JP2021507889A - New use of pyrazolopiperidine derivatives - Google Patents

Abstract

The present invention relates to the use of specific pyrazoropiperidine derivatives in the treatment of diseases and conditions associated with toll-like receptor interactions. More specifically, the treatment relates to the treatment of NPSLE.

Description

The present invention relates to the use of certain pyrazole derivatives in the treatment of diseases and conditions that may be associated with Toll-like receptor interactions. More specifically, the procedure relates to the treatment of neuropsychiatric systemic lupus erythematosus (NPSLE), including central nervous system (CNS) lupus.

U.S. Pat. Nos. 9,126,999 B2 and WO2018 / 047081 pamphlet describe the synthesis and use of certain pyrazolo-piperidin derivatives. The compounds are known to antagonize Toll-like receptors 7 and 8 (TLR7, TLR8) and have been shown to be useful in the treatment of SLE and lupus nephritis. However, US Pat. No. 9,126,999 B2 and WO 2018/047081 pamphlet do not describe the treatment of NPSLE.

The underlying pathophysiological mechanism of NPSLE, including CNS lupus, remains largely unknown.

However, several pathogenic pathways have been identified, including type I interferon-dependent pathways (see Bialas et al. Nature. 2017, Jun 22: 546 (7659): 539-543). Currently, there is considered no treatment available for the treatment of NPSLE, including CNS lupus, and there is a high medical need for each treatment.

The present invention is a novel method for treating and / or preventing an NPSL in a subject in need of such treatment, in an effective amount of a compound of formula (I) or compound of formula (II); or Provided are methods comprising administering a TLR7 / TLR8 antagonist, such as the pharmaceutically acceptable salt, to a patient suffering from NPSLE.

Efficacy of compounds of formula (Ib) for systemic disease parameters in the NZBW / F1 model. Efficacy of compounds of formula (IIc) for systemic and neuropathological disease parameters in the NZBW / F1 model.

In a first embodiment, the invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of NPSLE. The compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

(During the ceremony
R _{1 is a C 1 to} C ₄ alkyl that is optionally substituted at least once with hydrogen or halogen;
R ₂ and R ₃ are independent of each other, hydrogen or C _{1 to} C ₆ alkoxy;
R ₄ is hydrogen or C _{1 to} C ₆ alkyl;
R ₅ is hydrogen or C _{1 to} C ₆ alkyl)
Represented by.

In the second embodiment, the present invention describes the formula (I) for use according to the first embodiment (in the formula,
R _{1 is a C 1 to} C ₄ alkyl that is optionally substituted with halogen at least once;
R ₂ and R ₃ are independent of each other and are C _{1 to} C ₆ alkoxy;
R ₄ is hydrogen or C _{1 to} C ₆ alkyl;
R ₅ is hydrogen or methyl)
To provide a compound of or a pharmaceutically acceptable salt thereof.

In a third embodiment, the present invention describes the formula (I) for use as described in the first embodiment.
R _{1 is a C 1 to} C ₄ alkyl substituted with halogen at least once;
R ₂ and R ₃ are independent of each other and are C _{1 to} C ₆ alkoxy;
R ₄ is hydrogen or C _{1 to} C ₆ alkyl;
R ₅ is hydrogen)
To provide a compound of or a pharmaceutically acceptable salt thereof.

In a fourth embodiment, the present invention describes the formula (I) for use as described in the first embodiment.
R ₁ is trifluoromethyl or difluoromethyl;
R ₂ and R ₃ are both methoxy;
R ₄ is hydrogen;
R ₅ is hydrogen or methyl)
To provide a compound of or a pharmaceutically acceptable salt thereof.

In a fifth embodiment, the present invention provides a compound of formula (I) for use according to the first embodiment or a pharmaceutically acceptable salt thereof, wherein said compound is of formula ( Ia)

Or a pharmaceutically acceptable salt thereof.

In a sixth embodiment, the present invention provides a compound of formula (I) for use according to the first embodiment or a pharmaceutically acceptable salt thereof, wherein the compound is of formula ( Ib)

Or a pharmaceutically acceptable salt thereof.

In a seventh embodiment, the invention provides a compound of formula (I) for use according to the first embodiment or a pharmaceutically acceptable salt thereof, wherein the compound is (4). -((5S, 7R) -5- (3,4-dimethoxyphenyl) -7- (trifluoromethyl) -4,5,6,7-tetrahydropyrazolo [1,5-a] pyrimidin-2-yl ) Phenyl) (Piperazine-1-yl) Metanon.

In Embodiment 8, the present invention provides a compound of formula (II) or a pharmaceutically acceptable salt thereof for use in the treatment and / or prevention of NPSLE. The compound of formula (II) or a pharmaceutically acceptable salt thereof has the following structure:

(During the ceremony
L is -CH _2- or -CH ₂ CH _2- ;
R ¹ is -NHC (= O) R ⁶ , -NHC (= O) (CH ₂ ) _n R ⁶ , -NHC (= O) (CH ₂ ) _m NHR ⁵ , -NHC (= O) (CH _{2) ^{) m n (R 5) 2}} , -NHC (= O) (CHR 7) m NHR 5, -NHC (= O) (CH 2) m NH 2, -NHC (= O) (CH 2) n OR 7 , -NHC (= O) OR ⁷ , -NHC (= O) (CHR ⁷ ) _n R ⁶ , -NHC (= O) (CHR ⁷ ) _n N (R ⁸ ) ₂ , -NHC (= O) (CHR) ⁷ ) _n NHR ⁸ , -NHC (= O) (CH ₂ ) _n N (CD ₃ ) ₂ , -NR ⁷ C (= O) R ⁵ , -NR ⁷ C (= O) (CH ₂ ) _n R ⁵ , -NR ⁷ C (= O) OR ⁵ , -NHS (= O) ₂ R ⁵ , -NHC (= O) (CH ₂ ) _n NR ⁷ C (= O) R ⁵ or -NHC (= O) ( CH ₂ ) _n NR ⁷ S (= O) ₂ R ⁵ ;
R ² is, _H, be a C 1 -C ₆ alkyl or _C 1 -C ₆ haloalkyl;
R ³ is H, C _{1 to} C ₆ alkyl or -CD ₃ ;
R ⁴ is H, NH ₂ , C _{1 to} C ₆ alkyl or halo;
Each R ⁵ is independently selected from C _1- C ₆ alkyl, -CD ₃ and- (CH ₂ ) _n OR ⁷ ;
R ⁶ is _{independent of C 3 to} C ₆ cycloalkyl, or N, NH, N (C _{1 to} C ₆ alkyl) and O (unsubstituted or substituted with one or two R ⁹ groups). It is a 4- to 6-membered heterocycloalkyl having 1-2 ring members selected;
Each R ⁷ is independently selected from H and C _1- C _{6 alkyl;}
Each R ⁸ is independently selected from C _{1 to} C ₆ haloalkyl,-(C (R ⁷ ) ₂ ) _n OR ⁷ _{, and C 1 to} C ₆ alkyl substituted with 1 to 3 -OH;
Each ^{R 9} is _{independently} selected C 1 -C ₆ alkyl, hydroxyl, halo, and _C 1 -C ₆ alkyl substituted with 1 to 3 -OH;
n is 1, 2, 3, 4, 5 or 6;
m is 1, 2, 3, 4, 5 or 6)
Represented by.

In the ninth embodiment, the present invention is based on the formula (IIa):

R ² is, _H, be a C 1 -C ₆ alkyl or _C 1 -C ₆ haloalkyl;
R ³ is H, C _{1 to} C ₆ alkyl or -CD ₃ ;
R ⁴ is H, NH ₂ , C _{1 to} C ₆ alkyl or halo;
R ⁶ is _{independent of C 3 to} C ₆ cycloalkyl, or N, NH, N (C _{1 to} C ₆ alkyl) and O (unsubstituted or substituted with one or two R ⁹ groups). It is a 4- to 6-membered heterocycloalkyl having 1-2 ring members selected;
Each ^{R 9} is _{independently,} C 1 -C ₆ alkyl is selected hydroxyl, halo, and _C 1 -C ₆ alkyl substituted with 1 to 3 -OH)
A compound of formula (II) for use according to embodiment 8, which is a compound of the above, or a pharmaceutically acceptable salt thereof.

In the tenth embodiment, the present invention
R ² is C _{1 to} C ₄ alkyl;
R ³ is a C _{1 to} C ₄ alkyl;
R ⁴ is _located at C 1 -C ₄ alkyl or halo;
9. The ninth embodiment, wherein R ⁶ is a 4- to 6-membered heterocycloalkyl having 1-2 ring members that are independently selected from N, NH, N (C _{1 to} C _{6 alkyl) and O.} Provided are compounds of formula (IIa) for use, or pharmaceutically acceptable salts thereof.

In the eleventh embodiment, the present invention is based on the formula (IIb):

(R ² is C _{1 to} C ₄ alkyl;
R ³ is a C _{1 to} C ₄ alkyl;
R ⁴ is C _{1 to} C ₄ alkyl)
A compound of formula (IIa) for use according to embodiment 9 or 10, or a pharmaceutically acceptable salt thereof, which is a compound of.

In embodiment 12, the invention provides a compound of formula (II), (IIa) or (IIb) for use according to any one of embodiments 8-11, or a pharmaceutically acceptable salt thereof. Here, the compound is (S) -N- (4-((5- (1,6-dimethyl-1H-pyrazolo [3,4-b] pyridin-4-yl) -3-methyl-4). , 5,6,7-Tetrahydro-1H-pyrazolo [4,3-c] pyridin-1-yl) methyl) bicyclo [2.2.2] octane-1-yl) morpholin-3-carboxamide; or its pharmacy A reasonably acceptable salt. This compound also has formula (IIc) :.

Represented by.

In embodiment 12a, the invention provides a compound of formula (IIc) in free form for use according to any one of embodiments 8-11.

In embodiment 12b, the invention provides a compound of formula (IIc) in hydrated form for use according to any one of embodiments 8-11.

In embodiment 12c, the present invention describes (S) -N- (4-((5- (1,6-dimethyl-1H-pyrazolo]) for use according to any one of embodiments 8-11. 3,4-b] Pyridine-4-yl) -3-methyl-4,5,6,7-tetrahydro-1H-pyrazolo [4,3-c] pyridin-1-yl) methyl) bicyclo [2.2 .2] Provide a crystalline form of a hydrate of octane-1-yl) morpholine-3-carboxamide. In a further aspect of this embodiment, the present invention describes (S) -N- (4-((5- (1,6-dimethyl-1H)) for use according to any one of embodiments 8-11. -Pyrazolo [3,4-b] Pyridine-4-yl) -3-Methyl-4,5,6,7-Tetrahydro-1H-Pyrazolo [4,3-c] Pyridine-1-yl) Methyl) Bicyclo [ 2.2.2] Provided is a crystalline form of a heptahydrate of octane-1-yl) morpholine-3-carboxamide. Such crystalline morphology is incorporated herein by reference in the patent application PCT / CN2018 / 087448 (agent reference number: PAT058103-WO-PCT) filed May 18, 2018. Have been described.

As used herein, neuropsychiatric lupus erythematosus (NPSL) suffers from systemic lupus erythematosus (SLE), including neuropathy in the central nervous system (CNS) symptoms (CNS lupus) and peripheral nervous system (PNS). Refers to the various neurological and / or behavioral clinical manifestations of the patient. Lupus neuropsychiatric symptoms range from mild to severe (see Kivitity et al. BMC Med. 2015 Mar 4; 13:43), affecting up to 75% of patients with SLE. The effects of neuropsychiatric symptoms on disease severity, quality of life, and prognosis are considered to be one of the leading causes of morbidity and mortality in patients with SLE.

The present invention relates to the treatment of NPSLE patients, typically by inhibition of Toll-like receptors (TLRs), and may include inhibition of TLR7 in particular. Importantly, the Applicant does not want to be bound by mechanical considerations, so the present invention is in any one of embodiments 1-12 for use in the treatment and / or prevention of NPSLE. The compound of formula (I) or the compound of formula (II) described, or a pharmaceutically acceptable salt thereof.

In a further embodiment, the invention is a compound of formula (I) or a compound of formula (II) or a compound according to any one of embodiments 1-12 for use in the treatment and / or prevention of NPSLE. Provided is a pharmaceutical composition comprising; or a pharmaceutically acceptable salt thereof, together with one or more pharmaceutically acceptable carriers.

In yet another embodiment, the invention is a method for treating and / or preventing NPSLE in a patient in need thereof, the effective amount of which is described in any one of embodiments 1-7. Provided are a method comprising administering a compound of formula (I) as described; or a pharmaceutically acceptable salt thereof.

In yet another embodiment, the invention is a method for treating and / or preventing NPSLE in a patient in need thereof, the effective amount of which is described in any one of embodiments 8-12. Provided are a method comprising administering a compound of formula (II) as it is; or a pharmaceutically acceptable salt thereof.

In yet another embodiment, the invention is a method for treating and / or preventing NPSLE in a patient in need thereof, wherein an effective amount is described in any one of embodiments 1-7. Provided are methods comprising administering a pharmaceutical composition comprising a compound of formula (I); or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers.

In yet another embodiment, the invention is a method for treating and / or preventing NPSLE in a patient in need thereof, wherein an effective amount is described in any one of embodiments 8-12. Provided are methods comprising administering a pharmaceutical composition comprising a compound of formula (II); or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers.

In yet another embodiment, the present invention is a compound represented by the formula (I) according to any one of embodiments 1 to 7 in the production of a drug for treating and / or preventing NPSLE, or a compound represented by the formula (I). The use of the pharmaceutically acceptable salt is provided.

In yet another embodiment, the present invention is a compound of formula (II) according to embodiments 7-12, or pharmaceutically acceptable thereof, in the manufacture of a drug for treating and / or preventing NPSLE. Provide the use of salt that can.

To interpret the specification, the following definitions apply, and whenever appropriate, the terms used in the singular form include more than one, and vice versa.

As used herein, the term "alkyl" refers to a fully saturated, branched or non-branched chain hydrocarbon moiety having up to 20 carbon atoms. Unless otherwise indicated, alkyl refers to a hydrocarbon moiety having 1 to 16 carbon atoms, 1 to 10 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. Representative examples of alkyl are, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-. Hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, n-decyl and the like can be mentioned.

As used herein, the term "alkoxy" refers to alkyl-O-, where alkyl is defined above herein. Representative examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, tert-butoxy, pentyloxy, hexyloxy and the like. Typically, the alkoxy group has about 1 to 6 carbons, more preferably about 1 to 4 carbons.

As used herein, the term halogen refers to a fluoro, chloro, bromo or iodo group or residue. The halogen can preferably be fluoro.

As used herein, the term alkyl, which is optionally substituted one or more times with halogen, refers to, for example, methyl, ethyl, propyl, tert-butyl, monofluoromethyl, difluoromethyl, trifluoromethyl, tri. It can refer to fluoroethyl, tetrafluoroethyl or pentafluoroethyl.

Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids.

Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases.

The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effects and properties of the compounds of the invention and is typically not biologically or otherwise inconvenient. In many cases, the compounds of the present invention are capable of forming acidic and / or basic salts in the presence of amino and / or carboxyl groups or similar groups.

Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids, such as acetate, asparagate, benzoate, besilate, bromide / hydrobromide, bicarbonate / carbonate. Salt, Bisulfate / Sulfate, Camper Sulfate, Chloride / Hydrochloride, Chlortheophyllone, Citrate, Ethandisulfonate, Fumalate, Gluceptate, Gluconate, Glucronate Salt, horse urate, hydroiodide / iodide, isethionate, lactate, lactobionate, lauryl sulfate, malate, maleate, malonate, mandelate, mesylate, Methylsulfate, naphthoate, napsylate, nicotinate, nitrate, octadecanoate, oleate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / diphosphate Hydrogen salts, polygalacturonates, propionates, stearate, succinates, sulfosalicylates, tartrates, tosylates and trifluoroacetates.

Examples of the inorganic acid from which the salt can be derived include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like.

Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, etc. Examples thereof include ethanesulfonic acid, toluenesulfonic acid and sulfosalicylic acid. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases.

Examples of the inorganic base from which the salt can be derived include ammonium salts and metals of groups I to XII in the periodic table. In certain embodiments, the salts are derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper; particularly suitable salts include ammonium salts, potassium salts, sodium salts, calcium salts and Examples include magnesium salts.

Examples of the organic base from which the salt can be derived include primary, secondary and tertiary amines, substituted amines containing naturally substituted amines, cyclic amines, basic ion exchange resins and the like. Specific organic amines include isopropylamine, benzathine, corinate, diethanolamine, diethylamine, lysine, meglumine, piperazine and tromethamine.

The pharmaceutically acceptable salts of the present invention can be synthesized from basic or acidic moieties by conventional chemical methods. In general, such salts allow the free acid form of these compounds to react with a stoichiometrically suitable base (Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate, etc.). It can be prepared by or by reacting the free base form of these compounds with a stoichiometric amount of the appropriate acid. Such reactions are typically carried out in water or organic solvents, or in mixtures of both. In general, it is desirable to use a non-aqueous medium such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile where feasible. A list of more suitable salts can be found, for example, in "Remington's Pharmaceutical Sciences", 20th ed. , Mac Publishing Company, Easton, Pa. , (1985); and "Handbook of Physical Salts: Properties, Selection, and Use" by Stahl and Wermus (Wiley-VCH, Weinheim, Germany, 2002).

As used herein, the term "free form" refers to the compound itself without salt formation.

As used herein, the term "pharmaceutically acceptable carrier" is known to those of skill in the art (eg, Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company, 1990, pp. 1289). -1329), any solvent, dispersion medium, coating, surfactant, antioxidant, preservative (eg, antibacterial, antifungal), isotonic, absorption retarder, salt, preservative, drug, Includes drug stabilizers, binders, excipients, disintegrants, lubricants, sweeteners, flavoring agents, dyes and the like and combinations thereof. Its use in therapeutic or pharmaceutical compositions is envisioned, unless conventional carriers are incompatible with the active ingredient.

The term "therapeutically effective amount" of a compound of the present invention causes a decrease or inhibition of a biological or medical response of a subject, such as enzyme or protein activity, or improves symptoms, alleviates pathology, and causes disease. Refers to the amount of a compound of the invention that slows or slows the progression or prevents the disease (eg, improves the symptoms associated with NPSLE; alleviates the pathology of NPSLE, slows or slows the progression of NPSLE).

As used herein, the term "subject" refers to an animal. Typically, the animal is a mammal. Subjects also refer to, for example, primates (eg, humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds and the like. In certain embodiments, the subject is a primatology. In yet another embodiment, the subject is a human. In certain embodiments, the human subject is also referred to as a patient.

As used herein, the terms "inhibit," "inhibit," or "inhibit" refer to a given condition, symptom, or disorder, or alleviation or suppression of a disease, or biological activity. Refers to a significant reduction in the baseline activity of the process.

As used herein, the term "treating," "treating," or "treating" any disease or disorder, in one embodiment, ameliorating the disease or disorder (ie, disease or disorder. (To slow down, stop, or alleviate the onset of at least one of its clinical symptoms). In another embodiment, "treating," "treating," or "treating" refers to reducing or ameliorating at least one physical parameter, including one that is not recognizable by the patient. In yet another embodiment, "treating," "treating," or "treating" is physically (eg, stabilizing recognizable symptoms), physiologically (eg, stabilizing physical parameters). (), Or both, refers to the regulation of disease or disorder. In one embodiment, treating NPSLE refers to treating various neurological and / or behavioral clinical manifestations of a patient suffering from systemic lupus erythematosus (SLE). These neurological and / or behavioral clinical symptoms include 19 symptoms according to the definition of the American College of Rheumatology (Artritis Rheum 1999, 42 (4): 599-608). : Aseptic meningitis, cerebrovascular disorder, demyelinating syndrome, headache, movement disorder, myelopathy, paroxysmal disorder, acute confusion, anxiety disorder, cognitive dysfunction, neuropathy, mood disorder and psychotic disorder and peripheral Sexual Syndrome Gillan Valley Syndrome, Autonomous Neuropathy, Mononeuropathy, Severe Myasthenia, Cerebral Neuropathy, Neuropathy Disorders and Multiple Neuropathies.

As used herein, the term "preventing" refers to delaying the onset or onset or progression of a disease or disorder (ie, the initiation of neurological and / or behavioral clinical manifestations as defined above. Or to delay the onset or progression).

As used herein, such a subject requires such a procedure if the subject can benefit from the procedure biologically, medically or in quality of life. ".

As used herein, the terms "a", "an", "the" and similar terms used in the context of the present invention (especially in the claims) are used herein. Unless otherwise indicated in, or unless there is a clear contradiction in the context, it should be construed to include both singular and plural.

All methods described herein can be performed in any suitable order, unless otherwise indicated herein or where there is no apparent contradiction in the context. The use of any example, or exemplary language (eg, "etc.") set forth herein, is only intended to further articulate the invention and is within the scope of the invention otherwise claimed. It does not impose any restrictions.

Any asymmetric atom (eg, carbon, etc.) of the compounds of the invention exists in a racemic or enantiomer-rich, eg, (R)-, (S)-or (R, S) -configuration. obtain. In certain embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, and at least 70% enantiomeric excess in the (R)-or (S) -configuration. , At least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess. Substituents in atoms with unsaturated bonds can be present in cis- (Z)-or trans- (E) -forms where possible.

Thus, as used herein, the use of compounds of the invention refers to possible isomers such as rotational isomers, atropisomers, racemates or mixtures thereof, such as substantially pure geometry. It can be in the form of one of (cis or trans) isomers, diastereomers, optical isomers (antipode), racemates or mixtures thereof. For clarity, positional isomers are not included by the above.

Any of the resulting mixtures of isomers, pure or substantially pure geometric or optical isomers, diastereomers, based on physicochemical differences in composition, eg, by chromatography and / or fractional crystallization. Can be separated into racemates.

Separation of the diastereomeric salt obtained by, for example, using an optically active acid or a base, and optically active acidity or basicity of either the final product or the racemate obtained as an intermediate. By liberation of the compound, it can be split into optical racemates. Thus, in particular, the basic moiety is combined with, for example, optically active acids such as tartaric acid, dibenzoyl tartaric acid, diacetyl tartaric acid, di-O, O'-p-toroil tartaric acid, mandelic acid, malic acid or camphor-10-sulfonic acid. The compounds of the present invention may be used to divide the compounds of the present invention into their optical antipode by the fractionated crystals of the salt formed. Racemic products can also be partitioned by chiral chromatography, such as high performance liquid chromatography (HPLC) or supercritical fluid chromatography (SFC), using a chiral adsorbent.

Administration and Pharmaceutical Composition In the therapeutic use of the compounds of the invention, such compounds are administered alone or as part of the pharmaceutical composition in therapeutically effective amounts. Accordingly, the present invention provides a pharmaceutical composition comprising a compound of the invention, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents, or excipients. For the purposes of the present invention, solvates and hydrates are generally considered compositions unless otherwise indicated. Preferably, the pharmaceutically acceptable carrier is sterilized.

The pharmaceutical composition of the present invention can be a unit dose of about 1 to 1000 mg of active ingredient for a subject of about 50 to 70 kg. The therapeutically effective dose of a compound, pharmaceutical composition, or combination thereof depends on the species, weight, age and individual pathology of the subject, the disorder or disease being treated, or its severity. Physicians, clinicians or veterinarians with conventional skills may readily determine the effective amount of each active ingredient required to prevent and treat the disorder or disease, or to control the progression of the disorder or disease. it can.

The above dosage characteristics can be advantageously demonstrated in in vitro or in vivo studies using mammals such as mice, rats, dogs, monkeys or excised organs, tissues and specimens thereof. The compounds of the present invention can be in vitro, in the form of a solution, eg, an aqueous solution, and in vivo, eg, as a suspension or in an aqueous solution, enteral, parenterally, and advantageously intravenously. Can be applied. In vitro doses can range from approximately ^10-3 molars to ^{10-9 molars} . The therapeutically effective amount in vivo can range from about 0.1 to 500 mg / kg, depending on the route of administration.

The pharmaceutical composition of the present invention uses a method comprising mixing the compound of the present invention, or a pharmaceutically acceptable salt thereof, with one or more pharmaceutically acceptable carriers, diluents or excipients. Can be prepared. By way of example, the pharmaceutical compositions of the invention used compounds of the invention in free or pharmaceutically acceptable salt form, along with at least one pharmaceutically acceptable carrier, diluent or excipient. Manufactured by mixing, granulation and / or coating methods.

The present invention further provides anhydrous pharmaceutical compositions and dosage forms comprising the compounds of the invention as active ingredients, as water can accelerate the degradation of certain compounds.

The anhydrous pharmaceutical composition and dosage form of the present invention can be prepared using anhydrous or low water content components and low water or low humidity conditions. Anhydrous pharmaceutical compositions can be prepared and stored so that their anhydrousity is maintained. Therefore, anhydrous compositions are packaged with materials known to prevent exposure to water so that they can be included in suitable formulation kits. Examples of suitable packaging include, but are not limited to, sealed foils, plastics, unit volume containers (eg, vials), blister packs, and strip packaging.

The present invention further provides pharmaceutical compositions and dosage forms comprising one or more agents that reduce the rate of degradation of the compounds of the invention as active ingredients. Such agents, referred to herein as "stabilizers," include, but are not limited to, antioxidants such as ascorbic acid, pH buffers, salt buffers and the like.

The pharmaceutical composition is a specific route of administration such as oral administration, rectal administration, transdermal administration, parenteral administration, intravenous administration, intramuscular administration, transpulmonary administration, inhalation administration, intranasal administration, intraocular administration and topical administration. Can be formulated for.

Oral Dosage Form The pharmaceutical composition of the present invention can be orally administered as a separate dosage form, such as, but not limited to, capsules, gelatin capsules, caplets, tablets, chewable tablets, Lozenges, dispersible powders, granules, syrups, flavored syrups, solutions or suspensions in aqueous or non-aqueous liquids, edible foams or whips, and oil-in-water emulsions or water-in-oil emulsions. A turbid liquid can be mentioned.

Therefore, for oral administration, the pharmaceutical composition of the present invention, which comprises an effective amount of the compound of the present invention, is in solid form (capsules, gelatin capsules, hard or soft capsules, tablets, chewable tablets, but not limited to). , Lozenges, capsules, pills, granules or dispersible powders), or liquid forms (including, but not limited to, solutions, aqueous or oily suspensions, syrups, elixirs, foams, whips or emulsions) Can consist of. Pharmaceutical compositions can be subjected to conventional pharmaceutical processes such as sterilization and / or conventional inert diluents, lubricants, or buffers, as well as preservatives, stabilizers, wetting agents, emulsifiers and buffers, etc. May contain an adjunct.

Compositions for oral use are prepared according to any method known in the art for the production of pharmaceutical compositions, such compositions providing a pharmaceutical-looking and palatable formulation. It may contain one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents and preservatives.

Typically, the pharmaceutical composition is
a) Diluents such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine;
b) Lubricants such as silica, talcum, stearic acid, magnesium or calcium salts thereof and / or polyethylene glycol;
c) Binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidone; as required.
d) Containing active ingredients with one or more of disintegrants such as starch, agar, alginic acid or sodium salts thereof, or effervescent mixtures; and e) absorbents, colorants, flavors and sweeteners. It is a tablet or gelatin capsule.

The tablet may contain the active ingredient in a mixture with a non-toxic, pharmaceutically acceptable excipient suitable for the manufacture of the tablet. These excipients are, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulators and disintegrants such as corn starch or alginic acid; binders such as starch. , Gelatin or acacia; and lubricants such as magnesium stearate, stearic acid or talc. The tablets may be film coated or enteric coated according to methods known in the art. Tablets are uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a lasting effect over a longer period of time. For example, a time delay material such as glyceryl monostearate or glyceryl distearate can be used. Formulations for oral use are either as hard gelatin capsules (where the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin) or as soft gelatin capsules (where here). The active ingredient may be shown as a water or oil medium (mixed with, for example, peanut oil, liquid gelatin or olive oil).

Parenteral Dosage Form In certain embodiments, the pharmaceutical compositions of the invention are administered parenterally by a variety of routes, including but not limited to, subcutaneously, intravenously including bolus injection, intramuscularly, and intraarterial. obtain.

Certain injectable compositions are aqueous isotonic solutions or suspensions, and suppositories are advantageously prepared from fat emulsions or suspensions. The composition can be sterilized and / or contains auxiliary agents such as preservatives, stabilizers, wetting or emulsifiers, solution promoters, salts and / or buffers for adjusting osmotic pressure. Can be. In addition, they may also contain other therapeutically useful substances. The composition is prepared according to each of the conventional mixing, granulation or coating methods and typically contains about 0.1-75%, or about 1-50%, active ingredient.

Topical Dosage Form In certain embodiments, the pharmaceutical composition of the invention is a topical application of a pharmaceutical composition containing a compound of the invention in the form of a lotion, gel, ointment, solution, emulsion, suspension or cream. Can be administered by.

For example, compositions suitable for topical application to the skin and eyes include aqueous solutions, suspensions, ointments, creams, gels or spray formulations for delivery, such as with aerosols. Such a topical delivery system would be particularly suitable for skin application, for example for the treatment of skin cancer, for prophylactic use in sunscreen creams, lotions, sprays and the like. Therefore, they are particularly suitable for topical use, including cosmetics and formulations well known in the art. It may contain solubilizers, stabilizers, tonic accelerators, buffers and preservatives.

As used herein, topical application may also be associated with inhalation or intranasal application. They involve the use of a dry powder form from a dry powder inhaler (either alone or as a mixture, eg, a dry blend with lactose, or, for example, as a mixed component particle with a phospholipid) or a suitable propellant. It can usually be delivered in the form of an aerosol spray from a pressurized vessel, pump, spray, injector or atomizer, with or without it.

Rectal Administration In certain embodiments, the pharmaceutical compositions of the present invention may be administered rectally in the form of suppositories, enema, ointments, creams, rectal foams or rectal gels. In certain embodiments, such suppositories are prepared from fat emulsions or suspensions, cocoa butter or other glycerides.

Depot Administration In certain embodiments, the pharmaceutical composition of the present invention of the present invention can be formulated as a depot formulation. Such formulations are administered by injection (eg, subcutaneously or intramuscularly) or by intramuscular injection. In certain embodiments, such formulations include polymers or hydrophobic materials (eg, as emulsions in acceptable oils) or ion exchange resins, or as sparingly soluble derivatives, eg, sparingly soluble salts.

In one embodiment, the compound of formula (I) or (II) according to any one of embodiments 1-12 is preferably administered by the oral route.

Combination Therapies The compounds and pharmaceutical compositions of the invention provided herein are selected from one or more additional therapeutic agents, such as anti-CD40 antibodies, such as low molecular weight antimalarial agents or biopharmacy. Can be administered in combination with.

The combination of the present invention can be a unit dose of about 1 to 1000 mg of active ingredient for a subject of about 50 to 70 kg. The therapeutically effective dose of the combination depends on the species, weight, age and individual condition of the subject, the disorder or disease being treated or its severity. Physicians, clinicians or veterinarians with conventional skills may readily determine the effective amount of each active ingredient required to prevent and treat the disorder or disease, or to control the progression of the disorder or disease. it can.

The above dosage characteristics can be advantageously demonstrated in in vitro or in vivo studies using mammals such as mice, rats, dogs, monkeys or excised organs, tissues and specimens thereof. The compounds of the present invention can be in vitro, in the form of a solution, eg, an aqueous solution, and in vivo, eg, as a suspension or in an aqueous solution, enteral, parenterally, and advantageously intravenously. Can be applied. In vitro doses can range from approximately ^10-3 molars to ^{10-9 molars} . The therapeutically effective amount in vivo can range from about 0.1 to 500 mg / kg, depending on the route of administration.

The compounds of the present invention may be administered simultaneously with, or before or after, one or more other therapeutic agents. The compounds of the invention can be administered separately, by the same or different routes of administration, or together in the same pharmaceutical composition as other agents. Therapeutic agents are, for example, chemical compounds, peptides, antibodies, antibody fragments or nucleic acids, which are therapeutically active or, when administered to a patient in combination with a compound of the invention, enhance therapeutic activity.

Further therapeutic agents used in combination with the compounds of the invention include, but are not limited to, anti-inflammatory drugs, immunomodulators, immunosuppressants, cytokines, non-steroidal anti-inflammatory drugs (NSAIDs), anti-malaria compounds, anti-malaris. Examples include rheumatic compounds, B cell activating factor (BAFF) inhibitors, B lymphocyte stimulating factor (BLyS) inhibitors, and steroid hormones, neurological agents and anti-coagulants.

Nonsteroidal anti-inflammatory drugs (NSAIDs) used in combination with the compounds of the present invention include, but are not limited to, salicylic acid, acetylsalicylic acid, methyl salicylate, diflunisal, salsalate, orsalazine, sulfasalazine, ketoprofen, indomethacin, slindac. , Etdrac, mephenamic acid, meclophenamate sodium, tolmethin, ketrolac, diclofenac, ibuprofen, naproxen, naproxen sodium, phenoprofen, ketoprofen, flurubiprofen, oxaprozine, piroxicam, meroxycam, ampyroxicam, droxycam, piroxicam, tenoxycam , Nabumetone, phenylbutazone, oxyphenbutazone, antipyrine, aminopyrin, apazone and nimeslide.

Anti-rheumatic compounds used in combination with the compounds of the present invention include, but are not limited to, methotrexate.

Antimalarial compounds used in combination with the compounds of the present invention include, but are not limited to, chloroquine and hydroxychloroquine.

The inhibitor of B cell activating factor (BAFF), also known as an inhibitor of B lymphocyte stimulating factor (BLyS), used in combination with the compounds of the present invention is, but is not limited to, belimumab ( Benlysta®, Brishibimod and BR3-Fc.

Immunosuppressants used in combination with the compounds of the present invention include, but are not limited to, mycophenolate mofetil (MMF), mycophenolic acid, cyclophosphamide, azathioprine and lacinimod (5-chloro-N-ethyl-). 4-Hydroxy-1-methyl-2-oxo-N-phenyl-1,2-dihydroquinolin-3-carboxamide) can be mentioned.

Steroid hormones used in combination with the compounds of the present invention include, but are not limited to, dehydroepiandrosterone (DHEA).

Combinations with neuropharmaceuticals, including, but not limited to, antidepressants, antiepileptic drugs, antipsychotics, may be envisioned, depending on the particular neuropsychiatric symptoms of NPSL. Since thrombus is suspected as one of the causes of NPSLE, combination with antiplatelet drug therapy or anticoagulant can be envisioned.

Experimental Part Preparation of Compounds for Use in the Invention:
Synthesis of compounds of formula (I) and formula (II) is described in US Pat. No. 9,126,999 and PCT Application International Publication No. 2018/047081 pamphlet, respectively. Therefore, both US Pat. No. 9,126,999 and International Publication No. 2018/047081 pamphlet are incorporated by reference.

Conventional technology and importance:
Anti-type 1 interferon receptor (IFNAR) antibodies have recently demonstrated efficacy in Ph2b studies in lupus patients (Arthritis and Rheumatology, 69,376-386,2017, published by R. Furie et al.). Inhibition of IFNAR also reduced the autoimmune phenotype in the Lupus 564Igi mouse model. In this model, follicular dendritic cells (FDCs) were identified as the primary source of IFNα: autoantibodies and nucleolar immune complexes yielding IFNα from FDCs by the endosome receptor TLR7 (A. Das et al.). Published by Immunity 46, 106-119, 2017). In a related study in the 564Igi model, anti-IFNAR treatment had protective effects against CNS lesions (reactive microglia, synaptic loss, and neurotransmitter microglial phagocytosis) and behavioral phenotypes (published by Bialas et al. Nature 546, pp 539, 2017).

concept:
Based on the above, we have found that TLR7 antagonists, such as compounds of the invention, such as compounds of formula (I) or in particular compounds of formula (Ia) or formula (Ib), or compounds of formula (II), in particular. Compounds of formula (IIa) young (IIb) young (IIc) block IFNα production, restore B cell tolerance and arrest the progression of peripheral autoimmunity in the 564Igi mouse lupus model, and thus CNS. It created the scientific concept of reducing lesions.

Experiments and discussions:
The effectiveness of compounds of formula (Ib) and compounds of formula (IIc) in 564Igi mice is tested using a staggered two-step approach. It is known from the literature (see Forsbach et al, J. Immunol., 2008, 180: 3729-3738) that TLR8 does not function in rodents. However, from US Pat. No. 9,126,999 B2, it is known that their underlying compounds, including compounds of formula (Ib), are double TLR7 / 8 antagonists. It is also known from WO 2018/047081 that the compounds of formulas (II), (IIa), (IIb) or (IIc) are dual TLR7 / 8 antagonists. For this reason, as avoided above, the results of the compounds of the present invention may or may not depend on the efficacy of the compound for the TLR7 and TLR8 receptors, as TLR8 does not function in rodents. is there. As a result, the effects of the compounds of the invention obtained in the following animal experiments are not necessarily linked to TLR8.

(1) Test of ability of compound of formula (Ib) or compound of formula (IIc) to reduce IFNα-dependent autoimmunity:
564Igi mice (n = 5 per group) are treated with the compound or vehicle of formula (Ib) via food for a period of 2 weeks. The compound of formula (Ib) is a potent TLR7 antagonist, i.e., IFNα secretion from human peripheral blood mononuclear cells (PBMC) after stimulation with single-strand RNA (ssRNA) combined with a liposome transfection reagent. when determined by the inhibition, having an _{IC 50} of 35 nM. This compound, for inhibiting TNFα with _{an IC 50} of 36nM in the same assay has also TLR8 antagonist component. The compound of formula (IIc) is a potent TLR7 antagonist, i.e., as determined by the huPBMC TLR7 antagonist IFNα assay disclosed in WO 2018/04081, filed September 6, 2017. It has a 4 nM IC ₅₀ . This compound, in a similar assay, for inhibiting TNFα with _{an IC 50} of 166 nm, also has TLR8 antagonist component.

Mice are bleeding regularly and blood cells are assayed for interferon-stimulated gene (ISG) expression by qPCR. CD69 expression is measured in blood as a pharmacodynamic marker after in vitro R848 stimulation to determine compound exposure in blood. On day 15, the mice are slaughtered and the spleen and lymph nodes are taken for flow cytometric analysis and immunohistochemistry. This analysis focuses on the frequency of autoantibody-specific (Id pos) B cells, germinal center B cells (GC) and plasmablasts, and the expression of IFNα. Spleen tissue is collected for ISG analysis. Antinuclear antibody titers are determined from serum by ELISA.

Experimental Results: ISG and in vitro stimulation CD69 were inhibited in 564 Igi mice treated with compound of formula (Ib) or compound of formula (IIc) as opposed to the vehicle group, which resulted in the 564 Igi model being TLR7. It is confirmed that it is a dependency.

(2) Test of ability of compound of formula (Ib) or compound of formula (IIc) to reduce NPSLE symptoms:
564Igi and NZB / WF1 mice (n = 10 per group) were subjected to a compound of formula (Ib) or a compound of formula (IIc) or a compound of formula (IIc) over a period of 6 weeks from 8 weeks of age to 14 weeks of age. Treat with vehicle. Similar to Experiment 1 above, mice are periodically bleeded and blood cells are assayed for interferon-stimulated gene (ISG) expression by qPCR. CD69 expression is measured in blood as a pharmacodynamic marker after in vitro R848 stimulation to determine compound exposure in blood. Starting at 12 weeks of age, mice are subjected to behavioral tests such as an elevated cross maze, a novel Y-shaped maze, a three-chamber social interaction assay, and a prepulse inhibition test. At the end of the experiment, mice were slaughtered to collect serum, cerebrospinal fluid (CSF), spleen, and brain tissue for the following analyzes. : The spleen is analyzed for ISG expression by qPCR and for autoantibody-specific (Idpos) B cells, germinal center B cells (GC) and plasmablasts by immunohistochemistry and flow cytometry. Antinuclear antibody titers are determined from serum by ELISA. The brain is analyzed for ISG by qPCR and microglia are tested by immunohistochemistry and flow cytometry. In addition, CD69 expression is measured in blood as a pharmacodynamic marker after in vitro R848 stimulation to determine compound exposure in blood and metabolite levels in serum, brain and CSF.

Experimental Results: In contrast to the mice in the vehicle group, 564Igi or NZB / WF1 mice treated with compounds of formula (Ib) or compounds of formula (IIc) showed reduced progression of NPSLE symptoms, ie, various. Shows reduced abnormal behavior in behavioral tests, reduced histological and molecular changes to CNS showing interferon signaling and neuropathological symptoms, and lower amounts of potentially neurotoxic metabolites. In parallel with these neuropathological findings, lower titers of antinuclear antibody and antibodies to neural components in peripheral blood and lower expression of ISG in the spleen. Overall, weaker neuropathological findings in the 564Igi or NZB / WF1 animal model of NPSLE after treatment with compounds of formula (Ib) or compounds of formula (IIc) are the etiology of TLR7, especially in NPSLE. Can show a role.

3) Neuroprotective effects provided by TLR7 / 8 antagonists in a mouse model of NPSLE Diagnosis of NPSLE defines a subgroup of SLE patients with CNS disorders. The Lupus NZBW / F1 mouse model exhibited neuropathological symptoms (Bialas et al., Lesion 2017 546 (7659): 539-543; Kier et al. J Comp Pathol 1990, 102 (2) "165-177. ), The compounds of the formula provide protection for this model, eg, systemic readout in renal lesions (Fig. 1).

Figure 1: Efficacy of compounds of formula (Ib) for systemic disease parameters in the NZBW / F1 model. NZBW / F1 mice were started at 28 weeks of age and orally (po.) At 100 mg / kg of the compound of formula (Ib) (black circles) or vehicle (white circles) once daily (q.d.). ) Treated. Proteinuria (Uristix, Bayer, Leverkusen, Germany) and body weight of NZB / W F1 mice were recorded weekly throughout the experiment, starting at 20 weeks of age. At a proteinuria score of 2 (score 0: no protein detected, 1: trace amount, 2:> 30 mg / dl, 3:> 100 mg / dl, 4:> 300 mg / dl, 5:> 2000 mg / dl) Animals were randomly divided into experimental groups with n = 15. When weight loss reached 20% or the animals had a proteinuria score of 5 for the second time in a row, individual animals were removed from the experiment and their final proteinuria score was recorded for the rest of the experiment. The study was stopped as soon as 50% of the animals in either group were removed from the experiment. A, survival, p = 0.02, log rank test, B, proteinuria, ^* , p <0.05, ^** , p <0.01, ^*** p <0.001, ^*** , p <0.0001, unpaired t-test. The points represent the mean ± SEM.

Compounds of formula (IIc) were found to reduce specific markers of brain lesions in this model. The compound of formula (IIc) blocked systemic reading (FIGS. 2a, 2b) and dose-dependently inhibited the expression of NMDAR autoantibodies (FIG. 2c), which expression of NMDAR autoantibodies was NPSLE in humans. (Omdal et al., Eur J Neurol 2005 12 (5): 392-398; Robbins et al., Artritis Rheum 1998 31 (5): 623-31), which directly disrupts neural function (Dose et al.). , PNAS 2010, 107 (43): 18569-74). In addition, compounds of formula (IIc) block circulating NF-L levels (FIG. 2d), where NF-L is a widely used clinical biomarker of inflammatory injuries to CNS (eg, Cai). et al., Neuropsychiator Dis. Treat, 2018 14: 2241-2254). The compound of formula (IIc) inhibited the blood levels of anti-NMDAR autoantibodies and NF-L, demonstrating that the compound of formula (IIc) has a neuroprotective role in NPSLE.

Materials and Methods Female 12-week-old NZB / WF1 mice (NZBNZWF1 / OlaHsd) were obtained from Envigo RMS Limited (Blackthorn, Bicester, UK). 6:00 a. m. ~ 6:00 p. m. Mice were bred (3-5 mice / individually ventilated cages, IVC) at 21 ± 2 ° C. and 55 ± 10% relative humidity in the time-controlled light period of the year and fed freely. The following opportunistic bacterial infections were reported in the Health Monitoring Report (Report Reference: 17-9212; from January 4, 2017) from Envigo: Escherichia coli, Lactobacillus (Lactobacillus) Lactobacillus spp), Staphylococcus spp (including Staphylococcus aureus), Streptococcus spp (including α-hemolytic streptococcus). At 15 weeks of age, radio frequency identification transponders (T-SL Slim Polymer Microchip, DataMars) were subcutaneously injected into the nape with a lancet under isoflurane anesthesia to uniquely identify the mice. All tests were conducted in strict compliance with Swiss law for animal protection and were approved by the Veterinary Office Basel City (Animal License No. BS-2868).

Body Weight and Proteinuria Monitoring Body weight was recorded weekly from arrival to 20 weeks of age. Starting at 20 weeks of age, body weight was recorded twice a week and proteinuria was measured weekly until the end of the experiment. Drug treatment was initiated when the mice were 23 weeks old.

For weekly proteinuria measurements, urine was collected in a glass capillary by rubbing the abdomen and immediately transferred to an ice-cold 96-well plate. Quantitative protein assay (Bio-Rad, Hercules, CA #) using bovine serum albumin (BSA) (Sigma-Aldrich, St. Louis, MO, A-8806) as a standard for protein levels in fresh urine. 5000006) was used for measurement. Mice were assigned to different treatment groups when most reached proteinuria levels of about 0.5 mg / ml.

Individual mice were removed from the experiment when weight loss reached 20% compared to 10 days ago or when animals had proteinuria levels of 10 mg / ml or higher twice in a row (every 24-48 hours). Fifty percent of the animals in the control group (also called the "vehicle" group, fed a drug-free diet) stopped the study as soon as they developed clinically relevant proteinuria (≧ 5 mg / ml).

Preparation of compound-filled food (compound of formula (IIc)) The compound of formula (IIc) was mixed into the diet of mice as follows: The compound of formula (IIc) was added to the dry powdered food. (Maus / Ratte Hartung "GLP" NAFAG3890 and NAFAG3302, Providemi Kliba, Kaiseraugst, Switzerland over the last 2.5 weeks of the experiment). Water (50% w / w) was added to obtain a homogeneous dough, which was extruded into pellets with a length of 1.5-2.0 cm and cut. The pellet was dehydrated at 37 ° C. for 24 hours in a dehydrator (Excalibur EXC10EL, Sacramento, CA). Drug-free control pellets were prepared in parallel.

A pilot PK / PD test in female NZB / WF1 mice with dietary compounds suggested 0.03% in food as the minimum fully effective dose based on in vitro blood PD readings. Based on this, 0.01% and 0.1% compounds of formula (IIc), ie 0.1 or 1 g / kg food, were selected for this test.

Mice were fed fresh food every two weeks and daily food consumption per mouse was calculated by dividing food intake by the number of days and the number of mice per cage. Food consumption was defined as the food given minus the food left in the cage.

Treatment group At 23 weeks of age, mice were assigned to one of four treatment groups:
(1) 0.01% of the compound of formula (IIc) in food (n = 14), proteinuria at the start 0.5 mg / ml,
(2) 0.1% of the compound of formula (IIc) in food (n = 14), proteinuria at the start 0.5 mg / ml,
(3) Control food (n = 14), no compound added, proteinuria at start 0.5 mg / ml (ie, "vehicle" group)
Treatment of all mice was started on the same day and maintained until the end of the experiment, 41 weeks of age. Blood was collected for drug exposure (PK), PD and ANA measurements at the indicated time points during the experiment.

1. 1. Measurement of Autoantibodies in Serum NZB / WF1 mice were bleeding from the tail vein (living) or by cardiac puncture (terminal stage) and placed in a microtube Z-gel (Sarsteadt, Nuembrecht, Germany). Serum was isolated by centrifugation (10000 g, 10 minutes, room temperature) and kept at -80 ° C.

A 100 μg / ml salmon sperm DNA diluted on a Nunc Maxisorp plate (Huberlab, Aesch, Switzerland) for the detection of anti-N-methyl-D-aspartate receptor (NMDAR) subunit 2A (NR2A) antibody. Thermo Fisher Scientific, Waltherm, MA), 10 μg / ml calf thoracic histone (Sigma-Aldrich, St. Louis, MI), 1 unit / well of ribosome P (Immunovision, Springdale, AK), 1 unit / well of Sim. Antigen (Immunovision) or 20 μg / ml DWEYS peptide (Ac-Asp-Trp-Glu-Tyr-Ser-Val-Trp-Leu-Ser-Asn-AEEAc-NHz trifluoroacetate, Bachem, Bubendorf, Switzerland) It was coated in PBS at 4 ° C. overnight. Plates were washed in PBS / 0.05% Tween-20 (Bio-Rad) and incubated with PBS / 1% v / v BSA (Merck, Darmstadt, Germany) for 1 hour at room temperature and PBS / Washed in 0.05% Tween-20. Serum dilutions of 1: 100 or 1: 300 in PBS / 1% BSA (100 μl) were added at room temperature for 2 hours and the plates were washed in PBS / 0.05% Tween-20 and HRP conjugated. Isotype-specific detection antibodies were added at room temperature for 2 hours (1/5000 for polyclonal goat anti-mouse IgG1, polyclonal goat anti-mouse IgG2a, polyclonal goat anti-mouse IgG3 and polyclonal goat anti-mouse IgM, all of which were Sourcen Biotechnology ( Birmingham, AL); 1/10000 for polyclonal goat anti-mouse IgG, Sigma Aldrich). The plates were washed and 100 μl / well of TMB substrate (BD Biosciences) was added. The reaction was stopped by adding 100 μl / well of 1N HCl and the OD was measured at λ = 450 nm (Spectramax M5, Molecular Devices, Sunnyvale, CA). Data were expressed as OD (450 nm) / (x-background).

An SS-A precoated plate (Elisa kit from Alpha diagnostic (San Antonio, TX)) was incubated with 100 μl of calibrator or 1/100 diluted serum. After 1 hour at room temperature, the plates were washed with wash buffer and the 1/100 goat anti-mouse Ig HRP provided in the kit was added at room temperature for 30 minutes. The plates were washed and 100 μl / well of TMB substrate was added. The reaction was stopped by adding 100 μl / well stop solution and OD was measured at λ = 450 nm (Spectramax M5).

The autoantibody level percent was calculated by expressing each autoantibody level for animals treated with each compound as a percentage, and 100% was defined as the average level of the corresponding autoantibodies in the vehicle-treated group. Percentages for anti-Smith, anti-ribosome P, and anti-NMDAR (IgG2a, IgG2, IgG) were averaged for composite scores.

2. 2. Measurement of Neurofilament Light Chain in Serum Neurofilament light chain (NF-L) in serum was measured using sandwich ELISA. 50 μl / in a 0.05 M carbonate-bicarbonate buffer (pH 9.6, Sigma Aldrich) overnight at 4 ° C. with a MultiArray 96 plate (MesoScale Diagnostics, Rockville, MD) stirred at 550 rpm. Wells were coated with 1.25 μg / ml anti-NF-L mouse monoclonal antibody (47: 3 (UD1), UmanDiagnostics, Umea, Swedish). The next day, the plates were blocked with 100 μl / well TBS / 3% BSA (both from Sigma-Aldrich) for 1 hour at room temperature with stirring (800 rpm). In the meantime, NF-L standard purified from bovine serum (Uman Diagnostics) is subjected to assay diluent (Tris-buffered saline TBS / 0.1) containing 300 μg / ml HeteroBlock (Omega Biologicals, Bozeman, MT). Prepared in% Tween-20 / 1% BSA) to obtain a continuous double 10 point standard curve in the range 4000 pg / ml to 7.8125 pg / ml. Plates are washed with TBS / 0.1% Tween-20 and 25 μl / well assay diluent containing 300 μg / ml HeteroBlock for standard wells or 600 μg / ml HeteroBlock for sample wells is added. It was. Standard or undiluted samples (25 μl / well) were added to the plates and incubated for 2 hours at room temperature with stirring (800 rpm). Plates were washed with TBS / 0.1% Tween-20 and 25 μl / well of 0.1875 μg / ml biotinylated anti-NFL mouse monoclonal antibody (2: 1 (UD3), Uman Diagnostics) in assay diluent. Was added. Plates are incubated for 1 hour at room temperature with stirring (800 rpm), washed with TBS / 0.1% Tween 20 and 25 μl / well of sulfo-TAG-labeled streptavidin (0.25 μg / ml,) in assay diluent. Meso Scale Diagnostics) was added. The plates were incubated for 1 hour at room temperature with stirring (800 rpm), washed with TBS / 0.1% Tween-20, and 150 μl / well of Read Buffer T (Meso Scale Diagnostics) was added. Electrochemiluminescence was quantified using the Meso Vector S600 Reader (Meso Scale Diagnostics).

Other listed embodiments Embodiment 1a. A method of treating and / or preventing NPSLE that is effective for patients in need of it, formula (II):

(During the ceremony
L is -CH _2- or -CH ₂ CH _2- ;
R ¹ is -NHC (= O) R ⁶ , -NHC (= O) (CH ₂ ) _n R ⁶ , -NHC (= O) (CH ₂ ) _m NHR ⁵ , -NHC (= O) (CH _{2) ^{) m n (R 5) 2}} , -NHC (= O) (CHR 7) m NHR 5, -NHC (= O) (CH 2) m NH 2, -NHC (= O) (CH 2) n OR 7 , -NHC (= O) OR ⁷ , -NHC (= O) (CHR ⁷ ) _n R ⁶ , -NHC (= O) (CHR ⁷ ) _n N (R ⁸ ) ₂ , -NHC (= O) (CHR) ⁷ ) _n NHR ⁸ , -NR ⁷ C (= O) OR ¹¹ , -NHC (= O) (CH ₂ ) _n N (CD ₃ ) ₂ , -NR ⁷ C (= O) R ⁵ , -NR ⁷ C (= O) (CH ₂ ) _n R ⁵ , -NR ⁷ C (= O) OR ⁵ , -NHS (= O) ₂ R ⁵ , -NHC (= O) (CH ₂ ) _n NR ⁷ C (= O) ) R ⁵ or -NHC (= O) (CH ₂ ) _n NR ⁷ S (= O) ₂ R ⁵ ;
R ² is, _H, be a C 1 -C ₆ alkyl or _C 1 -C ₆ haloalkyl;
R ³ is H, C _{1 to} C ₆ alkyl or -CD ₃ ;
R ⁴ is H, NH ₂ , C _{1 to} C ₆ alkyl or halo;
Each R ⁵ is independently selected from C _1- C ₆ alkyl, -CD ₃ and- (CH ₂ ) _n OR ⁷ ;
R ⁶ is _{independent of C 3 to} C ₆ cycloalkyl, or N, NH, N (C _{1 to} C ₆ alkyl) and O (unsubstituted or substituted with one or two R ⁹ groups). It is a 4- to 6-membered heterocycloalkyl having 1-2 ring members selected;
Each R ⁷ is independently selected from H and C _1- C _{6 alkyl;}
Each R ⁸ is independently selected from C _{1 to} C ₆ haloalkyl,-(C (R ⁷ ) ₂ ) _n OR ⁷ _{, and C 1 to} C ₆ alkyl substituted with 1 to 3 -OH;
Each ^{R 9} is _{independently} selected C 1 -C ₆ alkyl, hydroxyl, halo, and _C 1 -C ₆ alkyl substituted with 1 to 3 -OH;
n is 1, 2, 3, 4, 5 or 6;
m is 1, 2, 3, 4, 5 or 6); or a method comprising administering a pharmaceutically acceptable salt thereof.

Embodiment 2a. The compound is of formula (IIa):

(During the ceremony
R ² is, _H, be a C 1 -C ₆ alkyl or _C 1 -C ₆ haloalkyl;
R ³ is H, C _{1 to} C ₆ alkyl or -CD ₃ ;
R ⁴ is H, NH ₂ , C _{1 to} C ₆ alkyl or halo;
R ⁶ is _{independent of C 3 to} C ₆ cycloalkyl, or N, NH, N (C _{1 to} C ₆ alkyl) and O (unsubstituted or substituted with one or two R ⁹ groups). It is a 4- to 6-membered heterocycloalkyl having 1-2 ring members selected;
Each R ⁹ is independently selected from C _{1 to} C ₆ alkyl, hydroxyl, halo, and C _{1 to} C ₆ alkyl substituted with 1 to 3 -OH); or pharmaceutically thereof. The method of embodiment 1a, which is an acceptable salt.

Embodiment 3a. The compound is of formula (IIa)
(During the ceremony
R ² is C _{1 to} C ₄ alkyl;
R ³ is a C _{1 to} C ₄ alkyl;
R ⁴ is _located at C 1 -C ₄ alkyl or halo;
R ⁶ is a compound of N, NH, N (C _{1 to} C ₆ alkyl) and a 4- to 6-membered heterocycloalkyl having 1-2 ring members selected independently of O; or a pharmacy thereof. The method according to embodiment 2a, which is a generally acceptable salt.

Embodiment 4a. The compound is of formula (IIb):

(During the ceremony
R ² is C _{1 to} C ₄ alkyl;
R ³ is a C _{1 to} C ₄ alkyl;
The method according to, or a pharmaceutically acceptable salt thereof, any one of the embodiments 1a to 3a; compound of R ⁴ is _C 1 -C ₄ alkyl).

Embodiment 5a. The compound is of formula (IIc):

The method according to any one of embodiments 1a-4a, which is a compound of; or a pharmaceutically acceptable salt thereof.

Embodiment 6a. A method of treating and / or preventing NPSLE that is effective for patients in need of it: Formula (I):

(During the ceremony
R _{1 is a C 1 to} C ₄ alkyl that is optionally substituted at least once with hydrogen or halogen;
R ₂ and R ₃ are independent of each other, hydrogen or C _{1 to} C ₆ alkoxy;
R ₄ is hydrogen or C _{1 to} C ₆ alkyl;
A method comprising administering a compound (where R ₅ is hydrogen or C _{1 to} C ₆ alkyl); or a pharmaceutically acceptable salt thereof.

Embodiment 7a. The compound is of formula (I)
(During the ceremony
R ₁ is trifluoromethyl or difluoromethyl;
R ₂ and R ₃ are both methoxy;
R ₄ is hydrogen;
R ₅ is hydrogen or compounds of the methyl a is); or a pharmaceutically acceptable salt thereof, The method of embodiment 6a.

8a. The compound has the formula (Ia).

The method according to embodiment 6a or 7a, which is a compound of the above or a pharmaceutically acceptable salt thereof.

Embodiment 9a. The compound has the formula (Ib).

The method according to embodiment 6a, 7a or 8a, which is a compound of the above or a pharmaceutically acceptable salt thereof.

Embodiment 10a. The method according to any one of embodiments 1a-9a, wherein NPSL refers to various neurological and / or behavioral clinical manifestations of a patient suffering from systemic lupus erythematosus (SLE).

Claims (15)

Formula (II) for use in the treatment and / or prevention of NPSLE

(During the ceremony
L is -CH _2- or -CH ₂ CH _2- ;
R ¹ is -NHC (= O) R ⁶ , -NHC (= O) (CH ₂ ) _n R ⁶ , -NHC (= O) (CH ₂ ) _m NHR ⁵ , -NHC (= O) (CH _{2) ^{) m n (R 5) 2}} , -NHC (= O) (CHR 7) m NHR 5, -NHC (= O) (CH 2) m NH 2, -NHC (= O) (CH 2) n OR 7 , -NHC (= O) OR ⁷ , -NHC (= O) (CHR ⁷ ) _n R ⁶ , -NHC (= O) (CHR ⁷ ) _n N (R ⁸ ) ₂ , -NHC (= O) (CHR) ⁷ ) _n NHR ⁸ , -NR ⁷ C (= O) OR ¹¹ , -NHC (= O) (CH ₂ ) _n N (CD ₃ ) ₂ , -NR ⁷ C (= O) R ⁵ , -NR ⁷ C (= O) (CH ₂ ) _n R ⁵ , -NR ⁷ C (= O) OR ⁵ , -NHS (= O) ₂ R ⁵ , -NHC (= O) (CH ₂ ) _n NR ⁷ C (= O) ) R ⁵ or -NHC (= O) (CH ₂ ) _n NR ⁷ S (= O) ₂ R ⁵ ;
R ² is, _H, be a C 1 -C ₆ alkyl or _C 1 -C ₆ haloalkyl;
R ³ is H, C _{1 to} C ₆ alkyl or -CD ₃ ;
R ⁴ is H, NH ₂ , C _{1 to} C ₆ alkyl or halo;
Each R ⁵ is independently selected from C _1- C ₆ alkyl, -CD ₃ and- (CH ₂ ) _n OR ⁷ ;
R ⁶ is _{independent of C 3 to} C ₆ cycloalkyl, or N, NH, N (C _{1 to} C ₆ alkyl) and O (unsubstituted or substituted with one or two R ⁹ groups). It is a 4- to 6-membered heterocycloalkyl having 1-2 ring members selected;
Each R ⁷ is independently selected from H and C _1- C _{6 alkyl;}
Each R ⁸ is independently selected from C _{1 to} C ₆ haloalkyl,-(C (R ⁷ ) ₂ ) _n OR ⁷ _{, and C 1 to} C ₆ alkyl substituted with 1 to 3 -OH;
Each ^{R 9} is _{independently} selected C 1 -C ₆ alkyl, hydroxyl, halo, and _C 1 -C ₆ alkyl substituted with 1 to 3 -OH;
n is 1, 2, 3, 4, 5 or 6;
m is 1, 2, 3, 4, 5 or 6)
Compound or a pharmaceutically acceptable salt thereof. The compound is of formula (IIa):

(During the ceremony
R ² is, _H, be a C 1 -C ₆ alkyl or _C 1 -C ₆ haloalkyl;
R ³ is H, C _{1 to} C ₆ alkyl or -CD ₃ ;
R ⁴ is H, NH ₂ , C _{1 to} C ₆ alkyl or halo;
R ⁶ is _{independent of C 3 to} C ₆ cycloalkyl, or N, NH, N (C _{1 to} C ₆ alkyl) and O (unsubstituted or substituted with one or two R ⁹ groups). It is a 4- to 6-membered heterocycloalkyl having 1-2 ring members selected;
Each R ⁹ is independently selected from C _{1 to} C ₆ alkyl, hydroxyl, halo, and C _{1 to} C ₆ alkyl substituted with 1-3 -OH), claim 1. A compound of formula (II) for use as described in; or a pharmaceutically acceptable salt thereof. R ² is C _{1 to} C ₄ alkyl;
R ³ is a C _{1 to} C ₄ alkyl;
R ⁴ is _located at C 1 -C ₄ alkyl or halo;
Claim 1 or 2, wherein R ⁶ is a 4- to 6-membered heterocycloalkyl having 1-2 ring members that are independently selected from N, NH, N (C _{1 to} C _{6 alkyl) and O.} A compound of formula (IIa) for use as described, or a pharmaceutically acceptable salt thereof. The compound is of formula (IIb):

(During the ceremony
R ² is C _{1 to} C ₄ alkyl;
R ³ is a C _{1 to} C ₄ alkyl;
Or a pharmaceutically acceptable salt thereof; R ⁴ is a compound of C ₁ -C ₄ alkyl), compounds of formula (IIa) for use according to any one of claims 1 to 3. The compound is of formula (IIc):

The compound of formula (II) for use according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof. Formula (I) for use in the treatment and / or prevention of NPSLE

(During the ceremony
R _{1 is a C 1 to} C ₄ alkyl that is optionally substituted at least once with hydrogen or halogen;
R ₂ and R ₃ are independent of each other, hydrogen or C _{1 to} C ₆ alkoxy;
R ₄ is hydrogen or C _{1 to} C ₆ alkyl;
A compound (where R ₅ is hydrogen or C _{1 to} C ₆ alkyl); or a pharmaceutically acceptable salt thereof. R ₁ is trifluoromethyl or difluoromethyl;
R ₂ and R ₃ are both methoxy;
R ₄ is hydrogen;
R ₅ is hydrogen or methyl, the compound or a pharmaceutically acceptable salt thereof of formula (I) for use according to claim 6. The compound has the formula (Ia).

The compound of formula (I) for use according to claim 6, which is a compound of the above, or a pharmaceutically acceptable salt thereof. The compound has the formula (Ib).

The compound of formula (I) for use according to claim 6, which is a compound of the above, or a pharmaceutically acceptable salt thereof. The formula for use according to any one of claims 1-5, wherein NPSL refers to various neurological and / or behavioral clinical manifestations of a patient suffering from systemic lupus erythematosus (SLE). A compound of II) or a pharmaceutically acceptable salt thereof. The formula for use according to any one of claims 6-9, wherein NPSL refers to various neurological and / or behavioral clinical manifestations of a patient suffering from systemic lupus erythematosus (SLE). The compound of I), or a pharmaceutically acceptable salt thereof. One or more pharmaceutically acceptable salts of the compound of formula (II) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 for use in the treatment and / or prevention of NPSLE. A pharmaceutical composition comprising with an acceptable carrier. One or more pharmaceutically acceptable salts of the compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 6 to 9 for use in the treatment and / or prevention of NPSLE. A pharmaceutical composition comprising with an acceptable carrier. Use of the compound represented by the formula (II) according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof in the production of a drug for treating NPSLE. Use of the compound represented by the formula (I) according to any one of claims 6 to 9 or a pharmaceutically acceptable salt thereof in the manufacture of a drug for treating NPSLE.