JP2005524630A - Inhibition of cartilage destruction via estrogen receptor - Google Patents

Abstract

The present invention relates to the pharmaceutical use of selective estrogen receptor modulators (SERMs) alone or in combination with progestins to treat or prevent diseases associated with progressive cartilage destruction. In particular, the present invention relates to the pharmaceutical use of a combination of a chroman derivative and moatindron for the treatment or prevention of osteoarthritis or rheumatoid arthritis.

Description

  The present invention relates to selective estrogen receptor modulators (SERMs) alone or in combination with other pharmaceuticals, particularly progestins, to treat or prevent a wide range of diseases, but particularly those associated with progressive cartilage destruction. And medicinal use. In particular, the present invention relates to the pharmacological partial use of chroman derivatives to treat or prevent osteoarthritis or rheumatoid arthritis.

  Osteoarthritis (OA, also called “degenerative joint disease”) is the most common rheumatic disease. The prevalence of this chronic disease increases with age, and this is a prominent cause of disability and poor quality of life among the elderly. The prevalence of this disease is higher in women than in men, and in women, the incidence of osteoarthritis increases after menopause. These observations suggest that estrogens may have affected cartilage metabolism and that decreased postmenopausal endogenous estrogen production increases the risk of subsequent OA progression. (D Felson & M Nevitt, 1998). Furthermore, most observational studies suggest that the use of estrogen after menopause is associated with a decrease in the occurrence of OA (Nevitt et al 1996). However, several studies have challenged this conclusion (Oliveria et al 1996, Erb et al. 2000, Nevitt et al 2001). Thus, so far, the potential role of estrogens in maintaining the integrity of the cartilage and in preventing OA progression has not been finally proven. A major component of the disease process in OA is irreversible cartilage destruction. Many years before the clinical diagnosis of the disease, the process is likely to have been initiated and continues until the end of the disease where almost all of the articular cartilage of the affected joint is lost . During this period, the mobility of the joint is greatly impaired and only joint replacement remains as a treatment option. Due to this long-term progression of pathological joint changes associated with OA, it is very difficult to elucidate factors involved early in the disease process. Rheumatoid arthritis (RA) is an inflammatory condition in which the articular cartilage of the affected joint is destroyed. The pathogenesis of RA is complex and numerous environmental and genetic factors have been suggested for its role in the progression of the disease. Furthermore, RA is more prevalent in women. Since estrogen has a positive function as an inhibitor of inflammation, it has been suggested that estrogen may also have a potentially beneficial effect on RA. Studies in human and animal models of OA and RA have demonstrated that macromolecule depletion of articular cartilage matrix progresses as the disease progresses. In OA, cartilage degeneration and osteophytes (abnormal bone twigs in affected joints) occur and progress on the bare parts of the joint bone. Symptoms of OA are joint pain, swelling and stiffness. In RA, cartilage destruction tends to occur more rapidly. The progression of joint destruction varies widely between individual patients, which is a marked cycle characterized by periods of high disease activity (rapidly occurring) with intermittent longer "silence" periods With a typical pattern. This periodic pattern of disease activity is particularly pronounced for RA.

  The most commonly used drugs to treat RA are more specific such as non-steroidal anti-inflammatory drugs (NSAIDs) and disease modifying anti-rheumatic drugs (DMARD's), and steroids or TNF-α antagonists Anti-inflammatory agent. NSAIDs and DMARD's also play an important role in the treatment of OA. The current treatment objective for these diseases is mainly to reduce pain and pathology. NSAIDs and DMARDs have been found to be effective in reducing the symptoms of OA and RA. However, the effect of suppressing cartilage catabolism has not been fully proven. Some of them (such as sodium salicylate) exhibit inhibitory properties on proteoglycan synthesis that may threaten the cartilage repair process. Other agents that do not inhibit proteoglycan synthesis (such as thiaprofenic acid) have shown in vitro that they can suppress OA cartilage catabolism (Jean-Pierre Pelletier et al., The Journal of Rheumatology 1989; 16: 5, 646-655). ). However, when administered to patients suffering from the latter disease, they failed to provide any significant protective effect on OA progression (Edward C. Huskisson et al., The Journal of Rheumatology 1995; 22: 10- 1941-1946). Doxycycline (which is a member of the tetracycline family) has also been shown in vivo to reduce metalloprotease activity while reducing the severity of OA lesions in a canine ACL model (Yu LP Jr et al., Arthritis Rheum). 35: 1150-1159, 1992). Recent data suggest that the action of corticosteroids is involved in the suppression of stromelysin-1 synthesis by chondrocytes (Pelletier et al., J Arthritis Rheum 37: 414-423, 1994). And Pelletier et al., J Lab Invest 72: 578-586, 1995).

  Compounds that act via estrogen and estrogen receptors in both OA and RA may have potentially beneficial effects. Selective alternatives to natural estrogens in hormone replacement therapy for treatments such as osteoporosis to avoid side effects such as breast cancer and endometrial growth while sustaining the beneficial effects of female hormones Estrogen receptor modulators (SERMs) have been used for several years. SERMs are synthetic compounds that exhibit some (but not all) effects of estrogen. For example, raloxifene is classified as SERM. This is because raloxifene suppresses bone loss and reduces serum cholesterol as an estrogen agonist, but acts as an estrogen antagonist by not stimulating the endometrial epithelium of the uterus. However, SERM's with strong estrogen agonist activity, for example on bone tissue, tend to be less selective for their antagonism on tissues such as endometrium. Therefore, because of undesirable side effects, many powerful SERM's have not been used as drugs in the prevention and treatment of osteoporosis and the like. An example of this is the chromane compound levormeloxifene as disclosed in US Pat. No. 3,822,287 and US Pat. No. 5,997,158.

  Progestins include steroid hormones related to progesterone, their derivatives and compounds having the effect of progesterone. They are widely used in hormone replacement therapy in combination with other steroid hormones to reduce (secondary) effects on endometrial epithelial growth. Recently, only progestins have been proposed for the treatment of bone related diseases as disclosed in WO 00/74684 and EPO 0474374. As disclosed in EPO 0665015, the combination of SERM raloxifene and progestin norethindrone acetate has been found to have a synergistic effect on osteoporosis by significantly improving bone mass index.

  While the beneficial effects of estrogen replacement therapy on the skeleton are evident, there are suggestions but no definitive evidence for the obvious effects of estrogen on OA or rheumatoid arthritis (Felson et al , 1998). However, SERM raloxifene has been demonstrated as an effective inhibitor of cartilage destruction in rodents (US Pat. No. 5,418,252). Recently, coumarin derivatives have been proposed as inhibitors of rheumatoid arthritis (US Pat. No. 6,291,456 and US Pat. No. 6,331,562). However, none of these compounds are recognized as clinically useful drugs for treating arthritis. Therefore, there is a continuing need to investigate the relationship between estrogen and OA, and the need to develop potent estrogen agonists, which can selectively target joint tissues that suffer from OA. And exhibit high bioavailability while minimizing side effects.

  The present invention provides, in the first aspect, droloxifene, levormeloxifene, chroman derivatives, nafoxidine, miproxyfen, arzoxifene, lasofoxifene, bazedoxifene, MDL-103323, EM-800, fulvest An effective amount of a selective estrogen receptor modulator (SERM) selected from Rant, ICI 183,780, ICI 164,384, 19-nortestosterone derivatives, and pharmaceutically acceptable esters, ethers and salts thereof A method of preventing, treating or alleviating symptoms including progressive cartilage destruction comprising administering to a subject.

  Preferably, the SERM comprises one or more of levormeloxifene, chroman derivatives, or 19-nortestosterone derivatives or pharmaceutically acceptable salts thereof.

  As used herein, “progestin or progestin agent” means a compound having progesterone activity (ie, inducing the formation of secretory endometrium), and specifically, norethindrone (also called norethisterone). ), Etinodiol, desogestrel, levonorgestrel, norgestrel, norgestimate, medroxyprogesterone, danazol, linoestrenol, didrogesterone, chlormadinone, promegesterone, gestrinone, algestone acetophenide, allylestrenol, cyproterone acetate, demegestone, Guestden, Osaterone, Hydroxyprogesterone hexanoate, Medrogestone, Megestrol, Nomegestrol, Ethynylnortestosterone, Norpregneno Emissions, NSC-9564, norethynodrel, de kiss norgestrel, gestodene, progesterone, chlormadinone acetate, drospirenone (dihydrospiro Lennon), or 3-keto Desolation have guest barrel or other compounds with salts and progestational activity. Progestin agents are well described in the art (see, for example, Martindale: The Extra Pharmacopoeia, 30th Edition, 1993, which is hereby incorporated by reference). A preferred salt is acetate. A more preferred progestin agent is norethindrone, and most preferred is norethindrone acetate. Progestin agents are also referred to as progestogens, gestagens or progesterone hormones.

  “Oosteoarthritis” or “OA” is a type of arthritis that is caused by the destruction of cartilage along with the resulting loss of cartilage of the joint. Cartilage is a proteinaceous material that serves as a “cushion” between joint bones. Osteoarthritis is also known as degenerative arthritis.

  “Rheumatoid arthritis” or “RA” is an inflammatory condition in which the articular cartilage of the affected joint is destroyed. The pathogenesis of RA is complex and numerous environmental and genetic factors have been suggested for its role in the progression of the disease.

  A “selective estrogen receptor modulator (SERM)” has some, but not all, of the estrogenic effects and exhibits activity as an agonist or antagonist of an estrogen receptor (eg, ER.α or ER.β). A synthetic compound shown in a tissue-dependent manner. Thus, as will be apparent to those skilled in the biochemical and endocrinology fields, the compounds of the present invention that function as SERMs are agonists of estrogen receptors in several tissues (eg bone, brain and / or heart). And can function as an antagonist in other types of tissues, such as the breast and / or inner wall of the uterus.

  As used herein, “chroman” means a 3,4-chroman derivative.

  As used herein, “coumarin” means a 2-chromen-2-one derivative.

  As used herein, a “cartilage specific biochemical marker” refers to a metabolite of cartilage catabolism that can be quantified in body fluids, which metabolite is an indicator of cartilage destruction throughout the body.

  According to a preferred method of the first aspect of the invention, the selective estrogen receptor modulator is selected from chroman derivatives. The claimed method may be carried out using any chroman derivative now known in the art or subsequently developed. For the synthesis of typical receptor antagonists, see only the examples of US Pat. No. 5,919,817, US Pat. No. 6,043,269, and EP 937057, EP 937060, and EP 937062. Which are hereby incorporated by reference in their entirety.

式中、
Ｒ¹はＨ、ＳＯ₂ＮＲ₂ ⁴、ＳＯ₂ＮＨＲ⁴、Ｃｌ、ＣＨ₃またはベンジルであり、
Ｒ²は、ＯＨ、ハロゲン、ニトロ、シアノ、ＳＨ、ＳＲ⁴、トリハロ−Ｃ₁−Ｃ₆アルキル、Ｃ₁−Ｃ₆アルキル、Ｃ₁−Ｃ₆アルコキシおよびフェニルからなる群より独立して選択される１〜５個の置換基で、必要に応じて置換されていてもよいフェニルであり、
Ｒ³は−−Ｘ−−（ＣＨ₂）_n−−Ｙで置換されたフェニルであり、式中、
Ｘは原子価結合、ＯまたはＳであり、
ｎは１〜１２の範囲の整数であり、
ＹはＨ、ハロゲン、ＯＨ、ＯＲ⁴、ＮＨＲ⁴、ＮＲ₂ ⁴、ＮＨＣＯＲ⁴、ＮＨＳＯ₂Ｒ⁴、ＣＯＮＨＲ⁴、ＣＯＮＲ⁴、ＣＯＯＨ、ＣＯＯＲ⁴、ＳＯ₂Ｒ⁴、ＳＯＲ⁴、ＳＯＮＨＲ⁴、ＳＯＮＲ₂ ⁴、ピロリジニル環であり、Ｈ、ＯＨ、ハロゲン、ニトロ、シアノ、ＳＨ、ＳＲ⁴、トリハロ−Ｃ₁−Ｃ₆アルキル、Ｃ₁−Ｃ₆アルキルおよびＣ₁−Ｃ₆アルコキシからなる群より独立して選択される１〜３個個の置換基で任意選択的に置換されていてもよく、
Ｒ⁴はＣ₁−Ｃ₆アルキルであり、
のクロマン誘導体ならびに、ＷＯ９８／１８７７３号に記載の光学異性体および幾何異性体、薬学的に許容可能なそのエステル、エーテルおよび塩からなる群より選択される。 In a preferred embodiment of the invention, the selective estrogen receptor modulator is of formula I

Where
R ¹ is H, SO ₂ NR ₂ ⁴ , SO ₂ NHR ⁴ , Cl, CH ₃ or benzyl;
R ² is independently selected from the group consisting of OH, halogen, nitro, cyano, SH, SR ⁴ , trihalo-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy and phenyl. 1 to 5 substituents, and optionally substituted phenyl,
R ³ is phenyl substituted with --X-(CH ₂ ) _n --Y, where
X is a valence bond, O or S,
n is an integer ranging from 1 to 12,
Y is H, halogen, OH, OR ⁴ , NHR ⁴ , NR ₂ ⁴ , NHCOR ⁴ , NHSO ₂ R ⁴ , CONHR ⁴ , CONR ⁴ , COOH, COOR ⁴ , SO ₂ R ⁴ , SOR ⁴ , SONR ⁴ , SONR ₂ ⁴ , a pyrrolidinyl ring, independent of the group consisting of H, OH, halogen, nitro, cyano, SH, SR ⁴ , trihalo-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl and C ₁ -C ₆ alkoxy Optionally substituted with 1 to 3 substituents selected from
R ⁴ is C ₁ -C ₆ alkyl;
And the optical isomers and geometric isomers described in WO 98/18773, pharmaceutically acceptable esters, ethers and salts thereof.

In another embodiment of the present invention there is provided the use of a compound of formula I, wherein centchroman (levormeloxifene) or (−)-3,4-trans-7-methoxy-2,2- Substituents R ² and R ³ are placed in trans, such as dimethyl-3-phenyl-4 {4 [2- (pyrrolidin-1-yl) ethoxy] phenyl} chroman.

In another preferred embodiment, the present invention provides compounds of formula I
Where
R ¹ is H, SO ₂ NR ₂ ⁴ , Cl, CH ₃ or SO ₂ NHR ⁴ ;
R ² and R ³ are arranged in cis,
R ² is independently selected from the group consisting of OH, halogen, nitro, cyano, SH, SR ₄ , trihalo-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl and C ₁ -C ₆ alkoxy. Is phenyl optionally substituted with ~ 3 substituents,
R ³ is phenyl substituted with --X-(CH ₂ ) _n --Y, where
X is a valence bond, O or S,
n is an integer ranging from 1 to 12,
Y represents H, OH, OR ⁴ , NHR ⁴ , NR ₂ ⁴ , NHCOR ⁴ , NHSO ₂ R ⁴ , CONH ⁴ , CONR ₂ ⁴ , COOH, COOR ⁴ , SO ₂ R ⁴ , SOR ⁴ , SONHR ⁴ , SONR ₂ ⁴ , A pyrrolidinyl ring, independently selected from the group consisting of H, OH, halogen, nitro, cyano, SH, SR ⁴ , trihalo-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl and C ₁ -C ₆ alkoxy Optionally substituted with 1 to 3 substituents,
R ⁴ is C ₁ -C ₆ alkyl;
And the use of optical and geometric isomers, pharmaceutically acceptable esters, ethers and salts thereof.

または、

の化合物の使用に関する。 In another preferred embodiment, the invention provides a compound of formula where R ¹ , R ² and R ³ are as defined above:

Or

To the use of the compound.

の化合物の使用に関する。 In another preferred embodiment, the invention provides a compound of formula wherein R ¹ is as defined above and m is an integer from 0-10.

To the use of the compound.

の化合物の使用に関する。 In another preferred embodiment, the invention provides that R ¹ is as defined above and R ⁶ represents one or more of the following substituents: methoxy, hydroxy, trifluoromethyl, fluoro and chloro formula:

To the use of the compound.

In a more preferred embodiment, the present invention provides the following compounds:
(+)-Cis-3- (4-fluorophenyl) -7-methoxy-4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+)-cis-3- (4-fluorophenyl) -7 -Methoxy-4- (4- (2-piperidinoethoxy) phenyl) chroman (+)-cis-3- (4-fluorophenyl) -7-methoxy-4- (4- (3-piperidinopropoxy) ) Phenyl) chroman (+)-cis-7-methoxy-3-phenyl-4- (4- (2-piperidinoethoxy) phenyl) chroman (+)-cis-7-methoxy-3-phenyl-4- (4- (3-Piperidinopropoxy) phenyl) chroman (+)-cis-7-methoxy-3- (4-methoxyphenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+ ) -Cis-7-meth Ci-3- (4-Phenyl-phenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+)-cis-7-methoxy-3- (4-methylphenyl) -4- (4 -(2-Pyrrolidinoethoxy) phenyl) chroman (+)-cis-7-methoxy-3- (4-methylphenyl) -4- (4- (2-piperidinoethoxy) phenyl) chroman (+)- Cis-7-methoxy-4- (4- (2-piperidinoethoxy) phenyl) -3- (4- (trifluoromethyl) phenyl) chroman (+)-cis-7-methoxy-4- (4- (2-Pyrrolidinoethoxy) phenyl) -3- (4- (trifluoromethyl) phenyl) chroman (+)-cis-7-methoxy-3- (3-methylphenyl) -4- (4- (2- Pyrrolidinoethoxy) phenyl) Roman (+)-cis-3- (3-fluorophenyl) -7-methoxy-4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+)-cis-7-methoxy-3- (3- Methoxyphenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+)-cis-7-methoxy-3- (3-methoxyphenyl) -4- (4- (2-piperidinoethoxy) ) Phenyl) chroman (+)-cis-7-methoxy-3- (3-methoxyphenyl) -4- (4- (3-piperidinopropoxy) phenyl) chroman (+)-cis-7-methoxy-4 -(4- (2-Pyrrolidinoethoxy) phenyl) -3- (3- (trifluoromethyl) phenyl) chroman (+)-cis-7-methoxy-4- (4- (2-piperidinoethoxy) Phenyl) -3- (3 -(Trifluoromethyl) phenyl) chroman (+)-cis-3- (2-fluorophenyl) -7-methoxy-4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+)-cis-7 -Methoxy-3- (2,3,4,5,6-pentafluorophenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+)-cis-7-methoxy-3- (2 , 3,4,5,6-pentafluorophenyl) -4- (4- (2-piperidinoethoxy) phenyl) chroman (+)-5- (cis-7-methoxy-3- (4-methylphenyl) ) -Chroman-4-yl) -2-methyl-benzoxazole (+)-cis-6-methoxy-3-phenyl-4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+)-cis- 6-methoxy-3- ( -Hydroxyphenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+)-cis-4- (4-hexylphenyl) -7-methoxy-3-phenylchroman (+)-cis-7 -Methoxy-3-phenyl-4- {4-2-2- (pyrrolidin-1-yl) ethoxy} phenyl} chroman (-)-cis-3- (4-fluorophenyl) -7-methoxy-4- (4- (2-Pyrrolidinoethoxy) phenyl) chroman (-)-cis-3- (4-fluorophenyl) -7-methoxy-4- (4- (2-piperidinoethoxy) phenyl) chroman (-)-cis -3- (4-Fluorophenyl) -7-methoxy-4- (4- (3-piperidinopropoxy) phenyl) chroman (-)-cis-7-methoxy-3-phenyl-4- (4- ( 2-pipe Jinoetokishi) phenyl) chroman (-) - cis -. sup. 7-methoxy-3-phenyl-4- (4- (3-piperidinopropoxy) phenyl) chroman (-)-cis-7-methoxy-3- (4-methoxyphenyl) -4- (4- (2 -Pyrrolidinoethoxy) phenyl) chroman (-)-cis-7-methoxy-3- (4-phenyl-phenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (-)-cis-7 -Methoxy-3- (4-methylphenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (-)-cis-7-methoxy-3- (4-methylphenyl) -4- (4 -(2-Piperidinoethoxy) phenyl) chroman (-)-cis-7-methoxy-4- (4- (2-piperidinoethoxy) phenyl) -3- (4- (trifluoromethyl) phenyl) Chroman (-)-shi -7-methoxy-4- (4- (2-pyrrolidinoethoxy) phenyl) -3- (4- (trifluoromethyl) phenyl) chroman (-)-cis-7-methoxy-3- (3-methylphenyl) ) 4- (4- (2-Pyrrolidinoethoxy) phenyl) chroman (-)-cis-3- (3-fluorophenyl) -7-methoxy-4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (-)-Cis-7-methoxy-3- (3-methoxyphenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (-)-cis-7-methoxy-3- (3-methoxy Phenyl) -4- (4- (2-piperidinoethoxy) phenyl) chroman (-)-cis-7-methoxy-3- (3-methoxyphenyl) -4- (4- (3-piperidinopropoxy) ) Phenyl) Roman (-)-cis-7-methoxy-4- (4- (2-pyrrolidinoethoxy) phenyl) -3- (3- (trifluoromethyl) phenyl) chroman (-)-cis-7-methoxy-4 -(4- (2-Piperidinoethoxy) phenyl) -3- (3- (trifluoromethyl) phenyl) chroman (-)-cis-3- (2-fluorophenyl) -7-methoxy-4- ( 4- (2-Pyrrolidinoethoxy) phenyl) chroman (-)-cis-7-methoxy-3- (2,3,4,5,6-pentafluorophenyl) -4- (4- (2-pyrrolidino (Ethoxy) phenyl) chroman (-)-cis-7-methoxy-3- (2,3,4,5,6-pentafluorophenyl) -4- (4- (2-piperidinoethoxy) phenyl) chroman ( -)-5- (cis-7-meth) Xyl-3- (4-methylphenyl) -chroman-4-yl) -2-methyl-benzoxazole (-)-cis-6-methoxy-3-phenyl-4- (4- (2-pyrrolidinoethoxy) Phenyl) chroman (-)-cis-6-methoxy-3- (3-hydroxyphenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (-)-cis-4- (4-hexylphenyl) ) -7-methoxy-3-phenylchroman (-)-cis-7-methoxy-3-phenyl 4- {4- {2- (pyrrolidin-1-yl) ethoxy} phenyl} chroman.
(+,-)-Cis-3- (4-fluorophenyl) -7-methoxy-4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+,-)-cis-3- (4-fluoro Phenyl) -7-methoxy-4- (4- (2-piperidinoethoxy) phenyl) chroman (+,-)-cis-3- (4-fluorophenyl) -7-methoxy-4- (4- ( 3-piperidinopropoxy) phenyl) chroman (+,-)-cis-7-methoxy-3-phenyl-4- (4- (2-piperidinoethoxy) phenyl) chroman (+,-)-cis- 7-methoxy-3-phenyl-4- (4- (3-piperidinopropoxy) phenyl) chroman (+,-)-cis-7-methoxy-3- (4-methoxyphenyl) -4- (4- (2-Pyrrolidinoethoxy) phenyl) chroma (+,-)-Cis-7-methoxy-3- (4-phenylphenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+,-)-cis-7-methoxy-3- (4-Methylphenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+,-)-cis-7-methoxy-3- (4-methylphenyl) -4- (4- (2 -Piperidinoethoxy) phenyl) chroman (+,-)-cis-7-methoxy-4- (4- (2-piperidinoethoxy) phenyl) -3- (4- (trifluoromethyl) phenyl) chroman (+,-)-Cis-7-methoxy-4- (4- (2-pyrrolidinoethoxy) phenyl) -3- (4- (trifluoromethyl) phenyl) chroman (+,-)-cis-7- Methoxy-3- (3-methylphenyl)- -(4- (2-pyrrolidinoethoxy) phenyl) chroman (+,-)-cis-3- (3-fluorophenyl) -7-methoxy-4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+,-)-Cis-7-methoxy-3- (3-methoxyphenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+,-)-cis-7-methoxy-3- (3-Methoxyphenyl) -4- (4- (2-piperidinoethoxy) phenyl) chroman (+,-)-cis-7-methoxy-3- (3-methoxyphenyl) -4- (4- ( 3-piperidinopropoxy) phenyl) chroman (+,-)-cis-7-methoxy-4- (4- (2-pyrrolidinoethoxy) phenyl) -3- (3- (trifluoromethyl) phenyl) chroman (+,-)-Cis- 7-methoxy-4- (4- (2-piperidinoethoxy) phenyl) -3- (3- (trifluoromethyl) phenyl) chroman (+,-)-cis-3- (2-fluorophenyl)- 7-methoxy-4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+,-)-cis-7-methoxy-3- (2,3,4,5,6-pentafluorophenyl) -4 -(4- (2-pyrrolidinoethoxy) phenyl) chroman (+,-)-cis-7-methoxy-3- (2,3,4,5,6-pentafluorophenyl) -4- (4- ( 2-piperidinoethoxy) phenyl) chroman (+,-)-5- (cis-7-methoxy-3- (4-methylphenyl) -chroman-4-yl) -2-methyl-benzoxazole (+, -)-Cis-6-methoxy-3-phenyl 4- (4- (2-pyrrolidinoethoxy) phenyl) chroman (+,-)-cis-6-methoxy-3- (3-hydroxyphenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) Chroman (+,-)-cis-4- (4-hexylphenyl) -7-methoxy-3-phenylchroman,
(+,-)-(Cis-4- (4- (2-pyrrolidinoethoxy) phenyl) -3- (4- (trifluoromethyl) phenyl) -chroman-7-yl) 2,2-dimethylpropano Art,
(+,-)-(Cis-4- (4- (2-pyrrolidinoethoxy) phenyl) -3- (3- (trifluoromethyl) phenyl) -chroman-7-yl) 2,2-dimethylpropano Art hydrochloride,
(+,-)-(Cis-3- (4-methylphenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) -chroman-7-yl) N, N-diethylcarbamate,
(+,-)-(Cis-3- (4-methylphenyl) -4- (4- (2-pyrrolidinoethoxy) phenyl) -chroman-7-yl) N, N-dimethylsulfamic acid ester,
(+,-) Cis 7-benzyloxy-4- (4-hydroxy-phenyl) -3-phenyl-chroman,
7-benzyloxy-4- [4- (4-chlorobutyloxy) -phenyl] -3-phenyl-chroman,
7-benzyloxy-4- [4- (2-chloroethyloxy) -phenyl] -3-phenyl-chroman,
7-benzyloxy-4- [4- (6-bromohexyloxy) -phenyl] -3-phenyl-chroman,
7-benzyloxy-4- [4- (10-bromodecyloxy) -phenyl] -3-phenyl-chroman,
(+,-)-Cis-3- (4-fluorophenyl) -7-methoxy-4- (4- (2-pyrrolidinoethoxy) phenyl) chroman,
(+-)-5- (cis-7-methoxy-3- (4-methylphenyl) -chroman-4-yl) -2-methyl-benzoxazole (+,-)-cis-6-methoxy-3- Phenyl-4- (4- (2-pyrrolidinoethoxy) phenyl) chroman,
(+,-)-Cis-4- (4-hexylphenyl) -7-methoxy-3-phenylchroman,
(+,-) Cis [4- (7-benzyloxy-3-phenyl-chroman-4-yl) -phenoxy] -acetic acid methyl ester,
(+,-) Cis 7-benzyloxy-4- [4- (6-morpholinohexyloxy) -phenyl] -3-phenyl-chroman,
(+,-) Cis 7-benzyloxy 4- [4- (6-dibutylaminohexyloxy) -phenyl] -3-phenyl-chroman,
(+,-) Cis 7-benzyloxy 4- [4- (dimethylaminodecyloxy) -phenyl] -3-phenyl-chroman,
(+,-) Cis 7-benzyloxy 4- [4- (morpholinodecyloxy) -phenyl] -3-phenyl-chroman,
(+,-) Cis 7-benzyloxy 4- [4- (2-dibutylaminoethyloxy) -phenyl] -3-phenyl-chroman,
(+,-) Cis 7-benzyloxy 4- [4- (butylaminoethyloxy) -phenyl] -3-phenyl-chroman,
(+,-) Cis 7-benzyloxy 4- [4- (morpholinoethyloxy) -phenyl] -3-phenyl-chroman,
(+,-) Cis 7-benzyloxy 4- [4- (N-methylpiperazinoethyloxy) -phenyl] -3-phenyl-chroman,
With respect to (+,-) cis 7-benzyloxy 4- [4- (morpholinobutyloxy) -phenyl] -3-phenyl-chroman, these pure optical isomers are also included.

（−）−シス−｛３−（４−トリフルオロメチルフェニル）−４−［４−（２−ピロリジン−１−イル−エトキシ）−フェニル］｝−７−メトキシクロマン

（−／＋）−シス−｛３−（３−メトキシフェニル）−４−［４−（２−ピロリジン−１−イル−エトキシ）−フェニル］｝−７−メトキシクロマン

（−／＋）−シス−｛３−（３−ヒドロキシフェニル）−４−［４−（２−ピロリジン−１−イル−エトキシ）−フェニル］｝−クロマン−７−オール

（−／＋）−シス−｛３−（４−トリフルオロメチルフェニル）−４−［４−（２−ピロリジン−１−イル−エトキシ）−フェニル］｝−クロマン−７−オール

（−）−シス−７−メトキシ−３−フェニル−４−（４−（２−ピロリジノエトキシ）−フェニル）クロマン

（＋／−）−シス−７−メトキシ−３−（３−メトキシフェニル）−４−（４−（２−ピロリジノエトキシ）−フェニル）クロマン

（＋／−）−シス−７−メトキシ−３−（４−トリフルオロメチルフェニル）−４−（４−（２−ピロリジノエトキシ）−フェニル）クロマン

および
（−）−シス−７−メトキシ−３−（４−トリフルオロメチルフェニル）−４−（４−（２−ピロリジノエトキシ）−フェニル）クロマン

に関する。 In the most preferred embodiment, the present invention provides the following compounds:
(−)-Cis- {3-phenyl-4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-chroman-7-ol

(−)-Cis- {3- (4-trifluoromethylphenyl) -4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-7-methoxychroman

(− / +)-Cis- {3- (3-methoxyphenyl) -4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-7-methoxychroman

(− / +)-Cis- {3- (3-hydroxyphenyl) -4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-chroman-7-ol

(− / +)-Cis- {3- (4-trifluoromethylphenyl) -4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-chroman-7-ol

(-)-Cis-7-methoxy-3-phenyl-4- (4- (2-pyrrolidinoethoxy) -phenyl) chroman

(+/-)-cis-7-methoxy-3- (3-methoxyphenyl) -4- (4- (2-pyrrolidinoethoxy) -phenyl) chroman

(+/-)-cis-7-methoxy-3- (4-trifluoromethylphenyl) -4- (4- (2-pyrrolidinoethoxy) -phenyl) chroman

And (-)-cis-7-methoxy-3- (4-trifluoromethylphenyl) -4- (4- (2-pyrrolidinoethoxy) -phenyl) chroman

About.

  The present invention includes the use of a SERM as defined above for preparing a medicament for use in preventing, treating or alleviating symptoms including progressive cartilage destruction.

In another embodiment, the present invention provides:
1) a selective estrogen receptor modulator (SERM) or a pharmaceutically acceptable salt thereof;
2) preventing symptoms including progressive cartilage destruction, comprising administering to a subject an effective amount of a combination of progestins or a pharmaceutically acceptable salt thereof in combination that reduces cartilage destruction, It relates to a method of treating or alleviating.

  The present invention further includes (1) a chroman derivative or a coumarin derivative or a pharmaceutically acceptable salt or solvate thereof, (2) a progestin or a salt thereof, and another compound having progesterone activity. A pharmaceutical preparation for treating arthritis, which is contained together with a pharmaceutically acceptable carrier in an amount that suppresses OA or RA.

  In particular, the present invention relates to raloxifene, droloxifene, tamoxifen, 4-hydroxy-tamoxifen, 4′-iodotamoxifen, toremifene, (deaminohydroxy) toremifene, clomiphene, levormeloxifene, chroman derivatives, coumarin derivatives, idoxifene, Nafoxidine, Myproxyfen (TAT-59), Arzoxifene (LY-353381), Lasofoxifene (CP-336156), MDL-103323, EM-800, Fulvestrant (ICI-182, 780) ICI 183 780, ICI 164,384, diethylstilbestrol, genistein, nafoxidine, GW 5638, panomiphene, nitromiphene citrate, moxesterol, diphenol hydrochloride Sene, erythro-MEA, allenolic acid, cyclophenyl, chlorotrianicene, ethamoxytriphetol, triparanol 19-norprogesterone derivatives, 19-nortestosterone derivatives and pharmaceutically acceptable esters, ethers and salts A selective estrogen receptor modulator (SERM) selected from the group consisting of: norethindrone, ethinodiol, desogestrel, levonorgestrel, norgestrel, norgestimate, medroxyprogesterone, danazol, linoestrenol, didrogesterone, chlormadinone, promegesterone, guest Linone, Algestone acetophenide, Allylestrenol, Cyproterone acetate, Demegestone, Guestden, Osaterone, Hexanoic acid hydride Roxyprogesterone, medrogestone, megestrol, nomegestrol, ethynylnortestosterone, norpregnenolone, NSC-9564, norethinodrel, dexnorgestrel, guestden, progesterone, chlormadinone acetate, drospirenone (dihydrospirorenone), or 3-keto Treating arthritis comprising administering to a subject an effective amount of a combination of desogestrel and optical and geometric isomers, progestins selected from the group consisting of pharmaceutically acceptable esters and salts thereof On how to do or alleviate.

  Preferred SERM's include droloxifene, tamoxifen, levormeloxifene, chroman derivatives, nafoxidine, toremifene, TAT-59, LY-353381, CP-336156, MDL-103323, EM-800, ICI-182, ICI 183,780 and 19-nortestosterone derivatives or pharmaceutically acceptable salts thereof.

  This aspect of the invention combines the side effects of hormone replacement therapy (eg, increased breast cancer incidence and endometrial epithelial growth, etc.), particularly with specific selective estrogen receptor modulators (SERMs) alone, combined with progestins It is related with observation that it can suppress by administering. When treating, preventing, or alleviating diseases associated with progressive cartilage destruction such as osteoarthritis and rheumatoid arthritis, it is essential to administer a long-term effective inhibitor of cartilage destruction. is there. Thus, even minor side effects of the inhibitor may have a dramatic impact on patient health over time and should be avoided or minimized to improve the value of the patient's life. And the cost of insurance schemes for costly complications should be saved.

  According to a preferred method of the second aspect of the invention, the selective estrogen receptor modulator is selected from chroman derivatives.

  Preferred chroman derivatives are as described above.

  In another preferred embodiment of the invention, the progestin is progesterone, norethindrone acetate, ethinodiol, desogestrel, levonorgestrel, norgestrel, norgestimate, medroxyprogesterone and esters thereof or a pharmaceutically acceptable salt thereof.

  In a further preferred embodiment of the invention, the progestin is norethindrone or norethindrone acetate and esters or pharmaceutically acceptable salts thereof.

  In another aspect, the invention is directed to inhibiting estrogen production, wherein the composition comprises a 3,4 chroman or coumarin derivative or a pharmaceutically acceptable salt thereof and a progestin or pharmaceutically It relates to a pharmaceutical composition for use in the therapeutic or prophylactic treatment of a disease, comprising an effective amount of a combination of acceptable salts thereof and a pharmaceutical carrier or diluent.

  In one embodiment, the present invention provides breast cancer, testicular cancer, osteopenia or bone loss leading to osteoporosis, endometriosis, heart disease, hypercholesterolemia, enlarged prostate, prostate cancer, obesity, hot flash, skin effect To treat or prophylactically treat symptoms such as mood swings, memory loss, urinary incontinence, hair loss, cataracts, natural hormone upset, and adverse reproductive toxicity associated with exposure to environmental chemicals It relates to the use of the pharmaceutical composition for treatment.

  In a preferred embodiment, the present invention is based on progressive cartilage destruction, wherein the composition comprises a 3,4 chroman or coumarin derivative or a pharmaceutically acceptable salt thereof and a progestin or a pharmaceutically acceptable The present invention relates to a pharmaceutical composition for use in therapeutic or prophylactic treatment of diseases, comprising an effective amount of a combination thereof and a pharmaceutical carrier or diluent.

  In a further preferred embodiment, the invention relates to progressive cartilage destruction, wherein the composition comprises a 3,4 chroman or coumarin derivative or a pharmaceutically acceptable salt thereof and norethindrone or norethindrone acetate or pharmaceutical Relates to a pharmaceutical composition for use in therapeutic or prophylactic treatment of diseases comprising an effective amount of a combination of pharmaceutically acceptable salts thereof and a pharmaceutical carrier or diluent.

  In a most preferred embodiment, the invention is based on progressive cartilage destruction, wherein the composition comprises an effective amount of cis-3,4 diarylchroman or a pharmaceutically acceptable salt thereof and norethindrone or acetic acid. A pharmaceutical composition for use in therapeutic or prophylactic treatment of a disease, comprising a combination of an effective amount of norethindrone or a pharmaceutically acceptable salt thereof and a pharmaceutical carrier or diluent. .

  In a preferred embodiment of the invention, the effective amount of the selective estrogen receptor modulator or pharmaceutically acceptable salt thereof per person per day is 0.01 to about 100 mg, 0.1 to about 10 mg per day is preferred, with 0.2 to about 1.0 mg per individual per day being most preferred.

  In another preferred embodiment of the invention, the effective amount of the progestin or pharmaceutically acceptable salt thereof and the pharmaceutical carrier or diluent per person per day is 0.01 to about 100 mg, 0.1 to about 10 mg per person per day is preferred, and 0.2 to about 1.0 mg per person per day is most preferred.

  In a most preferred embodiment of the invention, an effective amount of the progestin or a pharmaceutically acceptable salt thereof and the pharmaceutical carrier or diluent are used to significantly reduce the estrogenic side effects induced by SERMs. It is a sufficient amount.

  In yet another preferred embodiment, the present invention relates to a pharmaceutical composition as described above, which is in the form of an oral unit dose or a parenteral unit dose.

  In a further preferred embodiment, the present invention relates to the administration of a therapeutically effective amount of a combination of Compound I and progestin to a mammal in need thereof.

  In a most preferred embodiment, the invention relates to administering a therapeutically effective amount of a combination of Compound I and norethindrone or norethindrone acetate to a human in need thereof.

  In another embodiment, the present invention provides osteoarthritis or rheumatoid arthritis comprising administering to a human in need thereof a combination of a therapeutically effective amount of Compound (I) and progestin. Relates to a method of treating.

  In another aspect, the present invention relates to the use of cartilage specific biochemical markers for titrating the above doses. The therapeutically effective dose titration is based on quantifying one or more of the above cartilage matrix protein fragments. Various doses of the pharmaceutical composition are given to the mammal for at least 2 to 4 weeks, and the cartilage destruction response is evaluated for each dose to select the minimal therapeutically effective dose. A therapeutically effective dose is defined as a dose that significantly reduces certain biomarkers of cartilage.

  In a preferred embodiment, the therapeutically effective dose results in a 20-90% reduction in the control level of a particular biomarker of cartilage, more preferably a 30-70%, particularly preferably a 40-50% reduction. Bring.

  In a preferred embodiment, the markers are described in Christgan et al. , 2001, a CartiLaps ELISA.

  In yet another aspect, the invention relates to droloxifene, tamoxifen, levormeloxifene, chroman derivatives, coumarin derivatives, idoxifene, nafoxidine for preventing, treating or alleviating symptoms including progressive cartilage destruction , Toremifene, TAT-59, LY-353381, CP-336156, MDL-103323, EM-800, ICI-182, ICI 183,780 and 19-nortestosterone derivatives or pharmaceutically acceptable salts thereof It relates to the use of a composition comprising a selective estrogen receptor modulator (SERM).

  In another embodiment, the present invention relates to droloxifene, tamoxifen, levormeloxifene, chroman derivatives, coumarin derivatives, idoxifene, nafoxidine, toremifene, TAT-59, LY-353381, lasofoxifene, MDL-103323, Selective selected from EM-800, ICI-182, ICI 183,780, ICI 164,384, ICI 183,780, GW 5638, 19-nortestosterone derivatives and pharmaceutically acceptable esters, ethers and salts thereof The present invention relates to a method for preventing, treating or alleviating a symptom including progressive cartilage destruction, comprising a step of administering to a subject an amount of estrogen receptor modulator (SERM) that inhibits cartilage destruction.

  In preferred embodiments, the present invention is directed to droloxifene, levormeloxifene, chroman derivatives, nafoxidine, TAT-59, LY-353381, lasofoxifene, MDL-103323, EM-800, ICI-182, 780, A selective estrogen receptor modulator (SERM) selected from ICI 183,780, ICI 164,384, GW 5638, ICI 183,780, 19-nortestosterone derivatives and pharmaceutically acceptable esters, ethers and salts thereof. It relates to a method for preventing, treating or alleviating symptoms including progressive cartilage destruction, comprising administering an effective amount to a subject.

で示されるコア構造を含むＳＥＲＭ’Ｓであることが理解できるだろう。 According to each embodiment of the present invention, preferred SERM's are those represented by the following formula 2:

It will be understood that the SERM'S includes a core structure represented by

  The dotted bond indicates a single bond or a double bond.

中の印の付いたＲの位置で拡張してもよい。 It will be appreciated that this is included in chroman and in many other (but not all) known SERM's. In particular, the core structure is represented by the following formula 3:

You may expand at the position of R marked inside.

  Each extension at the R position can be any structure that binds to the core structure of Formula 2, thereby producing a SERM.

  Preferably, the binding affinity Ki for estrogen receptor α of SERMs for use in any aspect of the invention should be 10 μM or higher, preferably 1 μM or higher and / or binding affinity for estrogen receptor β. Is 10 μM or more, preferably 1 μM or more. In a most preferred embodiment of the invention, the SERM comprises levormeloxifene, a chroman derivative and a 19-nortestosterone derivative or a pharmaceutically acceptable salt thereof.

The common names of the compounds cited above are more fully defined as follows.
Olmeroxifene trans-1- (2- (4-(-methoxy-2,2-dimethyl-3-phenyl) -5H-benzopyran-4-yl) -phenoxy) ethylyl) -pyrrolidenraloxifene / Evista LY117081- Hydroxy-2- (4-hydroxyphenyl) -benzo (p) -thien-3-yl-4- (2- (1-piperidinyl) -ethoxy) -phenylmethanetoremifene / fareston FC-1157 / NK622 Z-2- (4- (1,2-Diphenyl-1--4-chlorobutenyl) -phenoxy) -N, N-dimethylethanamine tamoxifen ICI 46,474 Z-2- (4- (1,2-diphenyl-1-butenyl) -Phenoxy) -N, N-dimethylethanamine arzoxifene LY-353381 6-H Droxy-3- (4- (2- (1- (piperidinyl) -ethoxy) -phenoxy) -2- (4-hydroxyphenyl) -benzo (p) -thiophenlasofoxifene CP 336,156 cis-1R- [ 4'-pyrrolidino-ethoxyphenyl] -2S-phenyl-6-hydroxy-1,2,3,4, -tetrahydronaphthalene D-(-)-tartrate bazedoxifene ERA-923 / TSE-424 1H-indole -5-ol 1-[[4- [2- (hexahydro-1H-azepin-1-yl) ethoxy] phenyl] methyl] -2- (4-hydroxyphenyl) -3-methyl-monoacetate tesmilifene DPPE N, N-diethyl-2- [4- (phenylmethyl) -phenoxy] ethanamine; ICI 164,384 Nn-butyl Ru-N-methyl-11- (3,17b-dihydroxyosla-1,3,5 (10) -trien-7a-yl) undecanamido-osmepifen FC-1271a Z-2- (4- (4- Chloro-1,2-diphenyl-but-1-enyl) phenoxy) ethanol; CHF-4056 3-phenyl-4-[[4- [2- (1-piperidinyl) ethoxy] phenyl] methyl] -2H-1- Benzopyran-7-olmiproxyfen TAT-59 (E) -4- [1- [4- [2- (dimethylamino) ethoxy] -phenyl] -2- (4-isopropyl) phenyl-1-butenyl] -phenyl -Monophosphate Idoxyphene SB 223 030 Pyrrolidino-4-iodotamoxifen Faslodex ICI 182,780 7a- (9- ( 4,4,5,5,5-pentafluoro-pentyl-sulfinyl) -nonyl) -estradi-1,3,5 (10) -triene-3,17b-diol.
Nafoxidine 11100A 1- (2- (4- (3,4-dihydro-6-methoxy-2-phenyl-naphthyl) -phenoxy) -ethyl) -pyrrolidine; MDL-103323 (E) -1-butanamine, 4- [ 4- (2-Chloro-1,2-diphenylethenyl) phenoxy] -N, N-diethyl-dihydrogen citrate; EM-800 / SCH57050 (s)-(+)-4- [7- (2,2 -Dimethyl-1-oxopropoxy) -4-methyl-2- [4- [2- (1-piperidyl) ethoxy] phenyl] 2H-1-benzopyran-3-yl] -phenyl 2,2-dimethylpropanoate Droloxifene EM-652 / SCH57058 Z-2- (4- (1-3-phenol-2-phenyl-1-butenyl) -phenoxy) -N, N- Methylethanamine 7-hydroxy-2- (4- (2-piperidinoethoxy) phenyl) -3- (4-hydroxyphenyl) -4-methyl-2H-1-benzopyranpanomiphene GYKI 13504 (E) -1,2, -diphenyl-1- [4- [2 (2-hydroxyethylamino) -ethoxy] -phenyl] -3,3,3-trifluoropropene clomiphene 2- (4- (2-chloro-1 , 2-diphenylethenyl) phenoxy-N, N-diethylethanolamine gindoxifene 1H-indole-5-ol-2- (4-acetoxy) phenyl) -1-ethyl-3-methylacetate; GW 5638 3- [4- (1,2-Diphenylbut-1-enyl) phenyl] acrylic acid

  The compounds of the present invention may be prepared by utilizing the chroman chemistry well known in the art. For example, P.I. K. Arora, P.A. L. Kole and S.K. Ray, Indian J. et al. Chem. 20B, 41-5, 1981; Ray, P.M. K. Grover and N.M. Anand, Indian J. et al. Chem. 9, 727-8, 1971; Ray, P.M. K. Grover, V.M. P. Kamboj, S .; B. Betty, A.M. B. Kar and N.C. Anand, J .; Med. Chem. 19, 276-9, 1976; Md. Salman, S.M. Ray, A .; K. Agarwal, S.A. Durani, B.D. S. Betty, V.M. P. Kamboj, and N.J. Anand, J .; Med. Chem. 26,592-5, 1983; Sim, K .; Bull. Singapore Natl. Inst. Chem. 22, 69-74, 1994.

  However, the present invention further relates to general methods for preparing compounds of formula (I) as described in US Pat. No. 6,043,269.

[Pharmaceutical preparation]
The invention further relates to a pharmaceutical composition comprising an effective amount of a compound according to the invention and a pharmaceutical carrier or diluent. The form of such a composition is preferably an oral unit dose or a parenteral unit dose. The compounds according to the invention may be prepared for administration by any route consistent with their pharmacokinetic properties. Compositions that can be administered orally include tablets, capsules, powders, granules, electuary, liquid or gel formulations, such as oral, topical or sterile parenteral solutions or suspensions, etc. It may be in the form of Tablets and capsules for oral administration may be in the form of a unit dose, commonly used excipients such as syrup, acacia, gelatin, sorbitol, tragacanth or polyvinylpyrrolidone binders; lactose, Bulking agents such as sugar, corn starch, calcium phosphate, sorbitol or glycine; lubricants for tablets such as magnesium stearate, talc, polyethylene glycol or silica; disintegrants such as potato starch; or acceptable wetting agents such as sodium lauryl sulfate , May be included. The tablets may be coated according to methods well known in standard pharmaceutical practice. Oral liquid formulations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or presented as a dry product for reconstitution with water or other suitable medium prior to use May be. For such liquid preparations, commonly used additives such as suspending agents such as sorbitol, syrup, methylcellulose, glucose syrup, gelatin hardened edible fat; emulsifiers such as lecithin, sorbitan monooleate, or acacia Non-aqueous media such as tonsils, fractionated coconut oil, glycerin, propylene glycol or non-aqueous media such as ethyl alcohol (may include edible oils); such as methyl p-hydroxybenzoate or propyl hydroxybenzoate or sorbic acid Preservatives may be included, and perfume or colorant commonly used may be included as required.

  A unit dose for oral administration may contain about 0.01-100 mg of the compound of the present invention, preferably about 0.1-10 mg, most preferably about 0.2-1 mg. The appropriate daily dose for a mammal can vary widely depending on the condition of the patient. However, the dose of the compound of general formula I is about 0.1-10 mg / kg body weight, and about 0.2-1 mg / kg body weight would be more reasonable.

  Creams, lotions or ointments may be made from the drug for topical application to the skin. Cream or ointment formulations that may be used for drugs are commonly used formulations that are well known in the art and are described, for example, in standard textbooks of pharmacology such as the British Pharmacopoeia.

  The active ingredient may be administered parenterally in a sterile medium. Depending on the medium and the concentration used, the drug may be suspended or dissolved in the medium. Adjuvants such as local anesthetics, preservatives and buffering agents can be conveniently dissolved in the medium.

  Drugs for intravaginal administration include, in addition to the commonly used vaginal suppositories, soft capsules containing liquid oily bases or ointments, and the same, for use in injections A tube may be mentioned. Examples of the base include an oleaginous base, a water-soluble base, an emulsion base, and an ointment-like base. Specifically, as the oleaginous base, peanut oil, olive oil, corn oil, castor oil, cacao oil, lauric fat, glycerin fatty acid ester, specifically Pharmasol (a product of NOF), Witepsol (Damitite Nobel) Products), SB-base (product of Kaneka), and lanolin fats and oils. Specific examples of the water-soluble base include polyethylene glycol, polypropylene glycol, glycerin and glycero gelatin. The emulsion base is a base obtained by emulsifying a water-soluble base and an oleaginous base, and may be an O / W type or a W / O type. In the present invention, an oleaginous base is preferably used, and thereby the stability of PG is increased and the excellent pharmacological effect of PG can be achieved. These bases may be used alone or in appropriate combination. The compounds of the present invention may be incorporated into implants, vaginal rings, patches, gels, and any other sustained release formulation.

  Nasal administration from solution as a spray nasal spray may be applied and may be dispensed as a spray by a variety of methods known to those skilled in the art. A preferred system for dispensing liquid as a spray is disclosed in US Pat. No. 4,511,069. Such a system was used when performing the operations described in the examples described below. Such spray nasal spray solutions include the drug, ie, the drug to be delivered, a nonionic surfactant to facilitate absorption of the drug, polysorbate 80, and one or more buffering agents. In some embodiments of the invention, the spray nasal solution further comprises a propellant. The pH of the spray nasal spray solution is preferably between pH 6.8 and 7.2.

  The following examples of the invention are presented for purposes of illustration and not limitation.

Ovariectomy induces cartilage erosion.
Several animal models of osteoarthritis have been described in the literature (Bendele et al., 2001). Naturally occurring OA is found in some animal species such as mice, golden hamsters, guinea pigs and non-human primates. In addition, various transgenic mouse species that are prone to OA have been developed, and surgically induced joint damage has also been extensively studied. For slowly progressive forms of human disease, a model in which the disease progresses is probably most appropriate. These models show the importance of factors during OA progression, such as the role of matrix metalloproteinases (MMPs) during cartilage destruction, the importance of TGF-β during osteophyte formation, and the production of MMPs and chondrocytes It is very useful in identifying the role of nitric oxide in the induction of apoptosis. However, at present, these models have not been applied to study the potential role of estrogens in maintaining the integrity of the cartilage. Recent studies in cynomolgus monkeys revealed that OA-like pathological changes occurred in the joints of animals from which the ovaries were removed, and that this change could be prevented by exogenous estrogen preparations ( Ham et al 2002). This suggests that compounds that act on estrogen receptors such as estrogens and SERMs potentially have a role in protecting cartilage.

  We have established a model for assessing postmenopausal OA in vivo by performing ovariectomy (OVX) in female rats. The validity of this model was evaluated by quantifying articular cartilage damage in 5 and 7 month old rats. Here, the quantification of the injury was made by histological calculations using a scoring system developed to quantify erosive changes in the knee joint. In addition, cartilage and bone metabolic levels were assessed by specific biochemical markers. Cartilaps (CTX-II) specific to degradation products of type II collagen is an index of cartilage turnover (Christgan et al 2001), and RatLaps (CTX-I) is a type I collagen degradation derived from bone resorption. Specific to the product.

  The animals in this study consisted of two populations of 20 female Sprague-Dawley rats (obtained from Charles River laboratory, Germany). One population was 5 months old and the second population was 7 months old. Each group was divided into two groups. Each group had 10 animals and was raised for 9 weeks. Two months before the start of the study, animals were received at the animal facility so that they could acclimatize to the environment. Body weight was measured at the baseline and the animals were randomized and divided into two groups. All rats are ovariectomized by first anesthetizing with Hypnorm-Dormicum (1 part of Hypnorm® + 1 part of Dormicum + 2 parts of sterile deionized water). Rats are given 0.15-0.2 mL / 100 g body weight and standard sham or ovariectomy is performed. Animals were weighed weekly for the entire duration of the experiment. Throughout the study, animals were weighed at regular intervals and blood samples as well as urine samples were collected. Collect urine samples as spots of urine samples by gently urinating the animal's abdomen or by collecting the urine samples obtained by placing the animals in a metabolic measurement cage for 30-60 minutes did. At the end, the knee was separated and histologically analyzed for cartilage erosion.

  The knee was decalcified with 10% formic acid and 2% formaldehyde. The decalcified knee joint was cut into two sections along the medial collateral ligament and embedded in paraffin. The embedded knee was then cut from the medial collateral ligament to three different depths (0, 250, and 500 μm), and each section was then stained with toluidine blue. The cartilage sections were scored for the following blinded parameters: erosion, proteoglycan loss, chondrocyte blackening / death, chondrocyte loss, cysts, fibrillation, and bone formation in cartilage. All changes were measured as a percentage of the total surface of the cartilage, and the incidence of erosion in the (OVX / sham surgery) group was also calculated. RatLaps (CTX-I, bone resorption) and CartiLaps (CTX-II, cartilage destruction) levels of each of the serum and urine samples were measured.

Urinary Cartilaps ELISA is measured using a competitive ELISA method that utilizes a monoclonal antibody (MabF46) specific for the C-telopeptide fragment of type II collagen (Christgan et al 2001). First, a biotinylated peptide derived from a C-telopeptide of type II collagen (EKGPDP) was incubated on a streptavidin microtiter plate and then evaluated by adding the sample as well as the primary antibody. . A monoclonal antibody MabF46 specific for the C-telopeptide fragment of type II collagen was used in a competitive ELISA format developed for the measurement of urine samples. In summary, the evaluation was performed as follows. A biotinylated peptide derived from C-telopeptide of type II collagen (EKGPDP) was diluted with a coating buffer (1.5 mg / L), and 100 μL thereof was added to a streptavidin-coated microtiter plate ( Micro-Coat, Munich, Germany) was pipetted into each well. Plates were incubated at 18-22 ° C. for 30 ± 5 minutes. The plate was washed 5 times with wash buffer and 40 μL of standard, control or unknown sample was pipetted into the wells. All samples were measured in duplicate. 100 μL of primary antibody (MabF46) was diluted with assay buffer to a concentration of 19 mg / L and pipetted into each well. After overnight incubation at 4 ° C., the plates were washed and peroxidase labeled anti-mouse antibody was added, and then the antibody bound to the chromogenic peroxidase substrate was visualized. The absorbance at 450 nm (650 nm is a reference) was measured with an ELISA plate reader, and the concentration of the unknown sample was determined by preparing a standard curve obtained from measurement of a standard substance using a known type II collagen peptide. The concentration of CartiLaps ELISA [ng / L] was normalized to the total amount of urine creatinine [mmol / L]: concentration / creatinine [ng / mmol]. The measurement accuracy of this evaluation method was 7.1% and 8.4% with respect to the deviation between evaluations and during evaluation, respectively.

C-telopeptide degradation products of type I collagen are measured in a RatLaps ELISA using a specific monoclonal antibody in a competitive ELISA method. This evaluation method is suitable for both urine and serum samples. However, only serum samples were evaluated in this study. A commercially available product is available for this evaluation (Nordic Bioscience Diagnostics, Herreb, Denmark). All serum samples measured by this evaluation method were derived from animals. The animals were fasted for at least 6 hours prior to sampling. Briefly, this evaluation is performed by incubating a biotinylated form of a synthetic peptide representing the EKSQDGGR epitope of the C-telopeptide. This is followed by the addition of sample and primary antibody, and after overnight incubation, the amount of bound antibody is visualized using a secondary antibody labeled with peroxidase and a chromogenic peroxidase substrate. The concentration in the sample was determined from the result of creating a calibration curve based on the measurement result of the standard synthetic peptide.

  In both populations, erosion inside the femur progressed significantly in the OVX group compared to the sham-operated group (p = 0.037 and p = 0.08 for 5 and 7 month old animals, respectively). . OVX showed further erosion of cartilage in other areas of the knee joint. However, the difference reached statistical significance only inside the tibia of the 5 month old population and outside the femur of the 7 month old population (p <0.01 and p = 0.009, respectively). The total score across the four areas showed that erosion in the OVX group was significantly more advanced than that in the sham-operated group in both 5 month and 7 month old animals (p = 0.0, respectively). 09 and p = 0.008) (FIG. 1).

In both 5 month and 7 month old animals, OVX led to very significant increases in CTX-I and CTX-II levels (FIG. 2). This result is consistent with histological data. Bone resorption measured using the CTX-I marker continues to increase throughout the 9-week study period in OVX animals, whereas the increase in CartiLaps becomes most evident after 4 weeks of OVX and then decreases Is shown. The correlation between changes in CartiLaps and erosion was evaluated (Figure 3). A correlation coefficient R ² = 0.27 indicates that there is a significant (p = 0.039) correlation between the two parameters, which could be expected. The correlation of CartiLaps to erosion outside the femur (where the most severe erosion was seen) was also significant (R ² = 0.25, p = 0.048; data not shown).

  Another method for assessing the association between CartiLaps and erosion quantified by histology assesses not only changes in CartiLaps levels, but also the association between levels and subsequent cartilage erosion It is to be. In FIG. 4, data for a 7 month old population was processed in this manner. According to CartiLaps 4 weeks after OVX, the animals were divided into two groups: small value CartiLaps (28-75 ng / mmol) and large value (97-520 ng / mmol). Animals showing small values of CartiLaps data have only slight erosion, whereas animals showing large values of CartiLaps not only have a high incidence of erosion, but also have a high percentage of erosion (FIG. 4). Similar results were obtained when analyzing data from 5 month old animals.

  In conclusion, the results demonstrate that a significant increase in both bone and cartilage turnover is induced in OVX-treated rats and that cartilage erosion in OVX-treated rats appears after 9 weeks.

  This suggests that compounds acting through estrogens and estrogen receptors have an important role in maintaining normal cartilage turnover and, as demonstrated in the following examples, It suggests that estrogen and SERM's may protect cartilage. Furthermore, the results confirm that it is reasonable to use the OVX model as an in vivo model for post-menopausal OA. Finally, the results demonstrate that the CartiLaps assessment may be used as a valid biochemical marker for subsequent articular cartilage destruction in a rat model of arthritis.

Levormeloxifene is a potent inhibitor of cartilage destruction in rats from which the ovaries have been removed.
In order to test the potential cartilage protective effect of SERM, levormeloxifene was evaluated in an OVX model of postmenopausal OA (see Example 1). Seventy six month old female Sprague-Dawley rats were obtained from Charles River, Germany and used in this study. Ten animals were sham operated and the rest (60 animals) were ovariectomized. Animal ovariectomy and handling were performed as described in Example 1. OVX animals are divided into 5 groups of 12 animals, vehicle, 0.1 mg / kg / day 17α-ethynylestradiol (Sigma, E-8875, lot number 21K1267) or triplicate of levormeloxifene (0.2, 1 or 5 mg / kg / day). All treatments were performed orally in suspension (50% propylene glycol (Unikem, Copenhagen, Denmark, catalog number 291443), 0.075 M NaCl). Conducted daily 5 days a week and not administered on weekends. Throughout the duration of the study, animals were weighed at regular intervals and blood / urine samples were collected. Collect urine samples as spots of urine samples by gently urinating the animal's abdomen or by collecting the urine samples obtained by placing the animals in a metabolic measurement cage for 30-60 minutes did. At the end, the knee was separated and histologically analyzed for cartilage erosion.

  At the lowest dose (0.2 mg / kg / day, administered 5 days per week), the development of body weight as sham-operated or OVX-treated animals is the same, ie a study period of 9 weeks During this period, there was no significant change in body weight. On the other hand, the medium dose (1 mg / kg / day, administered 5 days a week) and the high dose (5 mg / kg / day, administered 5 days a week) may grow slightly slower than these reference groups. (Data not shown).

  Levormeloxifene caused a significant decrease in CartiLaps levels in OVX animals to the same level as found in estrogen-treated or sham-operated animals (FIG. 5). There was no difference between the three levormeloxifene treatment groups. This suggests that it is possible to reduce the dose while maintaining a complete response with respect to cartilage. In contrast, the potential of this compound to inhibit bone resorption has been shown to be less. Only at the highest dose of levormeloxifene was the level of RatLaps (CTX-I) significantly reduced to the same level seen in the sham-operated or estradiol treated groups.

  In conclusion, it has been shown that levormeloxifene is capable of inhibiting cartilage turnover, and therefore, can achieve cartilage erosion in OVX-treated rats to the level seen in normal animals. Furthermore, it has been shown that this SERM exerts a preferential effect on cartilage over bone, and therefore would be extremely useful as a treatment that may protect cartilage in postmenopausal OA.

Levormeloxifen, and levormeloxifen + norethisterone acetate (NETA), are potent inhibitors of cartilage destruction, but do not affect the weight of the uterus of 3 + 1/2 month old rats from which the ovaries have been removed.
To test the protective effects of potential uterus and cartilage, in the OVX model of postmenopausal OA in young animals (3 + 1/2 months), levormeroxifene alone or the gestagen analog compound norethisterone acetate (NETA) The combination was evaluated. Fifty 3 + 1/2 month old female Sprague-Dawley rats were obtained and used in this study. Ten animals were sham operated and the rest (40 animals) were ovariectomized. Animal ovariectomy and handling were performed as described in Example 1. OVX animals were divided into 4 groups of 10 animals, vehicle, levormeroxifene (1.0 mg / kg / day, administered 5 days per week), NETA (1.0 mg / kg / day, 1 week) 5 days administration) or levormeroxifene + NETA (both compounds were administered at 1.0 mg / kg / day, administered 5 days a week) for 5 weeks. All treatments were performed orally in suspension (50% propylene glycol, 0.075 M NaCl). Conducted daily 5 days a week and not administered on weekends. Throughout the duration of the study, animals were weighed at regular intervals and blood / urine samples were collected. Collect urine samples as spots of urine samples by gently urinating the animal's abdomen or by collecting the urine samples obtained by placing the animals in a metabolic measurement cage for 30-60 minutes did.

  Treatment with levormeloxifene showed the same weight development as a sham-operated animal. On the other hand, animal growth was retarded when treated with levormeloxifene + NETA. Treatment with vehicle or NETA increased body weight and the growth rate of the vehicle group was the fastest (Table 1). Compared to vehicle-treated rats, those treated with levormeloxifen, levormeloxifene plus NETA or NETA had no effect on the weight of the uterus of OVX rats (Table 1). Sham-operated rats had an increased uterine weight than OVX rats. This is due to the production of endogenous estrogens. Treatment with levormeloxifene, NETA or levormeloxifene plus NETA resulted in a significant decrease in CartiLaps levels in OVX animals to the same level as found in sham-operated animals (FIG. 6). When comparing CartiLaps and RatLaps data with Examples 1, 2 and 4, this experimental population should be considered younger (3 + 1/2 months of age at the start of the study) and therefore the epiphyseal cartilage plate The presence of type II collagen results in substantial skeletal growth that will contribute to both RatLaps and CartiLaps assessments. Therefore, because of the gradual decline in skeletal growth in this age range, there was an approximately 40% reduction in CartiLaps over the 4-week observation period in the sham-operated group (FIG. 6). . Therefore, it is necessary to consider the increase in bone and cartilage turnover caused by OVX as seen in the vehicle treatment group in conjunction with the sham operation group. Treatment with levormeloxifene or levormeloxifene plus NETA resulted in a significant decrease in CartiLaps and RatLaps levels in OVX animals to the same level as found in sham-operated animals (FIG. 6). Treatment with NETA alone did not significantly reduce CartiLaps levels compared to vehicle-treated animals. However, in the group of animals treated with NETA alone, there was a trend towards decreasing CartiLaps levels.

  In conclusion, the addition of the gestagen analog NETA to levormeloxifene does not diminish the potential cartilage protective effect of SERM. Such potential combination therapies may continue to provide clinical benefits to human subjects that are significantly associated with the estrogenic effects of SERMs on uterine and breast tissues, and are therefore gestagen This may be desirable because the addition of similar compounds provides a well-recognized way to prevent estrogens from stimulating these tissues.

(-)-Cis-3,4-diarylhydroxychroman is a potent inhibitor of cartilage destruction in rats from which the ovaries have been removed.
In order to test the potential cartilage protective effect of this SERM, in the OVX model of postmenopausal OA (see Example 1), the chroman SERM compound (-)-cis-3,4-diarylhydroxychroman (compound # 0781, FIG. 7) was evaluated. Seventy six month old female Sprague-Dawley rats were obtained and used for this study. Ten animals were sham operated and the rest (60 animals) were ovariectomized. Animal ovariectomy and handling were performed as described in Example 1. OVX animals are divided into 5 groups of 12 animals and vehicle, 0.1 mg / kg / day of 17α-ethynylestradiol (Sigma, E-8875, lot number 21K1267) administered 5 days per week, or Treated with either of three doses of (−)-cis-3,4-diarylhydroxychroman (0.2, 1 or 5 mg / kg / day, administered 5 days per week). All treatments were performed orally in suspension (50% propylene glycol, 0.075 M NaCl). Conducted daily 5 days a week and not administered on weekends. Throughout the duration of the study, animals were weighed at regular intervals and blood / urine samples were collected. Collect urine samples as spots of urine samples by gently urinating the animal's abdomen or by collecting the urine samples obtained by placing the animals in a metabolic measurement cage for 30-60 minutes did. RatLaps and CartiLaps measurements (FIGS. 8, 9) demonstrate that this compound has a significant effect on both bone resorption and cartilage turnover. At the highest SERM doses, CartiLaps levels are reduced to those of estrogen or sham-operated animals (left part of FIG. 8), but bone resorption cannot be suppressed to the same extent as estrogen It was. This suggests that this compound has a stronger effect on cartilage than bone. Therefore, this compound may exert a potential cartilage protective function in the treatment of postmenopausal OA.

Delayed development and reduced severity and incidence of inflammatory arthritis induced by (−)-cis-3,4-diarylhydroxychroman in Lewis rats inoculated with type II collagen.
One of the most widely adopted animal models of RA is the Lewis rat collagen-induced arthritis (CIA) animal model. In this model, joint inflammation is induced by inoculation with type II collagen. This inoculation leads to a severe inflammatory response to the joint cartilage, evident as swelling of the foot joint and subsequent destruction of the joint when histological analysis is performed. In this study, we measured the effects of estrogen as well as SERM # 0781 ((−)-cis-3,4-diarylhydroxychroman, FIG. 7) on disease progression and CIA severity. Arthritis was monitored by scoring the swelling and redness of the foot with the naked eye. Cartilage and bone erosion was monitored by quantifying CartiLaps urine levels and RatLaps serum levels. In addition, the weight of the animal's uterus was measured to assess the estrogenicity of the estrogen.

  The study population consisted of 66 female Lewis HanHsd rats (Harlan, The Netherlands). After acclimatization for 1 week, 55 rats were first anesthetized with Hypnorm-Dormicum (1 part of Hypnorm + 1 part of Dormicum + 2 parts of sterile deionized water) Ovariectomy is performed. Rats are given 0.15-0.2 mL / 100 g body weight and standard ovariectomy is performed. Eleven rats are sham operated using the same anesthesia. Rats are inoculated one week after OVX. Each rat is inoculated with 150 μg of bovine type II collagen (Cat # # 2002-1; Chondrex, Redmond, WA, USA) emulsified with Freund's incomplete adjuvant (IFA). 150 μL of the emulsion is injected intradermally at the base of the tail. Every day from day 11 the rat's paw is scored by visual observation of the paw. Each leg was scored on a scale of 0-4 and the scores for the four legs were summed. If the total arthritis score for a given animal reaches 10, or if arthritis for one foot has progressed to a very severe degree, or if the CIA has continued for more than 16 days, more severe arthritis is prominent The animals were killed with ethical considerations because of the pain and discomfort. The rest of the animals were kept for 42 days from OVX before being completed.

  After OVX and sham surgery, 59 of the animals randomly administered to group 6 survived (see Table 3). Ten animals were in the sham-operated group (S), 10 animals were in the group given only vehicle (50% propylene glycol, 0.15 M NaCl) (V), and 11 animals Group (E) was given 0.5 mg / kg / day of estrogen (17α-ethynylestradiol, Sigma, E-8875, lot number 21K1267). Each 10, 8 and 10 animals in the group were then treated with 0.2, 1 and 5 mg / kg / day of SERM # 0781 (A, B and C). The compound is suspended in propylene glycol (Unikem, Copenhagen, Denmark, catalog number 291443) by vortexing and ultrasound. Once a homogenized suspension is obtained, the compound is diluted with the same amount (1: 1) of 0.15 M NaCl. The compound was administered orally by gavage 5 days a week and not on the weekend. The day after OVX, ie before inoculation with type II collagen, treatment with vehicle, estrogen and SERM # 0781 was started at three different doses.

  At the end of the experiment, the weight of the uterus was measured to examine the effect of SERM # 0781 on uterine hypertrophy. Maximum weight was seen in sham-operated animals (S), meaning normal uterine weight. As seen in the vehicle treatment group (V), OVX resulted in more than 50% reduction in uterine weight. Estrogen treatment (E) restores uterine weight to some extent, but is not complete. SERM # 0781 treated rats show a slight increase in uterine weight compared to vehicle treated rats regardless of dose.

  Spleen weights were measured at the end of the CIA experiment. Compared to the sham and vehicle groups, spleen weight was reduced in all SERM # 0781 treated and estrogen groups. The result of statistical analysis of medium dose SERM # 0781 (B) compared to vehicle group (V) is p = 0,014 (n = 5) (FIG. 10). This may suggest the immunosuppressive effect of SERM # 0781, which may be crucial for the progression of CIA.

  In the sham-operated group (S), vehicle group (V), and medium-dose SERM # 0781 group (B), the first signs of arthritis were observed 12 days after inoculation. Consistent with previously published studies, OVX rats were more susceptible to CIA. On day 15 after inoculation, the media group had the highest incidence of CIA and the highest severity of CIA (Table 2).

  High dose SERM # 0781 (C) was fully protected by day 18 (FIG. 11). All three SERM # 0781 treatment groups remained protected and showed a lower arthritic index from day 25 to day 25 of inoculation. However, estrogen-treated rats showed the most severe disease from day 25. It was terminated when the rat score reached 10 (out of 16), or when one of the paws progressed to very severe arthritis, or when CIA continued for more than 16 days. FIG. 12 shows that all three SERM # 0781 groups had improved survival from the inoculation to the first 5 weeks compared to the vehicle group. The estrogen treated group also showed improved survival up to the first 4 weeks. At the end of the experiment, 50% of the low dose SERM # 0781 treated rats were alive, although all other groups had fallen to 20%.

  Taken together, FIGS. 11 and 12 show that SERM # 0781 has a protective effect on the progression of CIA. Interestingly, SERM # 0781 treated rats were less susceptible to CIA than sham-operated groups, suggesting that SERM # 0781 has an additional protective effect rather than simply compensating for estrogen deficiency. .

  CartiLaps analysis of articular cartilage turnover in this young animal population (9 weeks of age at the start of the study) was hampered by significant skeletal growth that occurred during the study period. Skeletal growth is associated with abrupt turnover of the epiphyseal cartilage plate, which is composed of matrix-containing cartilage-like type II collagen, and metabolites from this process will contribute to the measurement of CartiLaps assessment Let's go. Thus, over the 7 week observation period, the overall reduction in animals receiving sham surgery was approximately 50% (FIG. 13A). OVX rats treated with vehicle alone showed increased CartiLaps levels after 3 weeks compared to the sham-operated group. In contrast, estrogen-treated rats showed a decrease in levels (FIG. 13A). SERM # 0781 treatment reduced CartiLaps levels in a dose-dependent manner. The low-dose group behaved similarly to the vehicle group, and as a result of the high-dose treatment, the CartiLaps levels were significantly reduced compared to the vehicle (p = 0,012), comparable to estrogen treatment (p = 0,0021) It became. The great effect of arthritis on the CartiLaps level shown in FIG. 13B explains that the change in the CartiLaps level in FIG. 13A is large. At 3 weeks after OVX (= 2 weeks after type II collagen inoculation), there was a significant difference between treatment groups, but at this point already, the subsequent disease process had an effect on CartiLaps levels. (FIG. 13A).

  At the time of inoculation, the first week of FIG. 13 is one week after OVX. The end point value for rats that remained healthy during the 6 week observation period is 26.5% of the baseline level of CartiLaps. This group (n = 10) also includes rats that developed weak symptoms of CIA (scores of 2 or less) during the last week of observation. Significantly higher endpoint values were found in arthritic rats (p <0,005). Forty-one rats needed to be terminated earlier than planned due to very severe inflammation, i.e., CIA signs (CIA severe / early) over 15 days. Eight rats had moderate disease, i.e., slower progression of CIA and were able to remain in that state until the final end date of week 6 (CIA moderate / late). Both of these CIA groups showed a significant increase in CartiLaps levels at 4 weeks after inoculation (p = 0,0034 and p = 0,014, respectively). Interestingly, the CIA medium / late group already showed a significant level increase at 2 weeks (p = 0,012), which corresponds to 2 weeks before the onset of CIA. This may suggest that elevated CartiLaps levels do not reflect cartilage erosion caused by joint inflammation, but rather reflect changes in cartilage metabolism that occur prior to the onset of arthritis. Such changes in cartilage metabolism may be an early pathological event without clinical symptoms leading to the passage of time and progression of arthritis. Treatment with SERM used in this experiment may be able to suppress this “pre-onset” cartilage turnover and thus reduce the likelihood of progression to subsequent inflammatory arthritis. Thus, SERM compounds may have potential as a prophylactic agent for treating individuals at risk for progression to inflammatory arthritis (RA).

  FIG. 14 shows the absolute value of CartiLaps at the end of the CIA experiment. Different processing groups are shown separately. This reveals that the absolute value of CartiLaps also reflects the state of arthritis in rats. In the OVX OA model, OVX caused an increase in CartiLaps levels (see Examples 1-4). This homeostasis imbalance was repaired within 6 weeks and could be prevented by estrogen treatment, levormeloxifene treatment or SERM # 0781. This period of cartilage metabolism imbalance after OVX may be the reason for the increased susceptibility to cartilage erosion and CIA progression during this period. A further beneficial effect by SERM treatment, as exemplified by the effect of SERM # 0781 in this experiment, may be that they result in a general immunosuppressive effect (see FIG. 10). This effect may then have a significant therapeutic potential for treating inflammatory arthritis.

  In conclusion, the experiments show a significant prophylactic and / or therapeutic effect of SERMs such as the hydroxychroman compound SERM # 0781 in inflammatory arthritis (FIG. 7).

Inhibition of cartilage turnover in humans induced by selective estrogen receptor modulators.
The effect of selective estrogen receptor modulators on cartilage turnover in a population of 235 healthy postmenopausal women aged 49-65 years was evaluated. Cartilage turnover was quantified by measuring the C-telopeptide breakdown products of type II collagen released during cartilage catabolism. For comparison, serum levels of type I collagen C-telopeptide (“CrossLaps”) were measured as a specific marker of bone resorption.

  Participants in the study have been engaged in two different dose determination studies. There, an evaluation of the effects of two different SERMs on the prevention of osteoporosis has been performed. Fasting urine samples were collected periodically throughout the study period and frozen for later study purposes. All participants were more than one year after menopause and the serum estradiol concentration was less than 25 pg / mL. All participants have no clinically apparent acute or chronic disease and have not been given a drug known to affect bone or cartilage metabolism since 6 months prior to enrollment There wasn't. Table 3 reveals the physical demographic data and associated biomarker levels at the participant's baseline. Both studies were conducted according to the Helsinki Declaration II and the practice criteria for standard pharmaceutical clinical trials in Europe.

Design : The raloxifene study included five treatment groups and lasted for five years. One group was given a placebo and three groups were treated with raloxifene, and the oral dose per day was 30, 60 or 150 mg throughout the 5-year study period. In one group, placebo treatment was performed for the first 3 years and then changed to 60 mg raloxifene per day. The study of levormeloxifene included 5 treatment groups and continued for 2 years. One group was given placebo and the four treatment groups were given 1.25 mg, 5 mg, 10 mg and 20 mg of levormeloxifene per day orally, respectively. The treatment group given levormeroxifene for 12 months was followed by 12 months without further treatment. All participants in both studies were given 400-600 mg of calcium supplements per day throughout the study period. In both studies, double-blind processing was performed using a random number table as a method for allocating participants.

Demographic data : Age and years since menopause (YSM) were registered via a baseline question. Height and weight were measured to the nearest 0.1 cm and 0.1 kg, respectively, for participants with light indoor wear and shoes removed. The body mass index (BMI) was calculated as BMI = m / h ² , where m is the patient's body weight in kg and h is the height in meters.

  Urine levels of C-telopeptide fragments of type II collagen were measured by CartiLaps ELISA as described in Example 1.

Serum CrossLaps ELISA : Serum CrossLaps was included for comparison. The serum CrossLaps one-step ELISA is a sensitive indicator of bone resorption and this evaluation was performed as described by the manufacturer (Nrodic Bioscience Diagnostics, Herreb, Denmark). Baseline serum CrossLap measurements were taken at 3, 6 and 12 months after both test treatments.

Statistical analysis : A comparative analysis between study groups at baseline was performed using independent t-test and ANOVA. The correlation between the values of CartiLaps and CrossLaps was examined by Spearman correlation. Results written in letters are mean ± standard deviation. Statistical calculations were performed using SPSS for Windows 10.1 (SPSS, Chicago, Illinois, USA). Differences were considered significant when p <0.05.

  Table 3 provides baseline values for the two study groups divided into treatment groups and their subjects. An independent t-test performed between the raloxifene and levormeloxifene research groups revealed that there was a slight difference between the two groups in terms of age. Therefore, the levormeloxifene group is 1.1 ± 0.5 years (p <0.05) minutes older than the raloxifene group, and the postmenopausal period is correspondingly higher than the raloxifene group. On average, it was 1.9 ± 0.5 years (p <0.05) longer. Serum CrossLaps values reflecting osteoclasts that mediate bone resorption were consistent with such slight differences that the age and YSM of the levormeloxifene group were statistically higher. There were no statistically significant differences between the two study populations with respect to BMI and urine CartiLaps levels reflecting the destruction of cartilage type II collagen. The variance between the parameters listed in Table 1 was determined by independent ANOVA for both study populations. And there was no statistically significant difference between the treatment groups and their controls.

  The upper part of FIG. 15 shows the result of CartiLaps measurement in the raloxifene group from the baseline to the 5-year patrol. In all three doses of raloxifene (30, 60 and 150 mg / day), a significant and sustained reduction of up to about 65% of the baseline of the CartiLaps level was induced, whereas in placebo-treated women There was a slight increase in CartiLaps levels during the 5-year study period. In the treatment group, the CartiLaps marker reached a plateau after 12 months without an apparent dose response. The group that received placebo for the first 3 years, followed by 60 mg raloxifene per day, showed a similar response to established therapies.

  The measurement result of CartiLaps resulting from the study of levormeloxifene is shown in the lower part of FIG. In this study, a 50% reduction in CartiLaps levels was seen 3-6 months after levormeroxifene treatment. Only relatively small and insignificant differences were seen between responses in the four treatment groups during the 12-month treatment intervention period. After discontinuing levormeroxifene treatment, the levels of all CartiLaps in the four treatment groups returned to baseline.

  When the CrossLaps measurements of quantifying bone resorption correlated with their corresponding CartiLaps values, only a slight correlation of r = 0.42 was found. The effects of estrogen and related compounds, such as SERMs, on tissues that respond to estrogen such as the endometrium, bone and breast have been studied in detail. Cartilage is generally not considered a tissue that responds to estrogen, and as already described, there are conflicting reports in the literature on whether estrogen exerts a beneficial protective effect on cartilage. Here we show that two different SERMs induce a significant decrease in urine levels of C-telopeptide fragments of type II collagen, which indicates that these compounds have a significant cartilage protective effect. It has been demonstrated.

  Specific proteolytic destruction of type II collagen is a significant event for cartilage destruction both in vitro and in vivo. Therefore, metabolites derived from this protein are well suited as biochemical markers for cartilage destruction. The evaluation of CartiLaps has been shown in relation to other known biochemical markers that assess cartilage turnover. And in a long-term study of patients with knee OA, CartiLaps measurements were associated with joint space narrowing calculated by radiology and a standardized score for pain and function. Thus, the effect of SERM treatment on CartiLaps levels, as seen in this study, means a direct effect on cartilage turnover throughout the body.

  The study population, which includes two studies, includes postmenopausal women who were not selected according to a specific degree of OA. However, it has already been shown that postmenopausal women experience an increased incidence of OA. If the destruction of cartilage in postmenopausal women can be suppressed, the risk of progression to OA may be reduced accordingly. Thus, postmenopausal women (not selected by showing signs of OA) represent an appropriate group to study treatments that protect cartilage by prevention. Because recorded and approved indications for such treatments include prevention of postmenopausal osteoporosis, postmenopausal women also represent an important target group for SERM treatment. The study population treated with raloxifene was followed for 5 years. In the three groups treated continuously, the decrease in CartiLaps levels appeared completely after 12 months of raloxifene treatment and remained at this level until the end of the study period (Figure 15). This demonstrates not only that the SERM's potential cartilage protective effect is maintained over long-term treatment, but also that the cartilage protective effect of raloxifene appears relatively mildly. Long term treatment is probably needed to provide significant therapeutic benefits. The lack of significant difference between the three doses investigated (30, 60 and 150 mg) suggested that even the lowest dose used in this study provided the greatest cartilage protection effect. When switching to raloxifene for placebo-treated women, it showed a response similar to SERM with a 35% reduction in CartiLaps levels.

  In the study of levormeloxifene, SERMs were given for 12 months and then patients were followed for an additional 12 months. Given 20 mg daily, where the reduction in CartiLaps levels was most noticeable, there was a slight indication of a dose response effect. This reduction appeared completely after 3-6 months of treatment. This demonstrated that the cartilage protective effect appeared earlier when given the tested doses of levormeloxifen compared to raloxifene. Furthermore, the suppression of cartilage destruction markers was also more pronounced compared to the effect seen with raloxifene. When treatment with levormeloxifene was stopped 12 months after treatment, CartiLaps levels returned to baseline values after 12 months. This demonstrated that the potential inhibitory effect of levormeloxifene on cartilage destruction is reversible.

  Both raloxifene and levormeloxifene inhibit bone resorption, as measured by biomarkers, and in the case of raloxifene, as can be seen by the long-term decrease in the incidence of long-term fractures. The effect of SERMs on bone resorption as assessed by CrossLaps ELISA was comparable to the effect on cartilage destruction as we demonstrated here by the CartiLaps assessment method.

  Although the response to SERM as measured by the CartiLaps and CrossLaps evaluations appears to be similar, the two evaluations clearly represent different processes. First, the correlation between the CartiLaps measurement and the CrossLaps measurement was weak (r = 0.42). More importantly, some differences can be noted in the effect of SERMs on cartilage and bone turnover. For raloxifene, all three doses tested had the greatest effect on cartilage destruction, whereas the effect on bone resorption showed signs of a dose response. Furthermore, although raloxifene required 12 months of treatment to maximize cartilage destruction, bone resorption markers were completely suppressed after only 3-6 months. For levormeloxifene, there was a trend for a dose-response effect in CartiLaps, whereas levormeloxifene reduced CrossLaps to the same level at all doses. Thus, bone and cartilage responsiveness to levormeloxifene is quantitatively different, indicating that different doses are required to have the greatest effect on bone and cartilage turnover. These observations demonstrate that the kinetics of the effects exerted by these SERMs on bone and cartilage are different.

  In conclusion, the results demonstrate that treatment with compounds such as estrogens or estrogen-like substances such as SERMs, which function like estrogen agonists, can induce significant suppression of cartilage catabolism. Since cartilage destruction is one of the most prominent features of OA, inhibiting or suppressing this process may lead to reduced prevalence or reduced severity of pathological processes leading to OA. Thus, SERMs represent a novel therapeutic option component for treating or preventing OA.

References:

Cartilage erosion in four different condyles when 5 month old (upper) and 7 month old (lower) female rats were raised for 9 weeks from OVX or sham surgery treatment. Express erosion as a percentage of the total surface of the cartilage. For the two groups (ovx-dark green, sham surgery-light green), erosion is expressed as mean erosion + standard error value. Each of the four condyles was scored. Medial tibia (Medium T), medial femur (Medium F), lateral tibia (Lateral T), and lateral femur (Lateral F). The final, average of all four regions is shown (total). Cartilage and bone turnover in OVX and sham-operated rats. Bone resorption was determined by assessing cartilage turnover using CTX-II markers (part A, part B) and measuring RatLaps (CTX-I) (part C, part D). The left part (A, C) is a measurement from a rat that was 5 months old and followed for 9 weeks at the start. The right part (B, D) is a measurement from a rat that was 7 months old and followed for 9 weeks at the start. The X axis shows the number of weeks after OVX. Correlation between biomarker CartiLaps expressed in% of baseline (4 weeks after surgery) and histological scoring of erosion (n = 16) for experiments on 7 month old animals. A linear regression yielded a correlation coefficient of R ² = 0.27 and a significant p-value of 0.039. Percentage of erosion (at 4 weeks) in animals with low or high values of CartiLaps in the 7 month old population. Effect of levormeloxifene on cartilage turnover (left) and bone resorption (right) in OVX rats. Results are expressed as the mean% of baseline within the group and error bars represent standard error (SEM). Effect of levormeloxifene, levormeloxifene + NETA, and NETA on bone resorption (left) and cartilage destruction (right) in ovariectomized rats. Results are expressed as the mean% of baseline within the group and error bars represent standard error (SEM). Chemical structure of (−)-cis-7-hydroxy-3-phenyl-4- [4- [2- (pyrrolidin-1-yl) ethoxy] phenyl] chroman of SERM # 0781. Three doses of (−)-cis-3,4-diarylhydroxychroman (SERM # 0781), and CartiLaps (left) and RatLaps (right) levels in 6-month-old OVX rats treated with vehicle and estradiol . Measurement of area under the curve (AUC) of the CartiLaps data of FIG. Student's t-test is performed to test for significant differences with the vehicle treatment group. ^* ) P>0.05; ^** ) p <0.01; NS) not significant. Spleen weight at the end of the CIA-2 experiment. A) low dose SERM # 0781, B) medium dose SERM # 0781, C) high dose SERM # 0781, E) estrogen, S) sham surgery, V) vehicle. The value is expressed as an average value with standard error. A) low dose SERM # 0781, B) medium dose SERM # 0781, C) high dose SERM # 0781, E) estrogen, S) sham surgery, V) treated with vehicle or treated with SERM # 0781 Cumulative incidence of arthritis in CIA animals that did not. Values are expressed as mean values within the group. The arthritic index is the average of the arthritic clinical scores of all rats in the group. The X axis indicates the number of days after inoculation with type II collagen (performed after 1 week of OVX). A) low dose SERM # 0781, B) medium dose SERM # 0781, C) high dose SERM # 0781, E) estrogen, S) sham surgery, V) treated with vehicle or treated with SERM # 0781 Mean survival time in CIA animals that were not. Values are given as percent of surviving rats within each group. The X axis indicates the number of days after inoculation with type II collagen (performed after 1 week of OVX). A) low dose SERM # 0781, B) medium dose SERM # 0781, C) high dose SERM # 0781, E) estrogen, S) sham surgery, V) treated with vehicle or treated with SERM # 0781 CartiLaps levels in CIA animals that were not present. Values are given as the mean value with standard error within each group for the% baseline calculated for each animal, with its own baseline value as the standard. In the right part (B), the arthritis score of animals with mild or very mild disease (score of 2 or less), the arthritis score of animals with late onset of arthritis (after 25 days), and disease during the study period The total animal population is divided according to the arthritic score of animals with an early onset or severely severe arthritis. The average response, calculated as a percentage of the individual baseline, is plotted again with error bars representing SEM. Week 0 on the X-axis indicates the time point of OVX. Difference in CartiLaps levels divided according to the presence or absence of clinical arthritis in CIA animals treated or not with SERM # 0781. The display of the group is as follows. A) low dose SERM # 0781, B) medium dose SERM # 0781, C) high dose SERM # 0781, E) estrogen, S) sham surgery, V) vehicle. Values are given as mean creatinine values corrected for values with standard deviation for each group. Changes in urinary CartiLaps, a cartilage disruption marker that measures the product of C-telopeptide of type II collagen disrupted during treatment with raloxifene (top) and levormeloxifene (bottom). The design and treatment groups are described in the method. Error bars indicate standard error.

Claims (39)

  Droloxifene, levormeloxifene, chroman derivatives, nafoxidine, bazedoxifene, miproxyfen, arzoxifene, lasofoxifene, MDL-103323, EM-800, fulvestrant, ICI 183,780, ICI 164,384 , GW 5638, 19-nortestosterone derivative, and a pharmaceutically acceptable ester, ether and salt thereof, an effective amount of a selective estrogen receptor modulator (SERM), comprising administering to the subject Methods for preventing, treating or alleviating symptoms including sexual cartilage destruction.   2. The method of claim 1, wherein the SERM comprises levormeloxifene, a chroman derivative or a 19-nortestosterone derivative, or a pharmaceutically acceptable salt thereof. The SERM is represented by the formula 2:

The method according to claim 1, comprising a core structure represented by   4. A method according to claim 1 or claim 3, wherein the SERM is a chroman derivative. Said selective estrogen receptor modulator is of formula I

(Where
R ¹ is H, SO ₂ NR ₂ ⁴ , SO ₂ NHR ⁴ , Cl, CH ₃ or benzyl;
R ² is independently selected from the group consisting of OH, halogen, nitro, cyano, SH, SR ⁴ , trihalo-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy and phenyl. Phenyl optionally substituted with 1 to 5 substituents,
R ³ is phenyl substituted with --X-(CH ₂ ) _n --Y, where
X is a valence bond, O or S,
n is an integer ranging from 1 to 12,
Y is H, halogen, OH, OR ⁴ , NHR ⁴ , NR ₂ ⁴ , NHCOR ⁴ , NHSO ₂ R ⁴ , CONHR ⁴ , CONR ⁴ , COOH, COOR ⁴ , SO ₂ R ⁴ , SOR ⁴ , SONR ⁴ , SONR ₂ ⁴ , a pyrrolidinyl ring, independent of the group consisting of H, OH, halogen, nitro, cyano, SH, SR ⁴ , trihalo-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl and C ₁ -C ₆ alkoxy Optionally substituted with 1 to 3 substituents selected by
R ⁴ is C ₁ -C ₆ alkyl. )
5. The method of claim 4, comprising chroman derivatives of and optical and geometric isomers, pharmaceutically acceptable esters, ethers and salts thereof. R ² and R ³ are arranged in the transformer, The method of claim 5.   Use of a chroman compound which is (−)-3,4-trans-7-methoxy-2,2-dimethyl-3-phenyl-4 {4- [2- (pyrrolidin-1-yl) ethoxy] phenyl} chroman. The method of claim 6 comprising. Where
R ¹ is H, SO ₂ NR ₂ ⁴ , CH ₃ or SO ₂ NHR ⁴ ;
R ² and R ³ are arranged in cis,
R ² is independently selected from the group consisting of OH, halogen, nitro, cyano, SH, SR ₄ , trihalo-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl and C ₁ -C ₆ alkoxy. Phenyl optionally substituted with ~ 3 substituents,
R ³ is phenyl substituted with --X-(CH ₂ ) _n --Y, where
X is a valence bond, O or S,
n is an integer ranging from 1 to 12,
Y represents H, OH, OR ⁴ , NHR ⁴ , NR ₂ ⁴ , NHCOR ⁴ , NHSO ₂ R ⁴ , CONH ⁴ , CONR ₂ ⁴ , COOH, COOR ⁴ , SO ₂ R ⁴ , SOR ⁴ , SONHR ⁴ , SONR ₂ ⁴ , A pyrrolidinyl ring, independently selected from the group consisting of H, OH, halogen, nitro, cyano, SH, SR ⁴ , trihalo-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl and C ₁ -C ₆ alkoxy Optionally substituted with 1 to 3 substituents,
R ⁴ is C ₁ -C ₆ alkyl;
And the optical isomers and geometric isomers, pharmaceutically acceptable esters, ethers and salts thereof. A formula wherein R ¹ , R ² and R ³ are as defined above:

Or

9. The method of claim 8, comprising the use of a compound of formula Ia or formula Ib having: A formula wherein R ¹ is as defined above and m is an integer from 0 to 10:

9. The method of claim 8, comprising the use of a compound of: A formula wherein R ¹ is as defined above and R ⁶ represents one or more of the following substituents: methoxy, hydroxy, trifluoromethyl, fluoro and chloro:

9. The method of claim 8, comprising the use of a compound of: (−)-Cis- {3-phenyl-4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-chroman-7-ol;
(−)-Cis- {3- (4-trifluoromethylphenyl) -4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-7-methoxychroman;
(− / +)-Cis- {3- (3-methoxyphenyl) -4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-7-methoxychroman;
(− / +)-Cis- {3- (3-hydroxyphenyl) -4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-chroman-7-ol;
(− / +)-Cis- {3- (4-trifluoromethylphenyl) -4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-chroman-7-ol; or (+ ,-)-(Cis-3- (4-methylphenyl) 4- (4- (2-pyrrolidinoethoxy) phenyl) -chroman-7-yl) N, N-dimethylsulfamic acid ester;
(−)-Cis-7-methoxy-3-phenyl-4- (4- (2-pyrrolidinoethoxy) -phenyl) chroman;
(+/-)-cis-7-methoxy-3- (3-methoxyphenyl) -4- (4- (2-pyrrolidinoethoxy) -phenyl) chroman;
(+/-)-cis-7-methoxy-3- (4-trifluoromethylphenyl) -4- (4- (2-pyrrolidinoethoxy) -phenyl) chroman; and (-)-cis-7-methoxy 12. A process according to claim 11 comprising the use of a chroman compound which is -3- (4-trifluoromethylphenyl) -4- (4- (2-pyrrolidinoethoxy) -phenyl) chroman.   13. Use of a SERM according to any one of claims 1 to 12 for the preparation of a medicament for use in preventing, treating or alleviating symptoms including progressive cartilage destruction. 1) together with a selective estrogen receptor modulator (SERM) or a pharmaceutically acceptable salt thereof,
2) A method for preventing, treating or alleviating symptoms including progressive cartilage destruction, comprising administering to a subject an effective amount of a combination of progestin or a pharmaceutically acceptable salt thereof.   The selective estrogen receptor modulator (SERM) is raloxifene, droloxifene, tamoxifen, 4-hydroxy-tamoxifen, 4′-iodotamoxifen, toremifene, (deaminohydroxy) toremifene, clomiphene, levormeloxifene, chroman derivative , Coumarin derivatives, idoxifene, nafoxidine, basedoxifene, miproxyfen, arzoxifene, lasofoxifene, MDL-103323, EM-800, full restaurant, ICI 183,780, ICI 164,384, GW 5638, diethylstil Bestrol, genistein, nafoxidine, nitromiphene citrate, moxesterol, diphenol hydrochrysene, erythro-MEA, arenolic acid, Selected from the group consisting of clophenyl, chlorotrianicene, ethamoxytrifertol, dehydroepiandrosterone, triparanol 19-norprogesterone derivatives, 19-nortestosterone derivatives and pharmaceutically acceptable esters, ethers and salts; The progestin is norethindrone, ethinodiol, desogestrel, levonorgestrel, norgestrel, norgestimate, medroxyprogesterone, danazol, linoestrenol, didrogesterone, chlormadinone, promegesterone, gestrinone, algestone acetophenide, allylestrenol acetate, cyproterone acetate , Demegestone, Guestden, Osaterone, Hydroxyprogesterone hexanoate, Medrogestone, Megestro , Nomegestrol, ethynylnortestosterone, norpregnenolone, NSC-9564, norethinodrel, dexnorgestrel, guestden, progesterone, chlormadinone acetate, drospirenone (dihydrospirorenone), or 3-ketodesogestrel and optical isomers 15. The method of claim 14, wherein the method is selected from the group consisting of: and geometric isomers, pharmaceutically acceptable esters and salts thereof.   The SERM is raloxifene, droloxifene, tamoxifen, levormeloxifene, chroman derivative, coumarin derivative, idoxyphene, nafoxidine, bazedoxifene, toremifene, (deaminohydroxy) toremifene, miproxyfen, arzoxifene, lasofoxifene , MDL-103323, EM-800, Fulvestrant ICI 183,780, GW 5638 and 19-nortestosterone derivatives or pharmaceutically acceptable salts thereof. The SERM is shown in FIG.

The method according to claim 15, comprising a core structure represented by   17. A method according to claim 15 or claim 16, wherein the SERM is a chroman derivative. Said selective estrogen receptor modulator is of formula I

(Where
R ¹ is H, SO ₂ NR ₂ ⁴ , SO ₂ NHR ⁴ , Cl, CH ₃ or benzyl;
R ² is independently selected from the group consisting of OH, halogen, nitro, cyano, SH, SR ⁴ , trihalo-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy and phenyl. Or phenyl optionally substituted with 1 to 5 substituents.
R ³ is phenyl substituted with --X-(CH ₂ ) _n --Y, where
X is a valence bond, O or S,
n is an integer ranging from 1 to 12,
Y is H, halogen, OH, OR ⁴ , NHR ⁴ , NR ₂ ⁴ , NHCOR ⁴ , NHSO ₂ R ⁴ , CONHR ⁴ , CONR ⁴ , COOH, COOR ⁴ , SO ₂ R ⁴ , SOR ⁴ , SONR ⁴ , SONR ₂ ⁴ , a pyrrolidinyl ring, independent of the group consisting of H, OH, halogen, nitro, cyano, SH, SR ⁴ , trihalo-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl and C ₁ -C ₆ alkoxy Optionally substituted with 1 to 3 substituents selected by
R ⁴ is C ₁ -C ₆ alkyl. )
19. The method of claim 18 comprising the chroman derivatives of and optical and geometric isomers, pharmaceutically acceptable esters, ethers and salts thereof. R ² and R ³ are arranged in the transformer, The method of claim 19.   Use of a chroman compound which is (−)-3,4-trans-7-methoxy-2,2-dimethyl-3-phenyl-4 {4- [2- (pyrrolidin-1-yl) ethoxy] phenyl} chroman. 21. The method of claim 20, comprising. 20. A method according to claim 19, wherein
R ¹ is H, SO ₂ NR ₂ ⁴ , CH ₃ or SO ₂ NHR ⁴ ;
R ² and R ³ are arranged in cis,
R ² is independently selected from the group consisting of OH, halogen, nitro, cyano, SH, SR ₄ , trihalo-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl and C ₁ -C ₆ alkoxy. Phenyl optionally substituted with one substituent,
R ³ is phenyl substituted with --X-(CH ₂ ) _n --Y, where
X is a valence bond, O or S,
n is an integer ranging from 1 to 12,
Y represents H, OH, OR ⁴ , NHR ⁴ , NR ₂ ⁴ , NHCOR ⁴ , NHSO ₂ R ⁴ , CONH ⁴ , CONR ₂ ⁴ , COOH, COOR ⁴ , SO ₂ R ⁴ , SOR ⁴ , SONHR ⁴ , SONR ₂ ⁴ , A pyrrolidinyl ring, independently selected from the group consisting of H, OH, halogen, nitro, cyano, SH, SR ⁴ , trihalo-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl and C ₁ -C ₆ alkoxy Optionally substituted with 1 to 3 substituents,
R ⁴ is C ₁ -C ₆ alkyl;
And optical and geometric isomers, pharmaceutically acceptable esters, ethers and salts thereof. A formula wherein R ¹ , R ² and R ³ are as defined above:

Or

23. The method of claim 22, comprising the use of a compound of formula Ia or formula Ib having: A formula wherein R ¹ is as defined above and m is an integer from 0 to 10:

24. The method of claim 22, comprising the use of a compound of: A formula wherein R ¹ is as defined above and R ⁶ represents one or more of the following substituents: methoxy, hydroxy, trifluoromethyl, fluoro and chloro:

24. The method of claim 22, comprising the use of a compound of: (−)-Cis- {3-phenyl-4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-chroman-7-ol;
(−)-Cis- {3- (4-trifluoromethylphenyl) -4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-7-methoxychroman;
(− / +)-Cis- {3- (3-methoxyphenyl) -4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-7-methoxychroman;
(− / +)-Cis- {3- (3-hydroxyphenyl) -4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-chroman-7-ol;
(− / +)-Cis- {3- (4-trifluoromethylphenyl) -4- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl]}-chroman-7-ol; or (+ ,-)-(Cis-3- (4-methylphenyl) 4- (4- (2-pyrrolidinoethoxy) phenyl) -chroman-7-yl) N, N-dimethylsulfamic acid ester;
(−)-Cis-7-methoxy-3-phenyl-4- (4- (2-pyrrolidinoethoxy) -phenyl) chroman;
(+/-)-cis-7-methoxy-3- (3-methoxyphenyl) -4- (4- (2-pyrrolidinoethoxy) -phenyl) chroman;
(+/-)-cis-7-methoxy-3- (4-trifluoromethylphenyl) -4- (4- (2-pyrrolidinoethoxy) -phenyl) chroman; and (-)-cis-7-methoxy 26. The method of claim 25 comprising the use of a chroman compound that is -3- (4-trifluoromethylphenyl) -4- (4- (2-pyrrolidinoethoxy) -phenyl) chroman.   27. The progestin is any one of claims 14 to 26, wherein the progestin is progesterone, norethindrone acetate, ethinodiol, desogestrel, levonorgestrel, norgestrel, norgestimate, medroxyprogesterone or an ester or a pharmaceutically acceptable salt thereof. Method.   28. The method of claim 27, wherein the progestin is norethindrone or norethindrone acetate or an ester or a pharmaceutically acceptable salt thereof.   A factor that inhibits the production of estrogen, wherein the composition comprises a SERM that is a 3,4 chroman or coumarin derivative or a pharmaceutically acceptable salt thereof, and a progestin or a pharmaceutically acceptable salt thereof; A pharmaceutical composition for use in therapeutic or prophylactic treatment of a disease, comprising an effective amount of a combination of and a pharmaceutical carrier or diluent.   28. A method comprising administering an effective amount of the pharmaceutical composition of claim 27 to a mammal in need thereof, breast cancer, testicular cancer, osteopenia, ie bone loss leading to osteoporosis, endometriosis, heart disease, high cholesterol Hazards related to exposure to chemicals in the environment, blood pressure, enlarged prostate, prostate cancer, obesity, hot flashes, skin effects, mood swings, memory loss, urinary incontinence, hair loss, cataracts, natural hormone disorders Therapeutic or prophylactic treatment of symptoms such as reproductive toxicity.   Due to progressive cartilage destruction, the composition comprises a 3,4 chroman or coumarin derivative or a pharmaceutically acceptable salt thereof SERM and a progestin or a pharmaceutically acceptable salt thereof A pharmaceutical composition for use in the therapeutic or prophylactic treatment of a disease, comprising an effective amount of a salt and a pharmaceutical carrier or diluent.   30. The pharmaceutical composition according to claim 27 or 29, wherein the progestin is norethindrone or norethindrone acetate or a pharmaceutically acceptable salt thereof.   A dosage form suitable for administering 0.01 to about 100 mg of an effective amount of the chroman or coumarin derivative or pharmaceutically acceptable salt thereof per person per day, as claimed in claim 29 or claim 31. The pharmaceutical composition as described.   35. The pharmaceutical composition according to claim 32, in a dosage form suitable for administering 0.01 to about 100 mg as an effective amount of norethindrone or norethindrone acetate or a pharmaceutically acceptable salt thereof per person per day. .   35. A pharmaceutical composition according to any one of claims 29 to 34, which is in the form of an oral unit dose or a parenteral unit dose.   A method of treating osteoarthritis comprising administering to a mammal in need thereof a therapeutically effective amount of a combination of a compound of formula (I) and a progestin.   A method of treating rheumatoid arthritis comprising administering to a mammal in need thereof a therapeutically effective amount of a combination of a compound of formula (I) and a progestin.   36. A method of treating osteoarthritis or rheumatoid arthritis according to claim 34 or claim 35, wherein the mammal is a human. 36. Use of a cartilage specific biochemical marker for titrating a dose according to claim 34 or claim 35.

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