WO2001044221A1 - Hydroxamic acid derivatives as inhibitors of human cd23 and of the tnf release - Google Patents

Abstract

Compounds of formula (I): wherein R is methyl substituted by one to three groups selected from alkyl, aryl, alkenyl, and alkynyl; R1 is arylmethyl or heterocyclylmethyl; and R?2 and R3¿ are both alkyl; are useful in the treatment of disorders mediated by s-CD23.

Description

HYDROXAMIC ACID DERIVATIVES AS INHIBITORS OF HUMAN CD23 AND OF

THE TNF RELEASE

This invention relates to novel inhibitors of the formation of soluble human CD23 and their use in the treatment of conditions associated with excess production of soluble CD23 (s-CD23) such as autoimmune disease, inflammation and allergy. CD23 (the low affinity IgE receptor FceRII, Blast 2), is a 45 kDa type II integral protein expressed on the surface of a variety of mature cells, including B and T lymphocytes, macrophages, natural killer cells, Langerhans cells, monocytes and platelets (Delespesse et al, Adv Immunol, 49 [1991] 149-191). There is also a CD23-like molecule on eόsinophils (Grangette et al, J Immunol, H3 [1989] 3580-3588). CD23 has been implicated in the regulation of the immune response (Delespesse et al, Immunol Rev, 125 [1992] 77-97). Human CD23 exists as two differentially regulated isoforms, a and b, which differ only in the amino acids at the intracellular N-terminus (Yokota et al, Cell, 55 [1988] 611-618). In man the constitutive a isoform is found only on B-lymphocytes, whereas type b, inducible by IL- 4, is found on all cells capable of expressing CD23.

Intact, cell bound CD23 (i-CD23) is known to undergo cleavage from the cell surface leading to the formation of a number of well-defined soluble fragments (s-CD23), which are produced as a result of a complex sequence of proteolytic events, the mechanism of which is still poorly understood (Bourget et al J Biol Chem, 269 [1994] 6927-6930). Although not yet proven, it is postulated that the major soluble fragments (Mr 37, 33, 29 and 25 kDa) of these proteolytic events, all of which retain the C-terminal lectin domain common to i-CD23, occur sequentially via initial formation of the 37 kDa fragment (Letellier et al, JExp Med, \T2 [1990] 693-700). An alternative intracellular cleavage pathway leads to a stable 16 kDa fragment differing in the C-terminal domain from i-CD23 (Grenier-B rosette et al, EurJ Immunol, 22 [1992] 1573-1577).

Several activities have been ascribed to membrane bound i-CD23 in humans, all of which have been shown to play a role in IgE regulation. Particular activities include: a) antigen presentation, b) IgE mediated eosinophil cytotoxicity, c) B cell homing to germinal centres of lymph nodes and spleen, and d) downregulation of IgE synthesis (Delespesse et al, Adv Immunol, 49, [1991] 149-191). The three higher molecular weight soluble CD23 fragments (Mr 37, 33 and 29 kDa) have multifunctional cytokine properties which appear to play a major role in IgE production. Thus, the excessive formation of s- CD23 has been implicated in the overproduction of IgE, the hallmark of allergic diseases such as extrinsic asthma, rhinitis, allergic conjunctivitis, eczema, atopic dermatitis and anaphylaxis (Sutton and Gould, Nature, 366, [1993] 421-428). Other biological activities attributed to S-CD23 include the stimulation of B cell growth and the induction of the release of mediators from monocytes. Thus, elevated levels of S-CD23 have been observed in the serum of patients having B-chronic lymphocytic leukaemia (Sarfati et al, Blood, 71 [1988] 94-98) and in the synovial fluids of patients with rheumatoid arthritis (Chomarat et al, Arthritis and Rheumatism, 36 [1993] 234-242). That there is a role for CD23 in inflammation is suggested by a number of sources. First, sCD23 has been reported to bind to extracellular receptors which when activated are involved in cell- mediated events of inflammation. Thus, sCD23 is reported to directly activate monocyte TNF, IL-1 , and IL-6 release (Armant et al, vol 180, J. Exp. Med., 1005- 1011 (1994)). CD23 has been reported to interact with the B2-integrin adhesion molecules, CD l ib and CD l ie on monocyte/macrophage (S. Lecoanet-Henchoz et al, Immunity, vol 3; 119-125 (1995)) which trigger NO^" , hydrogen peroxide and cytokine ( IL-1, IL-6, and TNF) release. Finally, IL-4 or IFN induce the expression of CD23 and its release as sCD23 by human monocytes. Ligation of the membrane bound CD23 receptor with IgE/anti-IgE immune complexes or anti CD23 mAb activates cAMP and IL-6 production and thromboxane B2 formation, demonstrating a receptor-mediated role of CD23 in inflammation.

Because of these various properties of CD23, compounds which inhibit the formation of S-CD23 should have twofold actions of a) enhancing negative feedback inhibition of IgE synthesis by maintaining levels of i-CD23 on the surface of B cells, and b) inhibiting the immunostimulatory cytokine activities of higher molecular weight soluble fragments (Mr 37, 33 and 29 kDa) of S-CD23. In addition, inhibition of CD23 cleavage should mitigate sCD23-induced monocyte activation and mediator formation, thereby reducing the inflammatory response.

International Patent Application No. WO 96/02240 (SmithKline Beecham pic) discloses that compounds which inhibit the action of matrix metalloproteases (e.g. collagenase, stromelysin and gelatinase) are effective inhibitors of the release of human soluble CD23 transfected into mammalian cell culture systems.

Unpublished International Patent Application No. PCT/GB99/01954 (SmithKline Beecham pic) discloses that certain compounds of formula (A) are effective inhibitors of the release of human soluble CD23 transfected into mammalian cell culture systems:

O R¹ O

H

HON - ^ ^ ^N ^ ^ NH(0)_nR

O R²

OR

(A) According to the present invention, there is provided a compound of formula (I):

(I) wherein:

R is methyl substituted by one to three groups selected from alkyl, aryl, alkenyl, and alkynyl;

R.1 is arylmethyl or heterocyclylmethyl; and R² and R³ are both alkyl.

Alkyl, alkenyl and alkynyl groups referred to herein include straight and branched groups containing up to six carbon atoms and are optionally substituted by one or more groups selected from the group consisting of aryl, heterocyclyl, (Cι_6)alkylfhio, (C]_6)alkoxy, aryl(C \ _ )alkoxy, aryl(Cι_6)alkylfhio, amino, mono- or di- (Cι_6)alkylamino, cycloalkyl, cycloalkenyl, carboxy and esters thereof, hydroxy, and halogen.

Cycloalkyl and cycloalkenyl groups referred to herein include groups having between three and eight ring carbon atoms and are optionally substituted as described hereinabove for alkyl, alkenyl and alkynyl groups.

When used herein, the term "aryl" means single and fused rings suitably containing from 4 to 7, preferably 5 or 6, ring atoms in each ring, which rings, may each be unsubstituted or substituted by, for example, up to three substituents. A fused ring system may include aliphatic rings and need include only one aromatic ring.

Suitable aryl groups include phenyl and naphthyl such as 1-naphthyl or 2- naphthyl.

Suitably any aryl group, including phenyl and naphthyl, may be optionally substituted by up to five, preferably up to three substituents. Suitable substituents include halogen, (C ] _6)alky I, aryl, aryl(C \ _6)alkyl, (C i _6)alkoxy, (C i _6)alkoxy(C i _6)alkyl, halo(Cι_6)alkyl, aryl(C ι_6)alkoxy, hydroxy, nitro, cyano, azido, amino, mono- and di-N- (C]_.6)alkylamino, acylamino, arylcarbonylamino, acyloxy, carboxy, carboxy salts, carboxy esters, carbamoyl, mono- and di-N-(Cι_6)alkylcarbamoyl, (Ci_.g)alkoxycarbonyl, aryloxycarbonyl, ureido, guanidino, sulphonylamino, aminosulphonyl, (C \ .g)alkylthio, (C \ _6)alkyl sulphinyl, (C \ _6)alkylsulphonyl, heterocyclyl and heterocyclyl (Cj.^alkyl. In addition, two adjacent ring carbon atoms may be linked by a (C3_5)alkylene chain, to form a carbocyclic ring.

When used herein the terms "heterocyclyl" and "heterocyclic" suitably include, unless otherwise defined, aromatic and non-aromatic, single and fused, rings suitably containing up to four heteroatoms in each ring, each of which is selected from oxygen, nitrogen and sulphur, which rings, may be unsubstituted or substituted by, for example, up to three substituents. Each heterocyclic ring suitably has from 4 to 7, preferably 5 or

6, ring atoms. A fused heterocyclic ring system may include carbocyclic rings and need include only one heterocyclic ring.

Preferably a substituent for a heterocyclyl group is selected from halogen, (C\_

₆)alkyl, aryl(Cι_₆)alkyl, (C₁.₆)alkoxy, (Cι_₆)alkoxy(Cι_₆)alkyl, halo(C₁.₆)alkyl, hydroxy, amino, mono- and di-N-(Cι_6)alkyl-amino, acylamino, carboxy salts, carboxy esters, carbamoyl, mono- and di-N-(Cι_6)alkylcarbonyl, aryloxycarbonyl, (C\_ g)alkoxycarbonyl(Cι_6)alkyl, aryl, oxy groups, ureido, guanidino, sulphonylamino, aminosulphonyl, (Cι.g)alkylthio, (C]_6)alkylsulphinyl, (Cι_6)alkylsulphonyl, heterocyclyl and heterocyclyl(C]_6)alkyl.

In a particular aspect of the invention, R is allyl, propyl, ethyl or isopropyl, and/or

R! is 1- or 2-naphthylmethyl; and/or R² is methyl or t-butyl; and/or R³ is methyl or t- butyl. In a further aspect of the invention, each of R to R³ is selected from the group consisting of the values ascribed to it in the Examples hereinbelow. Preferably, the compound of formula (I) of the invention is selected from the group consisting of the compounds described in the Examples hereinbelow.

According to a further aspect, the present invention provides the use of a compound of formula (I) for the production of a medicament for the treatment or prophylaxis of disorders such as allergy, inflammatory disorders and autoimmune disease in which the overproduction of S-CD23 is implicated.

In a further aspect the invention provides a method for the treatment or prophylaxis of disorders such as allergy, inflammatory disorders and autoimmune disease in which the overproduction of S-CD23 is implicated, which method comprises the administration of a compound of formula (I), to a human or non-human mammal in need thereof.

The invention also provides a pharmaceutical composition for the treatment or prophylaxis of disorders such as allergy, inflammatory disorders and autoimmune disease in which the oveφroduction of S-CD23 is implicated which comprises a compound of formula (I) and optionally a pharmaceutically acceptable carrier therefor.

Particular inflammatory disorders include CΝS disorders such as Alzheimer's disease, multiple sclerosis, and multi-infarct dementia, as well as the inflammation mediated sequelae of stroke and head trauma. It is to be understood that the pharmaceutically acceptable salts, solvates and other pharmaceutically acceptable derivatives of the compound of formula (I) are also included in the present invention.

Salts of compounds of formula (I) include for example acid addition salts derived from inorganic or organic acids, such as hydrochlorides, hydrobromides, hydroiodides, p- toluenesulphonates, phosphates, sulphates, acetates, trifluoroacetates, propionates, citrates, maleates, fumarates, malonates, succinates, lactates, oxalates, tartarates and benzoates.

Salts may also be formed with bases. Such salts include salts derived from inorganic or organic bases, for example alkali metal salts such as sodium or potassium salts, and organic amine salts such as moφholine, piperidine, dimethylamine or diethylamine salts.

It has suφrisingly been found that the compounds of the present invention are potent and selective inhibitors of CD23 processing and TNF release, whilst exhibiting reduced collagenase inhibitory activity in comparison with the above-mentioned compounds of the prior art. The compounds of the invention also exhibit advantageous in-vivo absoφtion properties via the oral route.

The compounds of the invention may be prepared by use of any appropriate conventional method, for example by analogy with the methods disclosed in patent publication WO 97/02239 (BBL).

Accordingly, a further aspect of the invention provides a process for preparing a compound of formula (I) as defined hereinabove, which process comprises: (a) deprotecting a compound of formula (II):

(II) wherein R to R³ are as defined hereinabove, and X is a protecting group such as benzyl or trimefhylsilyl or

(b) reacting a compound of formula (III):

(III) wherein R to R³ are as defined hereinabove, and any hydroxy group is optionally protected, with hydroxylamine or a salt thereof, or (c) converting a compound of formula (I) to a different compound of formula

(I) as defined hereinabove.

Compounds of formulae (II) and (III) are novel and form a further aspect of the invention.

Compounds of formula (II) can be prepared from compounds of formula (III) by reaction with a protected hydroxylamine. Compounds of formula (III) having one or more protected hydroxy groups can be converted by hydrolysis to a corresponding unprotected compound of formula (III).

Suitable protecting groups for a hydroxamic acid are well known in the art and include benzyl, trimethylsilyl, t-butyl and t-butyldimethylsilyl. Suitable protecting groups for a carboxylic acid are well known in the art and include t-butyl , benzyl and methyl.

Compounds of formula (III) can be prepared by reacting a compound of formula (IV) or (IVa):

(IVa) wherein R and R are as defined hereinabove and Y is a protecting group for carboxyl, with a compound of formula (V):

(V) wherein n, R² and R³ are as defined hereinabove, or an activated derivative thereof. If (IVa) is used a subsequent alkylation or acylation of the hydroxyl group may then be required.

Compounds of formula (IV) can be prepared by protecting a corresponding compound in which Y is hydrogen, which in turn can be prepared by: (a) reacting a compound of formula (VI):

(VI) wherein R is as defined hereinabove and Z is a protecting group for carboxyl, with an alkylating agent; and

(b) removing the protecting groups.

Compounds of formula (VI) wherein Z is hydrogen can be prepared by reacting a diester (such as the dimethyl or diethyl ester) of 2-hydroxy succinic acid with a compound of formula R χ' in the presence of a strong base such as lithium diisopropylamide, wherein X' is a leaving group such as bromine or iodine, and then hydrolysing the resulting compound to remove the ester groups.

The isomers, including stereoisomers, of the compounds of the present invention may be prepared as mixtures of such isomers or as individual isomers.

The individual isomers may be prepared by any appropriate method, for example individual stereoisomers may be prepared by stereospecific chemical synthesis starting from chiral substrates or by separating mixtures of diastereoisomers using known methods. In a preferred aspect, the invention provides compounds of formula (IA):

(1A) It is preferred that the compounds are isolated in substantially pure form. As stated herein an inhibitor of the formation of soluble human CD23 has useful medical properties. Preferably the active compounds are administered as pharmaceutically acceptable compositions.

The compositions are preferably adapted for oral administration. However, they may be adapted for other modes of administration, for example in the form of a spray, aerosol or other conventional method for inhalation, for treating respiratory tract disorders; or parenteral administration for patients suffering from heart failure. Other alternative modes of administration include sublingual or transdermal administration.

The compositions may be in the form of tablets, capsules, powders, granules, lozenges, suppositories, reconstitutable powders, or liquid preparations, such as oral or sterile parenteral solutions or suspensions.

In order to obtain consistency of administration it is preferred that a composition of the invention is in the form of a unit dose.

Unit dose presentation forms for oral administration may be tablets and capsules and may contain conventional excipients such as binding agents, for example syrup, acacia, gelatin, sorbitol, tragacanth, or polyvinylpyrrolidone; fillers, for example lactose, sugar, maize-starch, calcium phosphate, sorbitol or glycine; tabletting lubricants, for example magnesium stearate; disintegrants, for example starch, polyvinylpyrrolidone, sodium starch glycollate or microcrystalline cellulose; or pharmaceutically acceptable wetting agents such as sodium lauryl sulphate. The solid oral compositions may be prepared by conventional methods of blending, filling or tabletting. Repeated blending operations may be used to distribute the active agent throughout those compositions employing large quantities of fillers. Such operations are of course conventional in the art. The tablets may be coated according to methods well known in normal pharmaceutical practice, in particular with an enteric coating.

Oral liquid preparations may be in the form of, for example, emulsions, syrups, or elixirs, or may be presented as a dry product for reconstitution with water or other suitable vehicle before use. Such liquid preparations may contain conventional additives such as suspending agents, for example sorbitol, syrup, methyl cellulose, gelatin, hydroxyethylcellulose, carboxymefhylcellulose, aluminium stearate gel, hydrogenated edible fats; emulsifying agents, for example lecithin, sorbitan monooleate, or acacia; non-aqueous vehicles (which may include edible oils), for example almond oil, fractionated coconut oil, oily esters such as esters of glycerine, propylene glycol, or ethyl alcohol; preservatives, for example methyl or propyl p-hydroxybenzoate or sorbic acid; and if desired conventional flavouring or colouring agents.

For parenteral administration, fluid unit dosage forms are prepared utilising the compound and a sterile vehicle, and, depending on the concentration used, can be either suspended or dissolved in the vehicle. In preparing solutions the compound can be dissolved in water for injection and filter sterilised before filling into a suitable vial or ampoule and sealing. Advantageously, adjuvants such as a local anaesthetic, a preservative and buffering agents can be dissolved in the vehicle. To enhance the stability, the composition can be frozen after filling into the vial and the water removed under vacuum. Parenteral suspensions are prepared in substantially the same manner, except that the compound is suspended in the vehicle instead of being dissolved, and sterilisation cannot be accomplished by filtration. The compound can be sterilised by exposure to ethylene oxide before suspending in the sterile vehicle. Advantageously, a surfactant or wetting agent is included in the composition to facilitate uniform distribution of the compound. Compositions of this invention may also suitably be presented for administration to the respiratory tract as a snuff or an aerosol or solution for a nebulizer, or as a microfine powder for insufflation, alone or in combination with an inert carrier such as lactose. In such a case the particles of active compound suitably have diameters of less than 50 microns, preferably less than 10 microns for example diameters in the range of 1-50 microns, 1-10 microns or 1-5 microns. Where appropriate, small amounts of other anti-asthmatics and bronchodilators, for example sympathomimetic amines such as isoprenaline, isoetharine, salbutamol, phenylephrine and ephedrine; xanthine derivatives such as theophylline and aminophylline and corticosteroids such as prednisolone and adrenal stimulants such as ACTH may be included.

The compositions may contain from 0.1% to 99% by weight, preferably from 10-60% by weight, of the active material, depending upon the method of administration. A preferred range for inhaled administration is 10-99%, especially 60-99%, for example 90, 95 or 99%.

Microfine powder formulations may suitably be administered in an aerosol as a metered dose or by means of a suitable breath-activated device. Suitable metered dose aerosol formulations comprise conventional propellants, cosolvents, such as ethanol, surfactants such as oleyl alcohol, lubricants such as oleyl alcohol, desiccants such as calcium sulphate and density modifiers such as sodium chloride. Suitable solutions for a nebulizer are isotonic sterilised solutions, optionally buffered, at for example between pH 4-7, containing up to 20mg/ml of compound but more generally 0.1 to lOmg/ml, for use with standard nebulisation equipment.

An effective amount will depend on the relative efficacy of the compounds of the present invention, the severity of the disorder being treated and the weight of the sufferer. Suitably, a unit dose form of a composition of the invention may contain from 0.1 to lOOOmg of a compound of the invention (0.001 to lOmg via inhalation) and more usually from 1 to 500mg, for example 1 to 25 or 5 to 500mg. Such compositions may be administered from 1 to 6 times a day, more usually from 2 to 4 times a day, in a manner such that the daily dose is from lmg to lg for a 70 kg human adult and more particularly from 5 to 500mg. That is in the range of about 1.4 x 10"2 mg/kg/day to 14 mg/kg/day and more particularly in the range of about 7 x 10^~2 mg/kg/day to 7 mg/kg/day.

The following examples illustrate the invention but do not limit it in any way.

BIOLOGICAL TEST METHODS

Procedure 1: The ability of test compounds to inhibit the release of soluble CD23 was investigated by use of the following procedure.

RPMI 8866 Cell membrane CD23 cleavage activity assay:

Plasma membranes from RPMI 8866 cells, a human Epstein-Barr virus transformed B-cell line (Sarfati et al., Immunology 60 [1987] 539-547) expressing high levels of CD23 are purified using an aqueous extraction method. Cells resuspended in homogenisation buffer (20mM HEPES pH 7.4, 150 mM NaCl, 1.5 mM MgC12, 1 mM DTT) are broken by N2 cavitation in a Parr bomb and the plasma membrane fraction mixed with other membranes is recovered by centrifugation at 10,000Xg. The light pellet is resuspended in 0.2 M potassium phosphate, pH 7.2 using 2 ml per 1-3 g wet cells and the nuclear pellet is discarded. The membranes are further fractionated by partitioning between Dextran 500 (6.4% w/w) and polyethylene glycol (PEG) 5000 (6.4% w/w) (ref), at 0.25 M sucrose in a total of 16 g per 10-15 mg membrane proteins [Morre and Morre, BioTechniques 7, 946-957 (1989)]. The phases are separated by brief centrifugation at lOOOXg and the PEG (upper) phase is collected, diluted 3-5 fold with 20 mM potassium phosphate buffer pH 7.4, and centrifuged at 100,000Xg to recover membranes in that phase. The pellet is resuspended in phosphate-buffered saline and consists of 3-4 fold enriched plasma membranes as well as some other cell membranes (e.g. lysosomes, Golgi). The membranes are aliquoted and stored at -80°C. Fractionation at 6.6 % Dextran/PEG yields plasma membranes enriched 10-fold.

The fractionated membranes are incubated at 37°C for times up to 4 hrs to produce fragments of CD23 which are separated from the membrane by filtration in 0.2 micron Durapore filter plates (Millipore) after quenching the assay with 5 uM Preparation 1 from P 30994. sCD23 released from the membrane is determined using the EIA kit from The Binding Site (Birmingham, UK) or a similar one utilising MHM6 anti-CD23 mAb [Rowe et al., Int. J. Cancer, 29, 373-382 (1982)] or another anti-CD23 mAb as the capture antibody in a sandwich EIA.. The amount of soluble CD23 made by 0.5 ug membrane protein in a total volume of 50 ul phosphate-buffered saline is measured by EIA and compared to the amount made in the presence of various concentrations of inhibitors. Inhibitors are prepared in solutions of water or dimethylsulfoxide (DMSO) and the final DMSO concentration is not more than 2 %. IC50's are determined by curve fitting as the concentration where 50 % inhibition of production of sCD23 is observed relative to the difference in sCD23 between controls incubated without inhibitor.

Procedure 2: The ability of test compounds to inhibit collagenase was investigated using the following procedure.

Collagenase inhibition assay:

The potency of compounds to act as inhibitors of collagenase was determined by the method of Cawston and Barrett (Anal. Biochem. 99, 340-345, 1979), hereby incoφorated by reference, whereby a 1 mM solution of the inhibitor being tested or dilutions thereof, was incubated at 37 °C for 18 h with collagen and human recombinant collagenase, from synovial fibroblasts cloned, expressed and purified from E. Coli, (buffered with 150 mM Tris, pH 7.6, containing 15 mM calcium chloride, 0.05% Brij 35, 200 mM sodium chloride and 0.02% sodium azide). The collagen was acetylated ³H type 1 bovine collagen prepared by the method of Cawston and Muφhy (methods in Enzymology 80, 71 1 ,1981) The samples were centrifuged to sediment undigested collagen and an aliquot of the radioactive supernatant removed for assay on a scintillation counter as a measure of hydrolysis. The collagenase activity in the presence of IraM inhibitor, or dilution thereof, was compared to activity in a control devoid of inhibitor and the results reported as that concentration effecting 50% of the collagenase (IC50).

Procedure 3: The ability of test compounds to inhibit TNF release was investigated using the following procedure.

Assay for inhibition of release of TNFα from human monocytes stimulated by lipopolysaccharide (LPS) endotoxin.

Human monocytes, cultured in RPMI 1640 medium supplemented with 10 % fetal calf serum, are centrifuged at lOOOXg for 5 min and then resuspended in medium at 2 X 10 6 cells/ ml. The cell suspension is aliquoted in 24 well plates, 1 ml per well. Compounds to be tested are dissolved in neat dimethyl sulfoxide (DMSO) and added to culture with the final DMSO concentration at 0.1 %. Compounds are added to cells in triplicate wells. TNF α release is stimulated by addition of LPS to the cells at a final concentration of 200 ng/ml. Appropriate control cultures are set up in triplicates also. The plates are incubated for 18-20 hrs at 37° C, 5% CO₂, then centrifuged at 1000 Xg for 5 min. A specific ELISA for human TNFα (SmithKline Beecham) is used to measure TNF levels in the cell-free culture supernatants.

Examples

Example 1

(2S,3R)-N-((S)-4,4-Dimethyl-2-oxo-tetrahydrofuran-3-yI)-2-hydroxy-3-naphthalen- 2-ylmethyI-succinamic acid tert-butyl ester

A mixture of (2R,3S)-2-hydroxy-3-naphthalen-2-ylmethyl acid 1 -tert-butyl ester (0.62g, 1.88 mmol), HATU (0.86g, 2.26 mmol) and (S)-3-amino-4,4-dimethyl-dihydro-furan-2- one toluene-4-sulfonic acid salt (0.68g, 2.26 mmol) in DMF (9ml) was stirred at room temperature under argon for 10 minutes and then di-isopropylefhylamine (0.82 ml, 4.70 mmol) was added via syringe. The solution was stirred overnight, the DMF was evaporated and the residue was partitioned between ethyl acetate and aqueous sodium bicarbonate. The organic layer was washed with 0.5 M HCl, water and saturated brine and then dried (MgSOz).) and concentrated. Azeotroping with toluene gave 0.81 g of the desired compound (97 % yield). MS (ES +ve) M+H = 442. ^lR NMR (DMSO-d₆): 0.90 (3H, s), 1.04 (3H, s), 1.41 (9H, s), 2.93 (IH, m), 3.10 (2H, m), 3.91 (IH, m), 3.96 (IH, d, J = 8.2 Hz), 4.06 (IH, d, J = 8.2 Hz), 4.74 (IH, d, J = 9.0 Hz), 5.56 (IH, d, J = 7.1 Hz), 7.38 (IH, d, J = 8.5 Hz), 7.46 (2H, m), 7.71 (IH, s), 7.84 (3H, m), 8.37 (IH, d, J = 9.0 Hz).

Example 2

(2S,3R)-2-Allyloxy-N-((S)-4,4-dimethyl-2-oxo-tetrahydro-furan-3-yI)-3-naphthalen- 2-ylmethyl-succinamic acid tert-butyl ester

To a solution of (2S,3R)-N-((S)-4,4-Dimethyl-2-oxo-tetrahydro-furan-3-yl)-2-hydroxy-3- naphtalen-2-ylmethyl-succinamic acid tert-butyl ester (0.8 lg, 1.84 mmol) acetonitrile (14 ml) containing a few drops of DMF at room temperature under argon was added allyl bromide (1.59 mL, 18.4 mmol), followed by sodium hydride (0.096g , 2.39 mmol of 60% dispersion in oil). The reaction mixture was stirred for 1.5 hours and then quenched with a few ml of saturated ammonium chloride solution. The solvent was removed, and the residue partitioned between ethyl acetate and water. The organic layer was washed with saturated brine, dried over sodium sulphate, concentrated to a yellow oil which was chromatographed on silica gel (ethyl acetate/ hexane (1 :4) to give 0.173g of the desired compound as a yellow solid ( 20%). MS (ES +ve) M+NH4 = 504. !H NMR (DMSO-d₆): 0.86 (3H, s), 1.01 (3H, s), 1.46 (9H, s), 2.77 (IH, m), 3.05 (IH, m), 3.10 (IH, m), 3.98 (5H, m), 4.70 (IH, d, J = 8.9 Hz), 5.16 (IH, dd, J = 1.8, 10.4 Hz), 5.27 (IH, dd, J = 1.8, 17.2 Hz), 5.85 (IH, m), 7.31 (IH, dd, J = 1.6, 8.4 Hz), 7.44 (2H, m), 7.67 (IH, s), 7.80 (3H, m), 8.40 (IH, d, J = 8.9 Hz).

Example 3

(2S,3R)-N-((S)-4,4-Dimethyl-2-oxo-tetrahydro-furan-3-yI)-3-naphthaIen-2-ylmethyl- 2-propoxy-succinamic acid tert-butyl ester

A solution of (2S,3R)-2-allyloxy-N-((S)-4,4-dimethyl-2-oxo-tetrahydro-furan-3-yl)-3- naphthalen-2-ylmethyl-succinamic acid tert-butyl ester (0.17g, 0.35 mmol) in MeOH (14 ml) and Pd/BaSO (0.02g) was stirred at room temperature under hydrogen for 1 hour. The solution was then filtered through celite and the solvent evaporated to give 0.14g of a white solid (83%)

MS (ES +ve) M+Na = 506.

!H NMR (DMSO-d₆): 0.86 (3H, t, J - 7.4 Hz), 0.87 (3H, s), 1.02 (3H, s), 1.45 (9H, s),

1.49 (2H, m), 2.78 (IH, m), 3.09 (2H, m), 3.34 (2H, m), 3.84 (IH, d, J = 8.8 Hz), 3.94

(IH, d, J = 8.6 Hz), 4.02 (IH, d, J = 8.7 Hz), 4.69 (IH, d, J = 8.9 Hz), 7.31 (IH, d, J = 9.5

Hz), 7.46 (2H, m), 7.67 (IH, s), 7.81 (3H, m), 8.35 (IH, d, J = 8.7 Hz).

Example 4

(2S,3R)-N-((S)-4,4-Dimethyl-2-oxo-tetrahydro-furan-3-yl)-3-naphthalen-2-ylmethyl- 2-propoxy-succinamic acid.

A solution of (2S,3R)-N-((S)-4,4-Dimethyl-2-oxo-tetrahydro-furan-3-yl)-3-naphthalen-2- ylmethyl-2-propoxy-succinamic acid tert-butyl ester (0.14g, 0.29 mmol) of in dichloromethane/TFA (2: 1) (9 ml) was stirred at room temperature for 1.5 hours. The solvent was evaporated and the residue was azeotroped with chloroform and then toluene. The compound was triturated with hexane to give 0.12g of an off white solid (97%). MS (ES +ve) M+H = 428.

!H NMR (DMSO-d6): 0.85 (6H, m), 1.02 (3H, s),1.50 (2H, m), 2.81 (IH, m), 3.10 (2H, m), 3.27 (2H, m), 3.90 (IH, d, J = 8.6 Hz), 3.94 (IH, d, J = 9.3Hz), 3.98 (IH, d, J = 8.5 Hz), 4.70 (IH, d, J = 8.9 Hz), 7.32 (IH, dd, J = 1.7, 8.4 Hz), 7.46 (2H, m), 7.68 (IH, s), 7.80 (3H, m), 8.33 (IH, d, J = 9.3 Hz), 12.95 (IH, s).

Example 5

(2S,3R)-N'-((S)-4,4-DimethyI-2-oxo-tetrahydro-furan-3-yl)-N -hydroxy-2- naphthalen-2-ylmethyI-3-propoxy-succinamide.

A mixture of (2S,3R)-N-((S)-4,4-Dimethyl-2-oxo-tetrahydro-furan-3-yl)-3-naphthalen-2- ylmethyl-2-propoxy-succinamic acid (0.12g, 0.28 mmol) , EDC (0.1 lg, 0.56 mmol), and HO AT (0.08g, 0.56 mmol) was stirred in dry DMF at room temperature under argon for 10 minutes. Hydroxylamine hydrochloride (0.06g, 0.84 mmol) and N-methylmoφholine (0.09 ml, 0.84 mmol) were added and the reaction mixture was stirred for 4 hours. The solvent was evaporated and the residue was partitioned between ethyl acetate and water. The organic layer was washed with saturated sodium bicarbonate solution, saturated brine, dried (sodium sulphate) and concentrated to give an orange solid.

The crude compound was purified by preparative HPLC to give 48.4 mg of a white solid.(39%)

MS (ES +ve) M+H = 443, M+Na = 465

MS (ES -ve) M-H = 541. ^lH NMR (DMSO-d₆): 0.82 (3H, t, J = 7.2 Hz), 0.86 (3H, s), 1.02 (3H, s), 1.49 (2H, m), 2.97 (IH, m), 3.21 (2H, m), 3.31 (2H, m), 3.78 (IH, d, J = 9.6 Hz), 3.92 (IH, d, J = 8.4 Hz), 3.99 (IH, d, J = 8.4 Hz), 4.68 (IH, d, J = 8.8 Hz), 7.29 (IH, dd, J = 1.6, 8.8 Hz), 7.44 (2H, m), 7.67 (IH, s), 7.77 (IH, d, J = 8.4 Hz), 7.82 (2H, d, J = 6.8 Hz), 8.33 (IH, d, J = 9.2 Hz), 9.12 (lH, s), 10.98 (IH, s),

Claims

Claims

A compound of formula (I):

(I) wherein:

R is methyl substituted by one to three groups selected from alkyl, aryl, alkenyl, and alkynyl;

R is arylmefhyl or heterocyclylmethyl; and R² and R³ are both alkyl.

2. A compound according to claim 1, wherein R is allyl, propyl, ethyl or isopropyl, and/or R is 1- or 2-naphthylmethyl; and/or R² is t-butyl; and/or R³ is methyl.

3. A compound according to claim 2, wherein each of R to R is selected from the group consisting of the values ascribed to it in the Examples hereinabove.

4. A compound according to claim 2, selected from the group consisting of the compounds described in the Examples hereinabove.

5. A compound according to claim 1 or 2, which is a compound of formula (IA):

(IA)

6. The use of a compound according to any one of the preceding claims for the production of a medicament for the treatment or prophylaxis of disorders such as allergy, inflammatory disorders and autoimmune disease in which the oveφroduction of S-CD23 is implicated.

7. A method for the treatment or prophylaxis of disorders such as allergy, inflammatory disorders and autoimmune disease in which the oveφroduction of S-CD23 is implicated, which method comprises the administration of a compound according to any one of claims 1 to 5 to a human or non-human mammal in need thereof.

8. A pharmaceutical composition for the treatment or prophylaxis of disorders such as allergy, inflammatory disorders and autoimmune disease in which the oveφroduction of S-CD23 is implicated which comprises a compound according to any one of claims 1 to 5 and optionally a pharmaceutically acceptable carrier therefor.

9. A process for preparing a compound according to any one of claims 1 to 5, which process comprises:

(a) deprotecting a compound of formula (II):

(II) wherein R to R³ are as defined hereinabove, and X is a protecting group such as benzyl or trimethylsilyl or

(b) reacting a compound of formula (III):

(III) wherein R to R³ are as defined hereinabove, and any hydroxy group is optionally protected, with hydroxylamine or a salt thereof, or (c) converting a compound of formula (I) to a different compound of formula

(I) as defined hereinabove.

10. A compound of formula (II) as defined in claim 9.

1 1. A compound of formula (III) as defined in claim 9.