, and a proper volume of solvent is added to fully dissolve and mix the mixture to prepare each group of administration solutions of vortioxetine prodrugs.

And (3) test design:

each compound was administered to 18 healthy male rats, 3 animals at each time point, and 6 time points (0.5, 1, 2, 4, 6, 10 hours) at the same dosage of vortioxetine of 1mg/kg, and each group was administered simultaneously in a single intragastric administration. Plasma and whole brain tissue samples were collected at designated time points after dosing. The compound concentrations (vortioxetine prodrug and vortioxetine) in the samples were then determined using API-4000 QTRAPLC-MS. Relevant PK parameters were calculated using WinNonLin software and pharmacokinetic parameters were reported as follows: AUC (AUC) _(0-t) 、AUC _(0-∞) 、MRT _(0-t) 、C _max 、T _max 、T _1/2 、F％。

Relative bioavailability calculation (Relative Bioavailability (%)) =auc _{0-last (vortioxetine)} (prodrug)/AUC _{0-last (vortioxetine)} ) (vortioxetine)

Cerebral blood ratio (B/P) =auc _0-last (brain tissue)/AUC _0-last (plasma)

The results of the pharmacokinetic parameters of vortioxetine and vortioxetine prodrug compounds detected in rat brain tissue are shown in table 5, the results of the pharmacokinetic parameters of vortioxetine and vortioxetine prodrug compounds detected in rat plasma are shown in table 6, the results of the ratio of vortioxetine AUC (cerebral blood ratio, B/P) of vortioxetine and vortioxetine prodrug compounds in rat brain tissue to vortioxetine AUC (cerebral blood ratio, B/P) are shown in fig. 1, the total AUC exposure of vortioxetine and vortioxetine prodrug compounds is shown in fig. 2, and the relative bioavailability comparisons of vortioxetine and vortioxetine prodrug compounds are shown in fig. 3.

Results

Table 5 pharmacokinetic parameters of vortioxetine and vortioxetine prodrug compounds in rat brain

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Table 6 pharmacokinetic parameters of vortioxetine and vortioxetine prodrug compounds in rat plasma

Conclusion(s)

After single gastric administration of vortioxetine and vortioxetine prodrug compounds in male SD rats, the relative bioavailability (relative to vortioxetine) of the metabolites vortioxetine of EP-0015-003, EP-0015-010, EP-0015-014 and EP-0015-015, and particularly the relative bioavailability of EP-0015-015, is improved significantly in brain tissue of rats compared with vortioxetine, and the compound of the invention has the therapeutic potential of significantly reducing the administration dose of vortioxetine, which is more beneficial for reducing the related adverse effects of drugs, such as reducing the incidence of gastrointestinal side effects.

Compared with vortioxetine, the present invention has significantly improved AUC cerebral blood ratio (B/P) of vortioxetine prodrug compound, such as blood cerebral ratio of EP-0015-003, EP-0015-004, EP-0015-006, EP-0015-010, EP-0015-011, EP-0015-014, EP-0015-015, which has significantly changed compared with blood cerebral ratio of vortioxetine, suggesting that there is stronger targeted administration effect of brain tissue (see FIG. 1), thus the brain exposure (AUC _0-last ) Also higher (see fig. 2), where the same dose of vortioxetine acts on the central nervous system, the prodrugs of the present invention can reduce the dose administered and thus also reduce the effect of vortioxetine on the peripheral nervous system. It can also be seen from figure 2 that the vast majority of vortioxetine prodrugs provided by the present invention, such as EP-0015-003, EP-0015-004, EP-0015-010, EP-0015-014, EP-0015-015, have a total AUC that is substantially the same as or greater than vortioxetine.

In addition, as shown in fig. 4 and fig. 5, compared with vortioxetine, the vortioxetine prodrug provided by the invention has a flatter drug-time curve in blood plasma and brain, which suggests that a relatively stable drug treatment concentration can be maintained in a human body, and has good pharmacokinetic properties.

Test example 3 in vivo pharmacokinetic testing of prodrug molecule cynomolgus monkey

The purpose is as follows: LC-MS method was used to examine the pharmacokinetic profile of vortioxetine prodrug compounds in cynomolgus monkeys.

Chromatographic conditions: the chromatographic column is ACQUITY UPLC Protein BEH C41.7 μm 2.1X150 mm; mobile phase: a:2mM HCOONH ₄ in water/ACN (v: v, 95: 5); b:2mM HCOONH4in ACN/water (v: v, 95: 5); gradient elution (0.10-0.30 min,15% -40% B, 0.30-1.40min,40% -60% B, 1.40-1.60min,60% -90% B, 1.60-2.00min,90% B,2.00-2.20min,90% B-15% B); flow rate: 700 mu L min-1; column temperature: 55 ℃.

Mass spectrometry conditions: the electrospray ion source (ESI) was used and the reaction detection scanning mode (SRM) was selected. Ion source temperature: 500 ℃; ion ejection voltage: 5000V; atomizer (GS 1): 50psi; assist gas (GS 2): 35psi; collision gas (CAD): high; curtain gas (CUR): 25psi. The ion pairs used for quantitative analysis were m/z 300.4.fwdarw.150.2 (vortioxetine), 455.2.fwdarw.164.9 (internal standard verapamil), respectively.

Treatment of plasma samples: aliquots of 20 μl of unknown sample, calibration standard, quality control, dilution quality control, single blank and double blank samples were added to 96-well plates; each sample (except for the double blank) was quenched with 200 μl LIS1 (double blank with 200 μl MeOH), and the mixture was then vortexed at 800rpm for 10 minutes, then centrifuged at 3220g for 15 minutes (4000 rpm), 4 ℃. 2. Mu.L of supernatant was injected for LC-MS/MS analysis;

treatment of cerebrospinal fluid samples: plasma and ethanol (v; v; v 1; 1) were added to all cerebrospinal fluid samples; aliquots of 20 μl of unknown sample, calibration standard, quality control, dilution quality control, single blank and double blank samples were added to 96-well plates; each sample (except for the double blank) was quenched with 200 μl of IS1 (double blank with 200 μl of MeOH), and the mixture was then vortexed at 800rpm for 10 minutes, then centrifuged at 3220g for 15 minutes (4000 rpm), 4 ℃. mu.L of supernatant was injected for LC-MS/MS analysis.

Preparing a test product:

vortioxetine group: precisely weighing vortioxetine compound, adding a proper volume of solvent by using 90% PEG400/10% water as the solvent, fully dissolving and uniformly mixing to obtain a complete suspension of 2mg/mL, wherein the complete suspension is vortioxetine administration solution.

EP-0015-010 group: the EP-0015-010 compound is precisely weighed, 90% PEG400/10% water is used as a solvent, a proper volume of solvent is added, and the mixture is fully dissolved and uniformly mixed to obtain 3.82mg/mL of complete suspension, namely an EP-0015-010 dosing solution (equivalent to 2mg/mL of vortioxetine).

When preparing a test sample, calculating theoretical sample weight according to the designed concentration and the required volume, and calculating the dosage of vortioxetine with the equivalent dosage of 3mg/kg according to the molecular weight and the purity of the compound in each group of vortioxetine prodrug. Labeling the prepared medicinal liquid for later use. If the test sample is poorly dissolved or suspended unevenly, appropriate vortexing or ultrasound can be performed.

And (3) test design:

taking 2 healthy male and female cynomolgus monkeys respectively, wherein the age is more than or equal to 2 years old, and the weight is more than or equal to 2.5kg. Experiments were carried out on 4 cynomolgus monkeys, and the experiments were carried out at intervals of one week before and after each other for metabolism of the drug, and the experiments were carried out according to the administration dose of vortioxetine at an equivalent dose of 3mg/kg, and blood plasma and cerebrospinal fluid samples were collected at 6 time points (0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours) respectively. The concentration of the compound (vortioxetine and vortioxetine prodrug) in the sample was then determined using the achquityupllcms-MS. Relevant PK parameters were calculated using WinNonLin software and pharmacokinetic parameters were reported as follows: AUC (AUC) _(0-t) 、AUC _(0-∞) 、MRT _(0-t) 、C _max 、T _max 、T _1/2 、F％。

Relative bioavailability calculation (Relative Bioavailability (%)) =auc0-last _{(Wo Xixi tin)} (prodrug)/AUC 0-last _{(vortioxetine)} (vortioxetine)

Cerebral blood ratio (B/P) =auc _0-last (cerebrospinal fluid)/AUC _0-last (plasma)

The results of the pharmacokinetic parameters of vortioxetine detected by vortioxetine prodrug compounds in cynomolgus monkey plasma and cerebrospinal fluid are shown below 7; the results of total AUC exposure of vortioxetine and vortioxetine prodrug compounds in cynomolgus cerebrospinal fluid and plasma are shown in figure 6.

Results

TABLE 7 pharmacokinetic parameters of vortioxetine and vortioxetine prodrug compounds in cynomolgus monkey plasma and cerebrospinal fluid

Remarks: "NA" indicates undetected.

Conclusion(s)

From the above results, it is clear that, compared with vortioxetine, the vortioxetine prodrug provided by the present invention has a total AUC significantly higher than that of vortioxetine, and the vortioxetine prodrug metabolites vortioxetine and vortioxetine are substantially equal (see fig. 6 in particular); in addition, the B/P ratio of vortioxetine drug concentration generated after the vortioxetine prodrug is metabolized is obviously higher than that of vortioxetine, which indicates that the prodrug provided by the invention can be targeted for brain administration. In addition, the concentration of the free vortioxetine produced by metabolism of the prodrug compound in cerebrospinal fluid is far away from the concentration of the free drug of Gao Yuwo vortioxetine in cerebrospinal fluid, which indicates that the prodrug can obviously improve the effective therapeutic concentration of vortioxetine, so that the prodrug provided by the invention can improve the anti-depression effect, and meanwhile, the administration dosage can be reduced on the premise of maintaining the same drug effect, and further adverse reaction is reduced.

The above embodiments are provided to illustrate the technical concept and features of the present invention and are intended to enable those skilled in the art to understand the content of the present invention and implement the same, and are not intended to limit the scope of the present invention. All equivalent changes or modifications made in accordance with the spirit of the present invention should be construed to be included in the scope of the present invention.

Claims

1. A compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof,

wherein:

x-is a single bond "-" or- (CH) ₂ )n ₁ -any one of the following;

R ₁ and R is ₂ Each independently selected from C _1-3 Or hydrogen;

R ₄ and R is ₅ Each independently selected from hydrogen, amino, C optionally substituted with amino _6-15 Or C optionally substituted by amino _1-5 Any one of the alkyl groups of (a);

n ₁ is any integer between 1 and 3;

a and b are each independently any integer between 0 and 2.

2. The compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof according to claim 1,

R ₁ and/or R ₂ In the above, the C _1-3 The alkyl of (2) is selected from any one of methyl, ethyl or propyl；

And/or R ₃ In the above, the C _6-10 Is selected from any one of phenyl or naphthyl;

And/or R ₄ And R is ₅ In the above, the C _6-15 The alkyl group of (2) is selected from C _6-15 A linear alkyl group of (a);

and/or R ₄ And R is ₅ In said C optionally substituted with amino _1-5 Is selected from any one of methyl, ethyl, propyl, butyl or amyl optionally substituted by amino;

and/or, the n ₁ 1 is shown in the specification;

and/or, each of said a and b is independently 1.

3. The compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof according to claim 2,

R ₁ and/or R ₂ In the above, the C _1-3 Is any one of methyl or ethyl;

and/or R ₃ In the above, the C _6-10 Aryl of (2) is phenyl;

and/or R ₄ And R is ₅ In the above, the C _6-15 Is selected from C _9-15 A linear alkyl group of (a);

and/or R ₄ And R is ₅ In said C optionally substituted with amino _1-5 Is any one of methyl, ethyl, propyl or butyl optionally substituted with amino.

4. The compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof according to claim 3, wherein R ₄ And R is ₅ In said C optionally substituted with amino _1-5 Is any one of methyl, ethyl or propyl optionally substituted with amino.

5. The compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein R ₃ Is phenyl orA naphthyl group.

6. The compound of formula (a) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein R ₁ And R is ₂ Each independently selected from any one of hydrogen, methyl or ethyl.

7. The compound of formula (a) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein R ₁ And R is ₂ Each independently selected from hydrogen or methyl.

8. The compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof according to claim 6, wherein R ₃ Is phenyl or naphthyl.

9. The compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein R ₄ And R is ₅ Each independently is hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, -CH ₂ NH ₂ ，-CH ₂ CH ₂ NH ₂ ，-CH ₂ CH ₂ CH ₂ NH ₂ ，-(CH ₂ ) ₈ CH ₃ ，-(CH ₂ ) ₉ CH ₃ ，-(CH ₂ ) ₁₂ CH ₃ Or- (CH) ₂ ) ₁₃ CH ₃ Any one of the following.

10. The compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein R ₄ And R is ₅ Each independently is hydrogen, methyl, n-propyl, isopropyl, -CH ₂ CH ₂ NH ₂ ，-(CH ₂ ) ₈ CH ₃ ，-(CH ₂ ) ₉ CH ₃ ，-(CH ₂ ) ₁₂ CH ₃ Or- (CH) ₂ ) ₁₃ CH ₃ Any one of the following.

11. The compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof according to claim 1,

X-is a single bond "-" or- (CH) ₂ )n ₁ -any one of the following;

R ₃ is selected from the group consisting of phenyl groups,any one of them;

R ₄ and R is ₅ Each independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, -CH ₂ NH ₂ ，-(CH ₂ ) ₈ CH ₃ ，-(CH ₂ ) ₉ CH ₃ ，-(CH ₂ ) ₁₂ CH ₃ Or- (CH) ₂ ) ₁₃ CH ₃ Any one of them;

a is 1, b is 1;

n ₁ is any integer between 1 and 3.

12. The compound of formula (a) or a pharmaceutically acceptable salt thereof as claimed in claim 11, wherein C _1-3 The alkyl group of (a) is selected from any one of methyl and ethyl.

13. The compound of formula (a) or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of formula (a) is selected from any one of the following compounds:

14. the compound of formula (a) or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of formula (a) is selected from the group consisting of:

15. a compound of formula (a) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, hydrobromide, hydroiodide, nitrate, sulphate, bisulphate, phosphate, acid phosphate, acetate, lactate, citrate, tartrate, maleate, fumarate, mesylate, trifluoroacetate, gluconate, saccharate, benzoate, ethanesulphonate, benzenesulfonate or p-toluenesulfonate.

16. A compound of general formula (a), a stereoisomer thereof, a tautomer thereof or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein the pharmaceutically acceptable salt is selected from the group consisting of hydrobromide and trifluoroacetate.

17. A process for preparing a compound of general formula (a) or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 14, which comprises:

wherein:

-X-，R ₁ ，R ₂ and R is ₃ Each independently as defined in any one of claims 1 to 14.

18. The method of claim 17, wherein the method of preparing the compound of formula (III) comprises:

wherein:

R ₁ and R is ₂ Each independently as defined in any one of claims 1 to 14.

19. A pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (a) according to any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

20. Use of a compound of general formula (a) or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 14 or a pharmaceutical composition as claimed in claim 19 in the manufacture of a medicament for the treatment of neuropsychiatric diseases.

21. The use according to claim 20, wherein the neuropsychiatric disorder is selected from any one or more of schizophrenia, depression, anxiety, sleep disorders, cognitive disorders, neurodegenerative disorders, bipolar disorders, post-traumatic stress syndrome, addictive disorders, withdrawal syndrome or attention deficit.

22. The use according to claim 20, wherein the neuropsychiatric disorder is selected from any one or more of depression, cognitive disorders, anxiety, schizophrenia, sleep disorders, neurodegenerative disorders or bipolar disorders.

23. The use according to claim 20, wherein the neuropsychiatric disease is selected from depression.

24. The use according to any one of claims 21 to 23, wherein the depression is selected from the group consisting of mild depression, moderate depression, major depression, depression accompanied by other mental disorders, and recurrent depression; the other mental disorder is selected from one or more of sleep disorder, cognitive disorder, anxiety disorder, obsessive-compulsive disorder, affective disorder or personality disorder.

25. The use according to claim 24, wherein the mental disorder is selected from cognitive disorders, anxiety disorders or sleep disorders.