WO 03/061567
`
`PCT/US03/01120
`
`reducing
`
`
`
`The invention also
`of
`a method
`encompasses
`activation
`of
`the
`preventing the
`
`
`
`SIPl/Edgl in a mammalian patient receptor need in
`thereof comprising administering
`to
`compound
`said
`patient
`a
`the S IPi/Edgl
`receptor
`in
`an
`amount
`effective
`for
`reducing
`activation of S1P1/EDG1
`receptor,
`wherein
`said
`
`compound selectivity for
`possesses
`the SIPl/Edgl
`receptor
`
`over SIPRs/EdgS the
`
`receptor
`of at
`
`least fold as measured 20
`
`by the ratio of EC50 for
`the S IPi/Edgl
`receptor
`
`to EC50 for the SIPs/EdgS the
`
`
`receptor as evaluated in the SSs-GTPyS binding
`assay
`and
`wherein
`possesses an EC50
`for
`
`binding to the SIPl/Edgl
`receptor
`of
`100
`nM
`35S-GTPYS binding
`evaluated by the
`assay,
`
`said
`or
`
`5
`
`10
`
`with the proviso
`
`that
`
`the
`
`
`
`compound fall within formula A: does not
`
`
`
`
`
`R1a
`
`CH2R3
`I
`0=p— X-CH2—C-CH2CH2
`N(R2)2
`1b R
`
`A
`
`Y-R4
`
`15
`
`or a pharmaceutically
`
`
`
`acceptable or hydrate thereof, wherein: salt
`
`
`
`X is O, S, NRl or
`
`(CH2)l-2>
`
`
`
`optionally substituted with 1-4 halo groups;
`
`Rl is H, Ci-4alkyl or haloCi-4
`
`alkyl;
`
`20
`
`the alkyl and alkyl portions being optionally
`R l a is H, OH, Ci_4alkyl, or OCi_4 alkyl,
`substituted with 1-3
`halo
`groups;
`
`-177-
`
`SUN - IPR2017-01929, Ex. 1014, p. 651 of 970
`
`

`

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`PCT/US03/01120
`
`Rib
`
`represents H, OH, Ci_4 alkyl or haIoCi-4 alkyl;
`
`each R2 is independently selected from the group consisting of: H, Ci-4 alkyl and
`
`haloCi-4 alkyl.
`
`R3 is H, OH, halo, Ci_4alkyl, OCi-4alkyl, 0-haloCi-4alkyl
`
`or
`
`hydroxyCi-4aIkyl,
`
`Y is selected from
`and S, and
`
`the group consisting of: -CH2-, -C(O)-, -CH(OH)-, -C(=NOH)-, O
`
`R4 is selected from the group
`
`
`
`consisting of: C4_i4alkyl and C4-I4alkenyl.
`
`infiltration
`The invention also encompasses a method of inhibiting an
`
`need
`thereof
`of a lymphocyte
`into a respiratory tissue in a mammalian patient
`in
`preventing
`promoting a
`sequestration
`of
`
`the in a lymph node lymphocyte
`
`thereby
`respiratory
`release of a pro-inflammatory
`mediator
`in
`the comprising
`
`of
`the
`administering
`to
`said
`patient a compound which
`
`is
`agonist
`an
`receptor in an amount
`
`effective for modulating airway
`function,
`wherein
`said
`compound possesses
`a
`
`selectivity for the SIPl/Edgl
`receptor
`
`
`the over S1PR3/Edg3
`ratio of EC50 for the SIPl/Edgl
`20
`receptor of at
`
`least fold as measured by
`the
`receptor to the EC50 for the SIPs/EdgS
`the 35S-GTPYS
`receptor
`
`as evaluated
`in
`EC50
`
`
`said binding assay and wherein compound possesses
`an
`for binding to the
`100 nM
`35S-GTPYS binding assay,
`SIPl/Edgl
`receptor
`of
`or less as evaluated
`by
`the
`
`with the proviso
`
`that
`
`the
`
`
`
`compound does not fall within
`
`formula
`
`A:
`
`by
`
`tissue
`SIPi/Edgl
`
`5
`
`10
`
`15
`
`20
`
`25
`
`-178-
`
`SUN - IPR2017-01929, Ex. 1014, p. 652 of 970
`
`

`

`WO 03/061567
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`PCT/US03/01120
`
`Rla
`
`CH2R3
`O=P—X-CH2—C-CH2CH2^^.
`'
`N(R2)2
`
`R 1 b
`
`Y-R4
`
`A
`
`or a pharmaceutic ally acceptable
`
`salt
`
`or
`
`thereof, wherein:
`hydrate
`
`X is O, S, NRl or (CH2)l-2> optionally
`
`substituted
`
`with
`
`1-4
`
`halo
`
`Rl is H, Ci-4alkyl
`
`or
`
`haloCi-4
`alkyl;
`
`is H, OH, Ci-4alkyl, or OCi_4 alkyl, the alkyl and alkyl portions
`Rla
`groups;
`halo
`substituted with 1-3
`
`being
`
`optionally
`
`R i b represents H, OH, Ci_4 alkyl or
`
`haloCi-4
`alkyl;
`
`independently
`each R2 is
`haloCi-4 alkyl.
`
`H, Ci_4 alkyl and
`of:
`from the group consisting
`selected
`
`R3 is H, OH, halo, Ci_4alkyl, OCi-4alkyl, 0-haloCi-4alkyl
`
`hydroxyCi-4alkyl,
`or
`
`Y is selected from the group
`and S, and
`
`-CH2-, -C(O)-,
`of:
`consisting
`
`-C(=NOH)-, O
`-CH(OH)-,
`
`R4 is selected from the group consisting of: C4-l4alkyl and C4-l4alkenyl.
`
`5
`
`10
`
`15
`
`20
`
`-179-
`
`SUN - IPR2017-01929, Ex. 1014, p. 653 of 970
`
`

`

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`PCT/US03/01120
`
`WHAT IS CLAIMED IS:
`
`in a
`immunoregulatory abnormality
`A method of treating an
`
`comprising administering
`to said patient
`in
`need of such treatment
`
`5 mammalian patient
`
`is an agonist of the
`SIPi/Edgl
`receptor in an amount effective for
`
`a compound which
`treating said immunoregulatory
`abnormality,
`wherein
`said
`compound
`of
`selectivity for the SIPl/Edgl
`receptor
`
`over S1PR3/Edg3 the
`
`receptor
`at
`least
`
`fold as measured by
`the
`ratio of EC50 for
`
`the SIPi/Edgl
`
`receptor the EC50 for the to
`receptor as evaluated in the SSs-GTPyS binding assay and wherein
`10 SlP3/Edg3
`said
`compound possesses
`an
`EC50 for binding to the SIPi/Edgl
`
`receptor
`of
`100 or
`
`the SSs-GTPyS
`less as evaluated by
`
`binding assay.
`
`nM
`
`possesses
`20
`
`with the proviso
`
`
`
`that the compound does not fall within
`
`formula
`
`A:
`
`1a R
`
`CH2R3
`0=P—X-CH2—C-CHgCHg-^^^
`N(R2)2
`1b R
`
`4
`Y-R
`
`15
`
`A
`
`or a pharmaceutically
`
`
`
`acceptable salt or hydrate thereof, wherein:
`
`
`
`X is O, S, NRl or (CH2)l-2> optionally substituted with 1-4
`
`halo
`
`groups;
`
`20 Rl is H, Ci-4alkyl or haloCi-4 alkyl;
`
`
`
`R l a is H, OH, Ci-4alkyl, or OCi-4 alkyl, the alkyl and alkyl portions being optionally
`
`substituted with 1-3
`
`halo
`
`groups;
`
`-180-
`
`SUN - IPR2017-01929, Ex. 1014, p. 654 of 970
`
`

`

`WO 03/061567
`
`PCT/U S03/01120
`
`Rib represents H, OH, Ci_4 alkyl or haloCi_4
`
`alkyl;
`
`each R2 is independently selected from the group consisting of: H, Ci-4 alkyl and
`
`haloCi_4 alkyl.
`
`R3 is H, OH, halo, Ci-4alkyl, OCi-4alkyl, 0-haloCi-4alkyl
`
`or
`
`hydroxyCi-4alkyl,
`
`Y is selected from the group consisting of: -CH2-, -C(0)-,
`and S, and
`
`-CH(OH)-, -C(=NOH)-, O
`
`R4 is selected from the group consisting of: C4_i4alkyl and C4_i4alkenyl.
`
`has
`The method according to Claim 1 wherein the compound
`
`100
`the SIPi/Edgl
`receptor
`
`over the SlPg/EdgS receptor of at
`least
`selectivity for
`EC50
`EC50 for the
`fold as measured by
`the
`
`ratio for the S IPi/Edgl of
`
`receptor
`to
`the
`35S-GTPYS binding
`SIPs/EdgS
`receptor
`as
`evaluated
`in
`the
`assay.
`
`The method according
`3.
`has
`compound
`2 Claim wherein the
`
`
`200
`
`
`the receptor of at SIPs/EdgS
`least
`the SIPi/Edgl
`receptor
`over
`selectivity for
`EC50
`to the EC50 for the
`
`fold as measured by
`the
`ratio
`of the SIPi/Edgl for
`
`receptor
`SIPs/EdgS
`receptor
`as evaluated in
`
`the SSs-GTPyS binding
`assay.
`
`to
`
`a
`
`a
`
`Claim 3 wherein the
`
`to
`The method according
`4.
`
`the SIPi/Edgl
`receptor
`
`the over SIPs/EdgS
`receptor
`selectivity for
`ratio of EC50 for the SIPi/Edgl
`fold as measured by
`the
`receptor
`in the 35S-GTPYS binding
`SlP3/Edg3
`receptor
`as
`evaluated
`assay.
`
`compound a
`has
`
`500
`of at
`least
`EC50 for the
`to the
`
`5.
`selectivity for
`
`to
`The method according
`the SIPi/Edgl
`receptor
`over the
`
`compound
`4 Claim wherein the
`
`
`
`SlP3/Edg3 receptor of at
`least
`
`has
`2000
`
`a
`
`5
`
`10
`
`15
`
`20
`
`25
`
`-181-
`
`SUN - IPR2017-01929, Ex. 1014, p. 655 of 970
`
`

`

`WO 03/061567
`
`PCT/US03/01120
`
`fold as measured by the
`receptor as
`SlP3/Edg3
`
`
`of EC50 SIPi/Edgl for
`
`
`
`ratio
`the receptor the EC50 for the to
`in the 35S-GTPYS binding assay.
`evaluated
`
`
`
`a
`
`patient
`said
`effective
`compound
`100
`
`in
`immunoregulatory abnormality
`
`an
`A method of treating
`6.
`to
`in
`need
`
`such of treatment comprising
`
`administering
`5 mammalian patient
`amount
`is
`an
`
`agonist SIPi/Edgl
`of
`
`the receptor
`in
`an
`a compound which
`
`treating said
`immunoregulatory
`abnormality,
`wherein
`said possesses a
`least
`
`selectivity for
`the SIPi/Edgl
`receptor
`
`over SlP3/Edg3 the
`receptor
`of at
`EC50
`fold as measured
`by the ratio of EC50
`
`for the S IPi/Edgl
`
`receptor
`to
`
`the for the
`S lP3/Edg3
`
`the 35S-GTPYS binding assay
`receptor
`
`as evaluated
`in
`and wherein said
`10 of or less
`compound possesses
`
`an EC50 for binding
`to the SIPi/Edgl
`receptor
`
`as evaluated by the 35s-GTPyS
`binding
`assay.
`
`10
`
`
`
`nM
`
`15
`
`20
`
`25
`
`30
`
`the compound
`
`The method according Claim 6 wherein to
`
`7.
`
`
`possesses an EC50 for binding to the SIPi/Edgl
`
`receptor of 1 nM or less as evaluated
`by the SSs-GTPyS binding
`assay.
`
`to Claim 6 wherein the
`The method according
`8.
`the SIPi/Edgl
`receptor
`
`over SIPs/EdgS the
`
`receptor
`for
`selectivity
`fold as measured
`by
`the
`ratio
`
`of EC50 SIPl/Edgl
`
`for receptor
`35S-GTPYS binding
`SlP3/Edg3
`
`as receptor evaluated
`
`in
`
`the
`
`has
`compound
`least
`of at
`
`to
`the for the the
`assay.
`
`
`
`a
`200
`EC50
`
`The method according to Claim 8 wherein the
`
`selectivity for the SIPi/Edgl
`
`
`over receptor the SlP3/Edg3
`receptor
`ratio of EC50 for
`fold as measured by
`the
`the SIPi/Edgl
`receptor
`SlP3/Edg3 receptor as evaluated
`in
`the
`
`SSs-GTPyS binding assay.
`
`has
`compound
`of at
`least
`to
`the for the
`
`
`a
`500
`EC50
`
`wherein 9 compound has a
`
`Claim
`to
`The method according
`
`10.
`the SIPi/Edgl
`receptor
`
`over the SlP3/Edg3 receptor of at
`least
`1000
`selectivity for
`
`fold as measured by the ratio of EC50 for the SIPl/Edgl
`
`receptor the EC50 for the to
`
`S1PR3/Edg3
`receptor as evaluated in the SSs-GTPyS binding
`assay.
`
`the
`
`-182-
`
`SUN - IPR2017-01929, Ex. 1014, p. 656 of 970
`
`

`

`WO 03/061567
`
`PCT/US03/01120
`
`compound
`10 wherein the
`Claim
`to
`The method according
`11.
`SIPRs/EdgS
`the
`over
`receptor
`the SIPi/Edgl
`for
`a selectivity
`EC50
`of EC50 for
`receptor
`the SIPi/Edgl
`ratio
`the
`by
`as measured
`2000 fold
`35S-GTPYS binding
`assay.
`the
`in
`evaluated
`as
`receptor
`for the SIPs/EdgS
`
`has
`receptor
`to the
`
`1 wherein the
`Claim
`to
`The method according
`12.
`inflammatory disease
`chronic
`or
`abnormality is an autoimmune
`immunoregulatory
`erythematosis, chronic
`systemic lupus
`of:
`consisting
`the group
`selected from
`disease,
`bowel
`inflammatory
`diabetes mellitus,
`I
`type
`rheumatoid arthritis,
`ulcerative
`colitis, bullous
`disease,
`Crohn's
`sclerosis,
`cirrhosis, uveitis, multiple
`sarcoidosis, psoriasis, autoimmune myositis, Wegener's granulomatosis,
`pemphigoid,
`asthma.
`ichthyosis. Graves ophthalmopathy and
`
`biliary
`
`Claim 1 wherein the
`to
`The method according
`13.
`transplant
`marrow or organ
`bone
`is
`abnormality
`immunoregulatory
`versus-host disease.
`
`rejection
`
`or
`
`graft-
`
`brought
`
`lupus
`
`glomerulonephritis,
`
`diseases,
`
`Claim 1 wherein the
`to
`The method according
`14.
`from the group consisting of:
`selected
`is
`abnormality
`immunoregulatory
`diseases
`graft-versus-host
`tissue,
`or
`transplantation of organs
`systemic
`rheumatoid arthritis,
`including
`syndromes
`transplantation, autoimmune
`gravis, type I
`myasthenia
`sclerosis,
`erythematosus, Hashimoto's thyroiditis, multiple
`encephalomyelitis,
`allergic
`uveitis,
`diabetes, uveitis, posterior
`post-infectious
`and
`rheumatic fever
`including
`diseases
`post-infectious autoimmune
`skin
`hyperproliferative
`and
`inflammatory
`glomerulonephritis,
`seborrhoeic dermatitis,
`dermatitis,
`eczematous
`dermatitis,
`contact
`atopic dermatitis,
`urticaria,
`epidermolysis bullosa,
`pemphigoid,
`bullous
`lichen planus, pemphigus,
`acne,
`lupus erythematosus,
`eosinophilia,
`cutaneous
`erythema,
`angioedemas, vasculitis,
`associated
`uveitis
`conjunctivitis,
`vernal
`alopecia areata, keratoconjunctivitis,
`epithelialis
`cornea, dystrophia
`conical
`Behcet's disease, keratitis, herpetic keratitis,
`Graves'
`scleritis.
`Mooren's ulcer,
`pemphigus,
`ocular
`leukoma,
`comeae, corneal
`sarcoidosis,
`syndrome,
`opthalmopathy, Vogt-Koyanagi-Harada
`
`pollen
`
`-183-
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`SUN - IPR2017-01929, Ex. 1014, p. 657 of 970
`
`

`

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`
`PCT/US03/01120
`
`asthma,
`damage
`inflammatory
`bums,
`disease,
`
`multiple
`
`pure
`
`nodosa,
`
`5
`
`15
`
`20
`
`25
`
`30
`
`intrinsic
`asthma,
`allergic
`bronchial asthma,
`disease,
`airway
`reversible obstructive
`or inveterate
`asthma and
`chronic
`asthma,
`dust
`asthma,
`asthma, extrinsic
`vascular
`caused by
`ulcers,
`gastric
`bronchitis,
`airway hyper-responsiveness,
`diseases,
`bowel
`ischemic
`thrombosis,
`and
`ischemic diseases
`thermal
`associated with
`intestinal lesions
`enterocolitis,
`diseases, necrotizing
`Crohn's
`proctitis, eosinophilic gastroenteritis, mastocytosis,
`coeliac diseases,
`Goodpasture's
`nephritis,
`eczema, interstitial
`rhinitis,
`ulcerative colitis, migraine,
`nephropathy,
`myositis,
`diabetic
`syndrome,
`syndrome, hemolytic-uremic
`multiple neuritis,
`polyneuritis,
`disease,
`syndrome, Meniere's
`Guillain-Barre
`disease,
`Basedow's
`hyperthyroidism,
`radiculopathy,
`10 mononeuritis,
`purpura,
`thrombocytopenic
`anemia,
`idiopathic
`hypoplastic
`anemia,
`aplasia, aplastic
`anemia,
`pernicious
`agranulocytosis,
`anemia,
`autoimmune hemolytic
`interstitial
`idiopathic
`fibroid lung,
`sarcoidosis,
`osteoporosis,
`anemia, anerythroplasia,
`photoallergic
`pneumonia, dermatomyositis, leukoderma vulgaris, ichthyosis vulgaris,
`leukemia,
`sensitivity, cutaneous T cell lymphoma, chronic lymphocytic
`myocardosis,
`polyarteritis
`syndrome,
`aortitis
`atherosclerosis,
`arteriosclerosis,
`adiposis,
`eosinophilic
`syndrome,
`Sjogren's
`granuloma,
`scleroderma, Wegener's
`substantia
`bone,
`alveolar
`periodontium,
`of gingiva,
`fascitis, lesions
`senilis
`alopecia
`by preventing epilation
`or
`alopecia
`pattern
`glomerulonephritis, male
`generation
`hair
`promoting
`and/or
`or providing hair germination
`ischemia-
`disease,
`Sezary's syndrome, Addison's
`and
`muscular dystrophy, pyoderma
`or
`transplantation
`upon
`preservation,
`occurs
`which
`of organs
`injury
`reperfusion
`by drug
`colitis
`colitis,
`pseudomembranous
`ischemic disease, endotoxin-shock,
`insufficiency,
`renal
`insufficiency, chronic
`renal
`or radiation, ischemic acute
`emphysema, cataracta,
`lung cancer, pulmonary
`caused by lung-oxygen or drugs,
`corneal
`scarring,
`degeneration, vitreal
`macular
`senile
`pigmentosa,
`siderosis, retinitis
`IgA ballous dermatitis and cement
`linear
`alkali bum, dermatitis erythema multiforme,
`diseases
`by
`pancreatitis,
`sepsis,
`periodontitis,
`dermatitis, gingivitis,
`of
`,
`metastasis
`carcinogenesis,
`aging,
`environmental pollution,
`leukotriene-C4 release, Behcet's
`by histamine or
`caused
`hypobaropathy, disease
`sclerosing cholangitis,
`cirrhosis,
`biliary
`primary
`hepatitis,
`disease, autoimmune
`hepatitis,
`liver necrosis, necrosis caused by toxin, viral
`acute
`liver resection,
`cirrhosis, hepatic
`alcoholic
`hepatitis, cirrhosis,
`non-A/non-B
`anoxia, B-virus hepatitis,
`
`caused
`toxinosis
`
`caused
`carcinoma
`
`partial
`shock,
`
`or
`
`-184-
`
`SUN - IPR2017-01929, Ex. 1014, p. 658 of 970
`
`

`

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`
`PCT/US03/01120
`
`failure, fulminant
`liver
`failure, "acute-on-chronic"
`
`hepatic failure, late-onset hepatic
`
`HCMV
`
`failure, augmentation
`of
`chemotherapeutic
`effect, cytomegalovirus
`infection,
`infection, AIDS,
`
`cancer, senile dementia,
`trauma,
`
`and chronic bacterial
`infection.
`
`5
`
`15.
`immunoregulatory
`
`to Claim 1 wherein the
`The method according
`abnormality is multiple sclerosis.
`
`10
`
`16.
`immunoregulatory
`
`The method according to Claim 1 wherein the
`
`
`is abnormality rheumatoid arthritis.
`
`17.
`immunoregulatory
`
`The method according to Claim 1 wherein the
`
`abnormality is systemic lupus erythematosus.
`
`18.
`immunoregulatory
`
`The method according to Claim 1 wherein the
`
`
`is abnormality psoriasis.
`
`15
`
`19.
`immunoregulatory
`
`The method according to Claim 1 wherein the
`abnormality
`is
`rejection
`of
`
`transplanted
`
`organ
`
`20
`
`20.
`immunoregulatory
`
`The method according Claim 1 wherein the to
`
`
`
`
`is abnormality inflammatory bowel
`disease.
`
`25
`
`30
`
`21.
`immunoregulatory
`
`The method according
`to
`abnormality is a malignancy of
`
`Claim 1 wherein the
`
`lymphoid
`
`origin.
`
`22.
`immunoregulatory
`lymphomas.
`
`The method according to Claim 21 wherein the
`abnormality
`
`is and chronic acute
`
`
`lymphocytic leukemias and
`
`A pharmaceutical
`of
`comprised
`composition
`23.
`an
`in
`receptor
`the SIPi/Edgl
`is an agonist of
`compound
`said
`wherein
`abnormality,
`immunoregulatory
`SIPl/Edgl
`
`receptor
`
`the over S1PR3/Edg3
`receptor of at least 20
`fold
`
`as
`
`compound
`a
`amount
`
`possesses the
`measured
`
`by
`
`-185-
`
`SUN - IPR2017-01929, Ex. 1014, p. 659 of 970
`
`

`

`WO 03/061567
`
`PCT/US03/01120
`
`the to EC50 for the SlP3/Edg3
`
`
`receptor
`the for SIPi/Edgl
`
`
`the ratio of EC50
`as evaluated in the 35S-GTPYS binding
`
`assay and wherein said compound
`
`
`to an EC50 for binding the SIPi/Edgl
`receptor of 100
`nM
`or
`35S-GTPYS binding assay,
`
`receptor
`possesses
`less
`as evaluated
`
`5
`
`with the proviso
`
`that the compound
`
`does
`
`not
`
`fall
`
`within
`
`formula
`
`A:
`
`CH2R3
`O=P—X-CH2—C-CH2CH2^^-.
`N(R2)2
`
`R1A
`
`R1B
`
`A
`
`or a pharmaceutically
`
`acceptable
`
`salt
`
`10
`
`X is O, S, NRl
`
`or
`
`(CH2)l-2>
`
`Rl is H, Ci_4alkyl orhaloCi-4
`
`alkyl;
`
`
`
`hydrate or thereof, wherein:
`
`
`
`
`
`optionally 1-4 halo groups; substituted
`
`
`
`with
`
`Rla 1$ H, OH, Ci-4alkyl, or OCi_4 alkyl, the alkyl and
`
`substituted with 1-3 halo groups;
`
`15
`
`alkyl portions
`
`being
`
`optionally
`
`Rib
`
`represents H, OH, Ci-4 alkyl or haloCi_4
`
`alkyl;
`
`each R2 is independently selected from the group consisting of: H, Ci_4 alkyl and
`
`20
`
`haloCi-4 alkyl.
`
`R3 is H, OH, halo, Ci-4alkyl, OCi-4alkyl, 0-haloCi-4alkyl
`
`
`
`or hydroxyCi_4alkyl,
`
`-186-
`
`SUN - IPR2017-01929, Ex. 1014, p. 660 of 970
`
`

`

`WO 03/061567
`
`PCT/US03/01120
`
`Y is selected from the group consisting of: -CH2-, -C(O)-, -CH(OH)-, -C(=NOH)-, O
`
`and S, and
`
`5 R4 is selected from the group consisting of: C4_i4alkyl and C4_i4alkenyl,
`
`in combination with
`
`a
`
`pharmaceutically
`
`
`
`acceptable carrier.
`
`
`
`compound
`
`comprised
`A pharmaceutical composition
`24.
`is an agonist of
`the
`
`
`
`SIPi/Edgl in an amount effective receptor for treating an
`abnormality,
`wherein
`
`said possesses a selectivity for the compound
`
`immunoregulatory
`SIPl/Edgl
`receptor
`
`over the S1PR3/Edg3
`receptor of at least 100
`
`fold as measured by
`the ratio of EC50
`for the SIPl/Edgl
`
`
`receptor to the EC50 for the S IPs/EdgS receptor
`in the 35S-GTPYS binding
`as evaluated
`
`assay and wherein said
`
`compound possesses
`an EC50 for binding
`
`to the SIPl/Edgl
`receptor
`
`nM of 10 or less as evaluated by
`
`the
`35S-GTPYS binding
`
`assay, in combination with a
`
`pharmaceutically acceptable carrier.
`
`
`
`of which a
`
`10
`
`15
`
`20
`
`25
`
`25. A method of
`identifying a candidate compound which an
`is
`
`receptor
`
`that over the SlP3/Edg3 is
`
`selective receptor,
`
`agonist of the SIPi/Edgl
`wherein said
`candidate compound possesses
`a selectivity
`for SIPi/Edgl
`
`the receptor
`
`over the SlP3/Edg3
`receptor
`of at
`20 least fold as measured
`
`
`
`ratio the of EC50 for
`
`by
`the SIPl/Edgl receptor to
`the EC50 for
`the SlP3/Edg3
`
`as receptor evaluated in
`
`the
`SSS-GTPyS binding
`assay
`
`and wherein said candidate compound
`possesses
`an
`for binding
`to
`
`the SIPi/Edgl
`receptor
`of
`100 or less as evaluated nM
`
`
`by
`the
`35s-
`GTPyS binding assay.
`
`EC50
`
`with the proviso
`
`
`
`
`
`the that candidate compound
`
`
`
`does not fall within
`
`formula A:
`
`-187-
`
`SUN - IPR2017-01929, Ex. 1014, p. 661 of 970
`
`

`

`wo 03/061567
`
`pct/us03/01120
`
`R1A
`
`R1B
`
`CH2R3
`O=P—X-CH2—C-CH2CH2^^.
`N(R2)2
`
`V^Y-R4
`
`A
`
`or a pharmaceutically
`
`acceptable salt or hydrate thereof, wherein:
`
`5
`
`10
`
`15
`
`20
`
`X is O, S, NRl or
`
`(CH2)l-2> optionally
`
`Rl is H, Ci-4alkyl or haloCi_4 alkyl;
`
`the
`is H, OH, Ci-4alkyl, or OCi_4 alkyl,
`Rla
`substituted with 1-3
`halo
`groups;
`
`
`
`substituted halo groups; with
`
`
`
`
`
`alkyl alkyl portions being and
`
`
`
`1-4
`
`optionally
`
`Rib
`
`represents H, OH, Ci-4 alkyl or haloCi-4 alkyl;
`
`each R2
`
`
`
`is independently selected
`
`
`
`from the group consisting
`
`
`
`of: H, Ci_4 alkyl and
`
`haloCi_4 alkyl.
`
`R3 is H, OH, halo, Ci_4alkyl, OCi_4alkyl, 0-haloCi-4alkyl
`
`or
`
`hydroxyCi-4alkyl,
`
`Y is selected from the group consisting of: -CH2-, -C(O)-, -CH(OH)-, -C(=NOH)-, O
`and S, and
`
`R4 is selected from the group consisting of: C4_i4alkyl and C4_i4alkenyl,
`
`-188-
`
`SUN - IPR2017-01929, Ex. 1014, p. 662 of 970
`
`

`

`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`wo 03/061567
`
`comprising:
`
`pct/u s03/01120
`
`comprising:
`(1) providing a first receptor preparation
`receptor a functional or
`
`
`the SIPi/Edgl
`(a) a recombinant cell expressing
`
`equivalent of the SIPi/Edgl receptor capable of binding to
`sphingosine-1-
`phosphate ("SIP");
`or
`(b) a membrane preparation
`subsection (l)(a);
`
`of
`
`a
`
`
`
`recombinant in accordance with
`
`cell
`
`receptor preparation comprising:
`
`second
`(2) providing a
`a receptor or functional
`
`(a) a recombinant cell expressing
`the S IPs/EdgS
`
`
`equivalent of the SlP3/Edg3 receptor capable of binding to
`sphingosine-1-
`phosphate ("S IP"); or
`(b) a membrane preparation
`subsection (2)(a);
`
`of
`
`a
`
`
`
`recombinant accordance with cell
`
`
`
`in
`
`(3) separately contacting
`compound; and
`
`
`
`said cells or membrane
`
`preparations
`
`
`
`with candidate
`
`the
`
`activates
`and
`to
`candidate compound binds
`
`the
`(4) determining whether
`generated
`
`receptors
`by the level of a signal measuring
`
`SIPi/Edgl
`and SlP3/Ed3
`the interaction of
`the
`
`candidate compound with
`each
`receptor,
`thereby
`whether the candidate
`compound
`is
`an
`agonist
`of
`selective over
`the SlP3/Edg3 receptor.
`
`the
`
`the
`from
`indicating
`
`SIPi/Edgl
`
`26. A method in accordance
`with 25 wherein the method claim
`
`
`
`
`presence of labeled or unlabeled SIP, di-
`comprises conducting the method in
`the
`
`hydro SIP or a ligand for the SIPi/Edgl and/or SlP3/Edg3 receptor; provided
`that
`
`a ligand is utilized that is specific for either the SIPi/Edgl
`
`or SlP3/Edg3
`receptor, the
`receptor ligand utilized in the first receptor
`
`is preparation a ligand of the SIPi/Edgl
`
`receptor and
`the
`ligand
`
`
`
`
`in utilized second receptor the preparation
`is
`a
`ligand
`SlP3/Edg3
`receptor; and provided, further,
`
`that
`method the would additionally
`
`comprise measuring
`
`the level of a signal generated
`from
`the
`
`further
`
`if
`
`
`
`interaction di-
`
`of
`
`of
`
`-189-
`
`SUN - IPR2017-01929, Ex. 1014, p. 663 of 970
`
`

`

`wo 03/061567
`
`pct/us03/01120
`
`from
`signal
`the
`of
`effects a reduction
`that
`compound
`hydro SIP or ligand; wherein a
`the
`activates
`and
`receptor
`the
`SIP or ligand, with
`di-hydro
`SIP,
`the interaction of the
`obtained
`receptor is a
`that
`than
`level
`greater
`a
`at
`receptor
`SIPl/Edgl
`receptor.
`SIPl/Edgl
`the
`selective agonist of
`
`5
`
`A method in accordance
`27.
`by
`indicates extracellular pH changes caused
`
`with
`receptor
`
`claim
`activation.
`
`25
`
`10
`
`indicates
`
`accordance
`A method in
`28.
`within
`present
`levels of cAMP
`
`with
`
`claim
`the
`
`25
`cell.
`
`wherein
`
`wherein
`
`29.
`indicates adenylate
`
`accordance
`A method in
`accumulation.
`cyclase
`
`with
`
`claim
`
`25
`
`wherein
`
`15
`
`A method in accordance with
`30.
`flux.
`indicates Ca+
`
`claim
`
`25
`
`wherein
`
`25 wherein the candidate
`Claim
`to
`The method according
`31.
`SlP3/Edg3
`receptor
`the
`over
`receptor
`the
`SIPl/Edgl
`for
`selectivity
`compound has a
`the
`to
`receptor
`for
`SIPl/Edgl
`EC50
`of
`ratio
`the
`of at least 100 fold as measured by
`assay.
`SSs-GTPyS binding
`the
`in
`the EC50 for the SlP3/Edg3 receptor as evaluated
`
`candidate
`the
`Claim 31 wherein
`to
`The method according
`32.
`over the SlP3/Edg3 receptor
`receptor
`the SIPl/Edgl
`for
`selectivity
`a
`compound has
`receptor
`for the SIPj/Edgl
`by the ratio of EC50
`measured
`as
`fold
`of at least 200
`binding
`receptor as evaluated in the 35s-GTPyS
`the EC50 for the SlP3/Edg3
`
`to
`assay.
`
`wherein the candidate
`32
`Claim
`to
`The method according
`33.
`receptor
`the
`SlP3/Edg3
`over
`receptor
`the
`SIPl/Edgl
`for
`a selectivity
`compound has
`to
`receptor
`for the S IPi/Edgl
`the ratio of EC50
`by
`measured
`as
`fold
`of at least 500
`assay.
`the SSS-GTPyS binding
`in
`evaluated
`as
`receptor
`the EC50 for the SlP3/Edg3
`
`20
`
`25
`
`30
`
`-190-
`
`SUN - IPR2017-01929, Ex. 1014, p. 664 of 970
`
`

`

`wo 03/061567
`
`pct/us03/01120
`
`
`The method according Claim 33 wherein to
`the candidate
`
`
`34.
`the
`SlP3/Edg3 receptor
`
`selectivity for the SIPi/Edgl
`a
`receptor
`over
`
`compound has
`of at least 2000
`
`fold as measured by
`the
`ratio
`
`of for the SIPi/Edgl EC50
`
`receptor
`to
`
`the EC50 for the SlP3/Edg3 receptor as evaluated in the ^SS-GTPyS binding
`assay.
`
`The method according to Claim 25 wherein
`35.
`candidate
`the
`compound possesses
`an EC50 for binding to the SlPj/Edgl
`
`receptor of 1 nM or less
`as evaluated by
`the ^Ss-GTPyS binding
`
`assay.
`
`is
`36. A method of identifying a candidate compound which
`agonist of the SIPi/Edgl
`receptor
`that is selective over the SlP3/Edg3
`
`receptor,
`wherein said
`candidate
`compound
`possesses a selectivity
`
`for the SIPi/Edgl
`receptor
`over the SlP3/Edg3 receptor of at
`
`least 100 fold as measured by
`the
`
`ratio of EC50 for
`the SIPi/Edgl
`
`receptor to the EC50 for the SlP3/Edg3 receptor as evaluated in
`the
`35s-GTPyS binding assay and wherein
`
`said candidate compound
`
`possesses EC50 an
`
`for binding to the SlPj/Edgl
`receptor
`
`of nM or less as evaluated 10
`
`by
`the
`35s-
`GTPyS binding
`assay,
`comprising:
`
`an
`
`comprising:
`(1) providing a first receptor preparation
`receptor
`(a) a recombinant cell expressing the SIPi/Edgl
`equivalent of the SIPi/Edgl
`receptor
`phosphate ("SIP");
`or
`(b) a membrane
`preparation
`subsection (l)(a);
`
`of
`
`a
`
`recombinant
`
`or functional
`a
`
`
`capable sphingosine-1-
`
`of
`
`binding
`
`cell
`
`in
`
`(2) providing a
`receptor preparation comprising:
`
`second
`receptor or a functional
`(a) a recombinant cell
`
`expressing SlP3/Edg3 the
`
`equivalent of the SlP3/Edg3
`
`receptor capable of binding
`to
`sphingosine-1-
`phosphate ("SIP"); or
`
`(b) a membrane preparation a recombinant cell of
`subsection (2)(a);
`
`in
`
`accordance
`
`with
`
`
`
`-191-
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`SUN - IPR2017-01929, Ex. 1014, p. 665 of 970
`
`

`

`wo 03/061567
`
`pct/us03/01120
`
`(3) separately contacting said cells or membrane preparations with
`compound; and
`
`the
`
`candidate
`
`(4) determining whether the candidate compound
`5 SIPi/Edgl
`and SIPs/EdS
`receptors measuring
`
`the interaction of
`
`the candidate compound with
`whether the candidate compound
`is an agonist of
`selective over
`the SIPs/EdgS
`receptor.
`
`the
`to binds and activates
`
`
`signal generated from
`
`by the level of a
`
`each
`
`receptor, thereby indicating
`the
`SIPi/Edgl
`that is
`
`receptor
`
`10
`
`15
`
`36 the method further
`
`claim
`in accordance with
`37. A method
`
`
`labeled unlabeled SIP, di-or
`
`method in the presence of
`the
`comprises conducting
`
`hydro SIP or a ligand for the SIPi/Edgl
`that
`and/or
`
`SlP3/Edg3 receptor; provided
`a ligand is utilized
`that is specific for either the SIPi/Edgl
`or
`
`SlP3/Edg3 receptor, the
`
`receptor ligand utilized in the first receptor preparation
`is a ligand of the SIPi/Edgl
`receptor and
`
`the ligand utilized
`
`in the second receptor
`
`is preparation a ligand of the
`
`SlP3/Edg 3 receptor; and
`
`provided, further, that
`the method would additionally
`generated signal from the interaction of the SIP, di-
`comprise measuring
`the
`level
`of
`a
`
`
`
`
`
`
`a hydro SIP or ligand; wherein compound that effects reduction of the a
`
`signal
`from
`the interaction of
`the SIP,
`
`
`
`SIP di-hydro or ligand, with the
`receptor
`
`and the
`activates
`20 SIPl/Edgl
`receptor
`at
`a
`greater
`level
`than
`
`that receptor is a obtained
`
`selective agonist of
`the
`SIPl/Edgl
`receptor.
`
`wherein
`
`if
`
`A method in accordance with
`38.
`indicates extracellular
`pH
`
`changes caused by receptor
`
`claim
`activation.
`
`
`
`36 signal
`
`wherein
`
`25
`
`A method
`39.
`indicates levels of cAMP
`
`in accordance with
`present
`within
`
`claim
`the
`
`36 signal
`
`cell.
`
`wherein
`
`40.
`indicates adenylate
`
`A method in accordance with
`cyclase
`accumulation.
`
`30
`
`claim
`
`36
`
`wherein
`
`41. A method in accordance
`indicates Ca+
`flux.
`
`-192-
`
`
`
`with 36 wherein the signal claim
`
`
`
`SUN - IPR2017-01929, Ex. 1014, p. 666 of 970
`
`

`

`wo 03/061567
`
`pct/us03/01120
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`The method according
`42.
`selectivity
`
`for SIPi/Edgl
`compound has a
`of at least 200
`fold
`as measured
`the EC50 for the SIPs/EdgS
`receptor
`as
`
`wherein
`36
`Claim
`to
`receptor
`
`
`the receptor
`
`over SlP3/Edg3 the
`of receptor
`
`by
`
`the the SIPi/Edgl ratio
`
`assay.
`
`
`evaluated 35s-GTPyS binding in the
`
`
`
`
`EC50 to
`
`the
`
`to
`The method according
`43.
`compound has
`a selectivity
`for
`the
`
`of at least 500 fold as measured
`
`by ratio of EC50 the
`the EC50 for the SlP3/Edg3 receptor as evaluated
`
`the candidate
`Claim 42 wherein
`
`receptor
`
`SIPi/Edgl over the SlP3/Edg3
`receptor
`to
`
`for the SIPi/Edgl
`receptor
`
`in SSg-GTPyS binding the
`
`assay.
`
`The method according
`44.
`candidate
`to Claim 43 wherein
`
`SlP3/Edg3 receptor
`
`the receptor over
`the
`compound has
`a selectivity
`
`for SIPi/Edgl
`of at least 1000
`receptor
`to
`fold
`
`as measured ratio of EC50 by
`
`
`
`the the for SIPi/Edgl
`
`35S-GTPYS binding
`the EC50 for the S1PR3/Edg3 receptor as evaluated
`in
`the
`assay.
`
`the
`
`candidate
`the
`Claim 44 wherein
`
`to
`The method according
`45.
`
`receptor
`
`over SlP3/Edg3 the
`receptor
`
`
`for a selectivity the SIPi/Edgl
`compound has
`to
`
`
`fold
`as
`
`measured of EC50 for by
`the the SIPi/Edgl ratio
`
`receptor
`of at least 2000
`the EC50 for the SlP3/Edg3
`
`
`as receptor evaluated in
`the SSs-GTPyS
`binding
`
`assay.
`
`46.
`A method of treating
`disease condition in a
`
`respiratory
`a
`
`comprising said patient
`treatment
`such
`in
`need
`of
`mammalian patient
`an
`in
`receptor
`
`is an agonist of the SIPi/Edgl
`a compound which
`possesses
`compound
`a
`said
`treating said
`respiratory
`
`disease condition, wherein or
`
`selectivity for
`least
`20
`receptor
`of at
`the SIPi/Edgl
`receptor
`
`the over S1PR3/Edg3
`
`
`to receptor the EC50 for the
`
`fold as measured
`by
`the
`
`of EC50 ratio for the SlPj/Edgl
`
`SlP3/Edg3
`
`as receptor evaluated in
`
`the SSs-GTPyS binding
`assay
`and
`wherein
`compound possesses
`
`an EC50 for binding
`to the SIPi/Edgl
`
`receptor of 100 nM or
`
`
`the less as evaluated by SSs-GTPyS binding
`assay.
`
`or
`
`administering
`
`said
`
`with the proviso
`
`that
`
`the
`
`compound
`
`does
`
`
`
`not A:
`
`fall
`
`within
`
`-193-
`
`SUN - IPR2017-01929, Ex. 1014, p. 667 of 970
`
`

`

`WO 03/061567
`
`PCT/US03/01120
`
`R1A
`
`CHpR
`I
`O=P—X-CH2—C-CH2CH2
`
`R1b
`
`A
`
`4
`Y-R
`
`or a pharmaceutically acceptable salt or hydrate thereof, wherein:
`
`X is O, S, NRl or
`
`
`
`1-4 (032)1-2, optionally substituted with halo groups;
`
`
`
`
`
`5
`
`10
`
`15
`
`20
`
`Rl is H, Ci-4alkyl or haloCi-4
`
`alkyl;
`
`or OCi-4 alkyl, the alkyl and alkyl portions being optionally
`Rla is H, OH, Ci_4alkyl,
`substituted with
`1-3
`halo
`groups;
`
`Rib
`
`represents H, OH, Ci-4 alkyl or haloCi-4
`
`alkyl;
`
`is independently selected
`each R2
`haloCi-4 alkyl.
`
`from
`
`the
`
`
`
`group of: H, Ci-4 alkyl and consisting
`
`
`
`R3 is H, OH, halo, Ci-4alkyl,
`
`
`
`OCi-4alkyl, 0-haloCi_4alkyl
`
`or
`
`hydroxyCi-4alkyl,
`
`Y is selected
`and S, and
`
`
`
`from the group consisting of: -CH2-, -C(O)-,
`
`
`
`
`
`-CH(OH)-, -C(=NOH)-, O
`
`R4 is selected from
`
`the
`
`group
`
`consisting
`
`of:
`
`C4-I4alkyl
`
`and
`
`-194-
`
`SUN - IPR2017-01929, Ex. 1014, p. 668 of 970
`
`

`

`wo 03/061567
`
`pct/us03/01120
`
`47.
`The method according
`to
`46 wherein
`Claim
`the
`compound
`.
`a selectivity for
`the SlPj/Edgl
`receptor over
`SlP3/Edg3
`the
`receptor
`of at
`least
`fold as measured
`by
`the
`ratio
`of
`EC50
`for
`EC50 for
`the
`the
`35S-GTPYS binding
`SlP3/Edg3
`receptor
`as evaluated
`in
`the
`
`has
`
`100
`SIPi/Edgl
`assay.
`
`48.
`The method according
`to
`47 wherein
`Claim
`the
`compound
`has
`a selectivity for
`the SIPi/Edgl
`receptor over
`the
`receptor of at
`SlP3/Edg3
`least
`200
`fold as measured
`by
`the
`ratio
`the SIPi/Edgl
`of
`EC50
`receptor
`for
`EC50 for the
`to
`the
`SlP3/Edg3
`receptor as evaluated
`in
`the
`35s-GTPyS
`binding
`assay.
`
`5
`
`10
`
`49.
`The method according
`to
`Claim
`48
`wherein
`a selectivity for the SlPj/Edgl
`receptor
`SlP3/Edg3
`over
`the
`receptor of at least 500
`fold as measured by the
`of EC50 for
`ratio
`the SIPi/Edgl
`receptor
`the EC50 for the
`to
`15 SlP3/Edg3
`receptor as evaluated
`in the SSs-GTPyS binding
`assay.
`
`the
`
`50.
`The method according
`to
`wherein the compound
`Claim
`49
`has
`a selectivity for
`the SIPi/Edgl
`receptor
`over
`SlP3/Edg3
`the
`receptor of at least 2000
`fold as measured
`by
`the
`ratio
`for the SIPi/Edgl
`of
`EC50
`receptor
`EC50 for
`to
`the
`the
`20 SlP3/Edg3 receptor as evaluated
`in the SSg-OTPyS binding
`assay.
`
`51.
`A method of treating
`a
`respiratory
`or condition
`disease
`in
`a
`mammalian patient
`in
`need
`of
`such
`treatment
`comprising
`a compound which
`is
`an
`agonist
`of
`the
`in an amount effective
`SIPi/Edgl
`for
`treating said
`respiratory
`disease
`condition, wherein
`or
`said
`compound
`possesses
`selectivity for
`the SIPi/Edgl
`receptor
`over
`SlP3/Edg3
`the
`receptor of at least 100
`fold as measured
`by
`t