17667 U.S. PTO
`
`PTO/SB/13/PCT (01-05)
`1111111111111111111111111111111111111111
`Approved for use through 07/31/2006. 0MB 0651-0032
`051706
`U.S. Patent and Trademark Office; U.S. DEPARTMENT OF COMMERCE
`Under the Pa erwork Reduction Act or 1995, no ersons are re uired to res ond to a collection or information unless it dis la s a valid 0MB control number.
`REQUEST FOR FILING A CONTINUATION OR DIVISION OF AN INTERNATIONAL APPLICATION
`
`Pocket Numbe~
`
`Class
`
`ANTICIPATION CLASSIFICATION
`OF THIS APPLICATION
`I Subclass
`~683.2O4-USI
`Address to: Commissioner for Patents
`P.O. Box 1450
`Alexandria, VA 22313-1450
`
`PRIOR APPLICATION EXAMINER
`
`ART UNIT
`
`This is a request for filing a 00 continuation D divisional application under 37 CFR 1.53(b) of pending prior
`
`international application Number PCTDK2OO4/OOO792 • filed on November 18, 2004
`
`entitled PROPYLENE GLYCOL-CONTAINING PEPTIDE FORMULATIONS WIDCH ARE OPTIMAL
`FOR PRODUCTION AND FOR USE IN INJECTION DEVICES, which designated the United States.
`(5) TOTALS ($)
`(3) NUMBER EXTRA
`(4) RATE($)
`(1) FOR
`(2) NUMBER FILED
`
`TOTAL CLAIMS
`1441
`(37 CFR 1.16(i))
`INDEPENDENT
`[57
`CLAIMS 137 CFR 1.16/hll
`APPLICATION SIZE FEE (37 CFR 1.16(s))
`If the specification and drawings exceed 100 sheets of paper. the application size fee due
`is $250 ($125 for small entity) for each additional 50 sheets or fraction thereof. See 35
`U.S.C. 41(a)(1)(G) and 37 CFR 1.16/sl.
`
`-20 =
`
`-3=
`
`24
`
`2
`
`MULTIPLE DEPENDENT CLAIMS {37 CFR 1.16fin
`
`X
`
`X
`
`~
`lwol
`
`=
`
`=
`
`N/A
`
`BASIC FEE
`(37 CFR 1.16(a))
`SEARCH FEE
`(37 CFR 1.16(k))
`EXAMINATION FEE
`(37 CFR 1.16(0))
`
`1200.00
`
`400.00
`
`300.00
`
`500.00
`
`200.00
`
`Reduction by 50% for filing small entitv {Note 37 CFR 1.27)
`
`Total of above
`
`2600.00
`
`1. 00 Enclosed are the specification, claims and drawing(s).
`2. (cid:143) Applicant claims small entity status. See 37 CFR 1.27.
`
`Total
`
`2600.00
`
`3. [2fJ The Director is hereby authorized to charge any fees which may be required under 37 CFR 1.16 and 1.17. or credit any
`
`overpayment of Deposit Account No. 14-144 7 . A duplicate copy of this sheet is enclosed.
`
`4.
`
`A check in the amount of$ _______ is enclosed.
`
`Payment by credit card. Form PT0-2038 is attached.
`
`Application Data Sheet is enclosed. See 37 CFR 1.76.
`
`(cid:143)
`(cid:143) 5.
`6. ~
`
`7. ~
`If a CONTINUING APPLICATION, check appropriate box, and supply the requisite information below and in the first
`sentence of the specification following the title, or in an Application Data Sheet under 37 CFR 1. 76:
`00 Continuation D Divisional of prior PCT application No.: D1<2OO4/OOO792
`filed on November 18, 2004
`
`[Page 1 of2)
`This collection of information is required by 37 CFR 1.53(b). The information is required to obtain or retain a benefit by the public which is to file (and by the
`USPTO to process) an application. Confidentiality is governed by 35 U.S.C. 122 and 37 CFR 1.11 and 1.14. This collection is estimated to take 30 minutes to
`complete, including gathering, preparing, and submitting the completed application form to the USPTO. nme will vary depending upon the individual case. Any
`comments on the amount of time you require to complete this form and/or suggestions for reducing this burden, should be sent to the Chief Information Officer.
`U.S. Patent and Trademark Office, U.S. Department of Commerce, P.O. Box 1450, Alexandria, VA 22313-1450. DO NOT SEND FEES OR COMPLETED
`FORMS TO THIS ADDRESS. SEND TO: Commissioner for Patents, P .0. Box 1450, Alexandria, VA 22313-1450.
`/fyou need assistance in completing the form, call 1-800-PTO-9199 and select option 2.
`
` PFIZER, INC. v. NOVO NORDISK A/S - IPR2020-01252, Ex. 1003, p. 1 of 283
`
`

`

`PTO/SB/13/PCT (01-05)
`Approved for use through 07/31/2006. 0MB 0651-0032
`U.S. Patent and Trademark Office; U.S. DEPARTMENT OF COMMERCE
`Under the Paoerwork Reduction Act of 1995, no persons are reouired to respond to a collection of information unless it disolavs a valid 0MB control number.
`REQUEST FOR FILING A CONTINUING APPLICATION OF AN INTERNATIONAL
`APPLICATION
`8. [RI A declaration under CFR 1.63 is enclosed. (unsigned)
`
`9. [RI Priority offoreign application number PA 2003 01719, filed on November 20. 2003, in Denmark is claimed under 35 U.S.C.
`119(a)-(d).
`Priority of US application number60/524,653, filed on November 24, 2003, in the US is claimed under 35 U.S.C. 119(e)
`
`10.D A preliminary amendment is enclosed.
`
`11.D Also enclosed: _____________________________ _
`
`Address all future correspondence to: (May only be completed and signed by applicant, or attorney or agent of record).
`
`WARNING: Information on this form may become public. Credit card information should not
`be included on this form. Provide credit card information and authorization on PTO-2038.
`
`Signature
`
`Richard W. Bork
`Typed or printed name
`
`May 17, 2006
`
`Date
`
`Reg. No. 36,459
`Registration Number, if applicable
`
`(609) 987-5800
`Telephone Number
`
`(cid:143)
`(cid:143) Assignee of the entire interest. See 37 CFR 3.71. Statement under 37 CFR 3.73(b) is enclosed. (Form PTO/SB/96).
`(cid:143)
`
`lnventor(s)/Applicant(s)
`
`Attorney or agent of record
`
`[RI
`
`Filed under 37 CFR 1.34
`Registration number if acting under 37 CFR 1.34 36,459
`
`NOTE: Signatures of all the inventors or assignees of record of the entire interest or their representative(s) are required.
`Submit multiple forms if more than one signature is required, see below•.
`
`[RI "'Total of i forms are submitted.
`
`[Page 2 of 2]
`
` PFIZER, INC. v. NOVO NORDISK A/S - IPR2020-01252, Ex. 1003, p. 2 of 283
`
`

`

`Attorney Docket No.: 6683.204-US
`
`PATENT
`
`IN THE UNITED STATES PATENT AND TRADEMARK OFFICE
`EXPRESS MAIL CERTIFICATE
`
`Commissioner for Patents
`P.O. Box 1450
`Alexandria, VA 22313-1450
`
`Re: U.S. Patent Application for
`PEPTIDE
`GLYCOL-CONTAINING
`Title:
`PROPYLENE
`FORMULATIONS WHICH ARE OPTIMAL FOR PRODUCTION AND
`FOR USE IN INJECTION DEVICES
`Applicants: Pedersen et al.
`
`Sir:
`Express Mail Label No. EV 732210367 US
`Date of Deposit: May 17, 2006
`I hereby certify that the following attached paper(s) or fee
`
`1. Request for Filing a Continuation or Division of an International Application
`(in duplicate)
`2. Patent Application (34 pages of specification, 1 page of abstract,
`7 sheets of drawings)
`3. Unexecuted Combined Declaration and Power of Attorney
`4. Application Data Sheet (4 pages)
`5. Sequence Listing Transmittal Letter
`6. Sequence Listing (1 page)
`7. Computer Readable Format (CRF) of Sequence Listing
`are being deposited with the United States Postal Service "Express Mail Post Office to
`Addressee" under 37 C.F.R. 1. 10 on the date indicated above and is addressed to the
`Commissioner for Patents, PO Box 1450, Alexandria VA 22313-1450.
`
`Rashida Haji
`(Name of person mailing paper(s) or fee)
`.
`~~
`HCA) I'
`(Signature of person mailing paper(s) or fee)
`
`Mailing Address:
`Novo Nordisk Inc.
`Customer Number 23650
`
`Use the following customer number for all correspondence regarding this application.
`*23650*
`
`PATENT TRADEMARK OFFICE
`
` PFIZER, INC. v. NOVO NORDISK A/S - IPR2020-01252, Ex. 1003, p. 3 of 283
`
`

`

`Attorney Docket No.: 6683.204-US
`Express Mail Label No.: EV 732210367 US
`
`PROPYLENE GLYCOL-CONTAINING PEPTIDE FORMULATIONS WHICH ARE OPTIMAL
`
`FOR PRODUCTION AND FOR USE IN INJECTION DEVICES
`
`CROSS REFERENCE TO RELATED APPLICATIONS
`
`5
`
`This Application is a continuation of International Application serial no. PCT/DK2004/000792
`filed November 18, 2004 and claims priority from U.S. Application serial no. 60/524653 filed
`November 24, 2003 and from Danish Application serial no. PA 2003 01719 filed November
`
`20, 2003.
`
`10
`
`FIELD OF THE INVENTION
`
`15
`
`20
`
`25
`
`30
`
`The present invention relates to pharmaceutical formulations comprising a peptide
`and propylene glycol, to methods of preparing such formulations, and to uses of such formu(cid:173)
`lations in the treatment of diseases and conditions for which use of the peptide contained in
`such formulations is indicated. The present invention further relates to methods for reducing
`the clogging of injection devices by a peptide formulation and for reducing deposits on pro(cid:173)
`duction equipment during production of a peptide formulation.
`
`BACKGROUND OF THE INVENTION
`
`The inclusion of isotonicity agents in peptide-containing pharmaceutical formulations is widely
`known and one of the more common isotonic agents used in such formulations is mannitol.
`However, the present inventors have observed that mannitol causes problems during the pro(cid:173)
`duction of peptide formulations as it crystallizes resulting in deposits in the production equip(cid:173)
`ment and in the final product. Such deposits increase the need to clean the filling equipment
`during production of the formulation and this results in reduced production capability. In addi(cid:173)
`tion, such deposits may also result in reduced yield of the final product since vials/cartridges
`containing the peptide formulation may need to be discarded if particles are present. Finally, the
`present inventors have observed that in peptide formulations to be administered by injection, the
`presence of mannitol results in clogging of injection devices.
`
`Accordingly, it is desirable to identify an alternative isotonic agent to mannitol for inclusion in
`peptide-containing formulations and in particular, for inclusion in peptide formulations which are
`administered by injection.
`
`SUMMARY OF THE INVENTION
`The present inventors have discovered that peptide formulations containing propyl(cid:173)
`ene glycol at certain concentrations exhibit reduced deposits in production equipment and in
`the final product and also exhibit reduced clogging of injection devices. The present composi-
`
` PFIZER, INC. v. NOVO NORDISK A/S - IPR2020-01252, Ex. 1003, p. 4 of 283
`
`

`

`Attorney Docket No.: 6683.204-US
`Express Mail Label No.: EV 732210367 US
`
`2
`
`tions may be formulated with any peptide and are also physically and chemically stable thus
`
`rendering them shelf-stable and suitable for invasive (eg. injection, subcutaneous injection,
`
`intramuscular, intraveneous or infusion) as well as non-invasive (eg nasal, oral, pulmonary,
`
`transdermal or transmucosal e.g. buccal ) means of administration.
`
`5
`
`The present invention therefore relates to a pharmaceutical formulation comprising a
`
`peptide and propylene glycol, where the propylene glycol is present in a concentration of 1-
`
`100 mg/ml and the pH of the formulation is from 7-10. In a preferred embodiment, the phar(cid:173)
`
`maceutical formulations of the invention further contain a buffer and a preservative.
`
`The present invention also relates to methods for producing the pharmaceutical for-
`
`10 mutations of the invention.
`
`In one embodiment, the method for preparing a peptide formulation comprises:
`
`a)
`
`b)
`
`c)
`
`d)
`
`preparing a first solution by dissolving preservative, propylene glycol and
`
`buffer in water;
`
`preparing a second solution by dissolving the peptide in water;
`
`mixing the first and second solutions; and
`
`adjusting the pH of the mixture in c) to the desired pH.
`
`15
`
`In another embodiment, the method for preparing a peptide formulation comprises:
`
`a) preparing a first solution by dissolving preservative and buffer in water;
`
`20
`
`b) adding propylene glycol to the first solution;
`
`c) mixing the first solution with a second solution containing peptide dissolved in
`
`water; and
`
`d) adjusting the pH of the mixture inc) to the desired pH.
`
`25
`
`In yet another embodiment, the method for preparing a peptide formulation com(cid:173)
`
`prises:
`
`a) preparing a solution by dissolving preservative, buffer and propylene glycol in
`
`water;
`
`b) adding the peptide to the solution of step a); and
`
`30
`
`c) adjusting the pH of the solution of step b) to the desired pH.
`
`The present invention further relates to methods of treatment using the
`
`pharmaceutical formulations of the invention where the compositions are administered in an
`
`amount effective to combat the disease, condition, or disorder for which administration of the
`
`35
`
`peptide contained in the formulation is indicated.
`
` PFIZER, INC. v. NOVO NORDISK A/S - IPR2020-01252, Ex. 1003, p. 5 of 283
`
`

`

`Attorney Docket No.: 6683.204-US
`Express Mail Label No.: EV 732210367 US
`
`3
`
`In addition the present invention also relates to a method for reducing deposits on
`
`production equipment during production of a peptide formulation, where the method com(cid:173)
`
`prises replacing the isotonicity agent previously utilized in said formulation with propylene
`
`glycol at a concentration of between 1-100 mg/ml.
`
`5
`
`In one embodiment, the reduction in deposits on the production equipment during
`
`production by the propylene glycol-containing formulation relative to that observed for the
`
`formulation containing the previously utilized isotonicity agent is measured by a simulated
`
`filling experiment.
`
`The present invention also relates to a method for reducing deposits in the final
`
`10
`
`product during production of a peptide formulation, where the method comprises replacing
`
`the isotonicity agent previously utilized in said formulation with propylene glycol at a concen(cid:173)
`
`tration of between 1-100 mg/ml.
`
`In one embodiment, the reduction in deposits in the final product is measured by a
`
`reduction in the number of vials and/or cartridges of the propylene glycol-containing formula-
`
`15
`
`tion that must be discarded due to deposits relative to number of vials and/or cartridges of
`
`the formulation containing the previously utilized isotonicity agent that must be discarded due
`
`to deposits.
`
`The present invention further relates to a method for reducing the clogging of injec(cid:173)
`
`tion devices by a peptide formulation, where the method comprises replacing the isotonicity
`
`20
`
`agent previously utilized in said formulation with propylene glycol at a concentration of be(cid:173)
`
`tween 1-100 mg/ml.
`
`In one embodiment, the reduction in clogging of the injection device by the propyl(cid:173)
`
`ene glycol-containing formulation relative to that observed for the formulation containing the
`
`previously utilized isotonicity agent is measured in a simulated in use study.
`
`25
`
`30
`
`BRIEF DESCRIPTION OF THE FIGURES
`
`Figure 1 shows a photograph of dried droplets on microscope slides of from left to right, pla(cid:173)
`
`cebo (no peptide) formulations containing no isotonic agent (e only water, preservative and
`
`buffer), mannitol, sorbitol, xylitol, sucrose or glycerol as the isotonic agent with the far right
`slide containing mannitol with peptide Arg34, Lys26(Ne-(y-Glu(N°-hexadecanoyl)))-GLP-1(7-
`37).
`
`Figure 2 shows light microscopy pictures of from left to right, some of the dried droplets of
`
`35
`
`placebo formulations containing mannitol, arginin, inositol or glycerol as the isotonic agent.
`
` PFIZER, INC. v. NOVO NORDISK A/S - IPR2020-01252, Ex. 1003, p. 6 of 283
`
`

`

`Attorney Docket No.: 6683.204-US
`Express Mail Label No.: EV 732210367 US
`
`4
`
`Figure 3 shows light microscopy pictures of clogged needles dosed with placebo formula(cid:173)
`
`tions containing myoinositol, maltose or glycerol as the isotonic agent.
`
`Figure 4 shows light microscopy pictures of deposits on needles dosed with placebo formula-
`
`5
`
`tions containing glycine, lactose or mannitol as the isotonic agent.
`
`Figure 5 shows filling equipment after 24 hours simulated filling with Arg34, Lys26(NE-(y(cid:173)
`Glu(Ncx-hexadecanoyl)) )-GLP-1 (7-37) medium containing myo-inositol.
`
`1 o Figure 6 shows deposits on filling equipment after 24 hours simulated filling with a mannitol(cid:173)
`containing placebo formulation.
`
`Figure 7 shows deposits on needles dosed with mannitol (top panel) and propylene glycol
`(bottom panel)-containing Arg34
`, Lys26(NE-(y-Glu(Na-hexadecanoyl)) )-GLP-1 (7-37) formula-
`
`15
`
`tions.
`
`DESCRIPTION OF THE INVENTION
`
`20
`
`The present invention relates to a pharmaceutical formulation comprising a peptide
`
`or a mixture of peptides and propylene glycol where the final concentration of propylene gly(cid:173)
`
`col in the formulation is 1-100 mg/ml and the pH of the formulation is in the range of from 7-
`
`10.
`
`The pharmaceutical formulations of the invention are found to be optimal for produc-
`
`25
`
`tion because they exhibit reduced deposits in production equipment relative to formulations
`
`containing other isotonicity agents as measured by the simulated filling studies described in
`
`the Examples. In addition, the pharmaceutical formulations of the invention are found to be
`
`optimal for use in injection devices because they exhibit reduced clogging of the injection de(cid:173)
`
`vices relative to formulations containing other isotonicity agents as measured by the simu-
`
`30
`
`lated in use studies described in the Examples.
`
`The formulations of the present invention may be formulated with any peptide where
`
`examples of such peptides include, but are not limited to, glucagon, human growth hormone
`
`(hGH), insulin, aprotinin, FactorVII, tissue plasminogen activator (TPA), FactorVlla, FFR(cid:173)
`
`FactorVlla, heparinase, ACTH, Heparin Binding Protein, corticotropin-releasing factor, angio-
`
`35
`
`tensin, calcitonin, glucagon-like peptide-1, glucagon-like peptide-2, insulin-like growth factor-1,
`
`insulin-like growth factor-2, fibroblast growth factors, gastric inhibitory peptide, growth hormone-
`
` PFIZER, INC. v. NOVO NORDISK A/S - IPR2020-01252, Ex. 1003, p. 7 of 283
`
`

`

`Attorney Docket No.: 6683.204-US
`Express Mail Label No.: EV 732210367 US
`
`5
`
`releasing factor, pituitary adenylate cyclase activating peptide, secretin, enterogastrin, soma(cid:173)
`
`tostatin, somatomedin, parathyroid hormone, thrombopoietin, erythropoietin, hypothalamic re(cid:173)
`
`leasing factors, prolactin, thyroid stimulating hormones, endorphins, enkephalins, vasopressin,
`
`oxytocin, opiods, OPP IV, interleukins, immunoglobulins, complement inhibitors, serine protease
`
`5
`
`inhibitors, cytokines, cytokine receptors, PDGF, tumor necrosis factors, tumor necrosis factors
`
`receptors, growth factors and analogues as well as derivatives thereof where each of these
`
`peptides constitutes an alternative embodiment of the present invention.
`
`In the present application, the designation "an analogue" is used to designate a peptide
`
`wherein one or more amino acid residues of the parent peptide have been substituted by an-
`
`10
`
`other amino acid residue and/or wherein one or more amino acid residues of the parent peptide
`
`have been deleted and/or wherein one or more amino acid residues have been added to the.
`
`parent peptide. Such addition can take place either at the N-terminal end or at the C-terminal
`
`end of the parent peptide or both. Typically " an analogue" is a peptide wherein 6 or less
`
`amino acids have been substituted and/or added and/or deleted from the parent peptide,
`
`15 more preferably a peptide wherein 3 or less amino acids have been substituted and/or added
`
`and/or deleted from the parent peptide, and most preferably, a peptide wherein one amino
`
`acid has been substituted and/or added and/or deleted from the parent peptide.
`
`In the present application, "a derivative" is used to designate a peptide or analogue
`
`thereof which is chemically modified by introducing an organic substituent e.g. ester, alkyl or
`
`20
`
`lipophilic functionalities, on one or more amino acid residues of the peptide or analogue
`
`thereof.
`
`In one embodiment, the peptide to be included in the formulation of the invention is a
`
`GLP-1 agonist where "a GLP-1 agonist" is understood to refer to any peptide which fully or
`
`partially activates the human GLP-1 receptor. In a preferred embodiment, the "GLP-1
`
`25
`
`agonist" is any peptide that binds to a GLP-1 receptor, preferably with an affinity constant
`(Ko) or a potency (EC50) of below 1 µM, e.g. below 100 nM as measured by methods known
`in the art (see e.g. WO 98/08871) and exhibits insulinotropic activity, where insulinotropic
`
`activity may be measured in vivo or in vitro assays known to those of ordinary skill in the art.
`
`For example, the GLP-1 agonist may be administered to an animal and the insulin
`
`30
`
`concentration measured over time.
`
`Methods for identifying GLP-1 agonists are described in WO 93/19175 (Novo Nord(cid:173)
`
`isk A/S) and examples of suitable GLP-1 analogues and derivatives which can be used ac(cid:173)
`
`cording to the present invention includes those referred to in WO 99/43705 (Novo Nordisk
`
`A/S}, WO 99/43706 (Novo Nordisk A/S), WO 99/43707 (Novo Nordisk A/S), WO 98/08871
`
`35
`
`(analogues with lipophilic substituent) and in WO 02/46227 (analogues fused to serum albu(cid:173)
`
`min or to Fe portion of an lg).(Novo Nordisk A/S), WO 99/43708 (Novo Nordisk A/S), WO
`
` PFIZER, INC. v. NOVO NORDISK A/S - IPR2020-01252, Ex. 1003, p. 8 of 283
`
`

`

`Attorney Docket No.: 6683.204-US
`Express Mail Label No.: EV 732210367 US
`
`6
`
`99/43341 (Novo Nordisk A/S), WO 87/06941 (The General Hospital Corporation), WO
`90/11296 (The General Hospital Corporation), WO 91/11457 (Buckley et al.), WO 98/43658
`(Eli Lilly & Co.), EP 0708179-A2 (Eli Lilly & Co.), EP 0699686-A2 (Eli Lilly & Co.), WO
`01/98331 (Eli Lilly & Co).
`In one embodiment, the GLP-1 agonist is selected from the group consisting of
`GLP-1(7-36)-amide, GLP-1(7-37), a GLP-1(7-36)-amide analogue, a GLP-1(7-37) analogue,
`or a derivative of any of these.
`In one embodiment, the GLP-1 agonist is a derivative of GLP-1(7-36)-amide, GLP-
`1(7-37), a GLP-1(7-36)-amide analogue or a GLP-1(7-37) analogue, which comprises a lipo-
`philic substituent.
`In this embodiment of the invention, the GLP-1 derivative preferably has three
`lipophilic substituents, more preferably two lipophilic substituents, and most preferably one
`lipophilic substituent attached to the parent peptide (ie GLP-1(7-36)-amide, GLP-1(7-37), a
`GLP-1(7-36)-amide analogue or a GLP-1(7-37) analogue), where each lipophilic
`substituent(s) preferably has 4-40 carbon atoms, more preferably 8-30 carbon atoms, even
`more preferably 8-25 carbon atoms, even more preferably 12-25 carbon atoms, and most
`preferably 14-18 carbon atoms.
`In one embodiment, the lipophilic substituent comprises a partially or completely
`hydrogenated cyclopentanophenathrene skeleton.
`In another embodiment, the lipophilic substituent is a straight-chain or branched alkyl
`
`5
`
`10
`
`15
`
`20
`
`group.
`
`In yet another embodiment, the lipophilic substituent is an acyl group of a straight-chain
`or branched fatty acid. Preferably, the lipophilic substituent is an acyl group having the formula
`CH3(CH2)nCO-, wherein n is an integer from 4 to 38, preferably an integer from 12 to 38, and
`25 most preferably is CH3(CH2h2CO-, CH3(CH2)14CO-, CH3(CH2)15CO-, CH3(CH2)1aCO-,
`CH3(CH2)20CO- and CH3(CH2h2CO-. In a more preferred embodiment, the lipophilic substituent
`is tetradecanoyl. In a most preferred embodiment, the lipophilic substituent is hexadecanoyl.
`In a further embodiment of the present invention, the lipophilic substituent has a group
`which is negatively charged such as a carboxylic acid group. For example, the lipophilic
`
`30
`
`35
`
`substituent may be an acyl group of a straight-chain or branched alkane a,ro-dicarboxylic acid of
`the formula HOOC( CH2)mCO-, wherein m is an integer from 4 to 38, preferably an integer from
`12 to 38, and most preferably is HOOC(CH2)14CO-, HOOC(CH2)15CO-, HOOC(CH2)1sCO-,
`HOOC(CH2)20CO- or HOOC(CH2)22CO-.
`In the GLP-1 derivatives of the invention, the lipophilic substituent( s) contain a
`functional group which can be attached to one of the following functional groups of an amino
`acid of the parent GLP-1 peptide:
`
` PFIZER, INC. v. NOVO NORDISK A/S - IPR2020-01252, Ex. 1003, p. 9 of 283
`
`

`

`Attorney Docket No.: 6683.204-US
`Express Mail Label No.: EV 732210367 US
`
`7
`
`(a) the amino group attached to the alpha-carbon of the N-terminal amino acid,
`
`(b) the carboxy group attached to the alpha-carbon of the C-terminal amino acid,
`(c) the epsilon-amino group of any Lys residue,
`
`(d) the carboxy group of the R group of any Asp and Glu residue,
`(e) the hydroxy group of the R group of any Tyr, Ser and Thr residue,
`
`(f) the amino group of the R group of any Trp, Asn, Gin, Arg, and His residue, or
`(g) the thiol group of the R group of any Cys residue.
`
`In one embodiment, a lipophilic substituent is attached to the carboxy group of the R
`group of any Asp and Glu residue.
`In another embodiment, a lipophilic substituent is attached to the carboxy group
`
`attached to the alpha-carbon of the C-terminal amino acid.
`In a most preferred embodiment, a lipophilic substituent is attached to the epsilon(cid:173)
`amino group of any Lys residue.
`In a preferred embodiment of the invention, the lipophilic substituent is attached to the
`
`parent GLP-1 peptide by means of a spacer. A spacer must contain at least two functional
`groups, one to attach to a functional group of the lipophilic substituent and the other to a
`functional group of the parent GLP-1 peptide.
`
`In one embodiment, the spacer is an amino acid residue except Cys or Met, or a
`dipeptide such as Gly-Lys. For purposes of the present invention, the phrase "a dipeptide such
`as Gly-Lys" means any combination of two amino acids except Cys or Met, preferably a
`
`dipeptide wherein the C-terminal amino acid residue is Lys, His or Trp, preferably Lys, and the
`N-terminal amino acid residue is Ala, Arg, Asp, Asn, Gly, Glu, Gin, lie, Leu, Val, Phe, Pro, Ser,
`
`Tyr, Thr, Lys, His and Trp. Preferably, an amino group of the parent peptide forms an amide
`bond with a carboxylic group of the amino acid residue or dipeptide spacer, and an amino group
`of the amino acid residue or dipeptide spacer forms an amide bond with a carboxyl group of the
`lipophilic substituent.
`Preferred spacers are lysyl, glutamyl, asparagyl, glycyl, beta-alanyl and gamma(cid:173)
`
`aminobutanoyl, each of which constitutes an individual embodiment. Most preferred spacers are
`glutamyl and beta-alanyl. When the spacer is Lys, Glu or Asp, the carboxyl group thereof may
`form an amide bond with an amino group of the amino acid residue, and the amino group
`
`thereof may form an amide bond with a carboxyl group of the lipophilic substituent. When Lys is
`
`used as the spacer, a further spacer may in some instances be inserted between the s-amino
`
`group of Lys and the lipophilic substituent. In one embodiment, such a further spacer is succinic
`
`acid which forms an amide bond with the s-amino group of Lys and with an amino group present
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`in the lipophilic substituent. In another embodiment such a further spacer is Glu or Asp which
`
`forms an amide bond with the s-amino group of Lys and another amide bond with a carboxyl
`
` PFIZER, INC. v. NOVO NORDISK A/S - IPR2020-01252, Ex. 1003, p. 10 of 283
`
`

`

`Attorney Docket No.: 6683.204-US
`Express Mail Label No.: EV 732210367 US
`
`8
`
`group present in the lipophilic substituent, that is, the lipophilic substituent is a N&-acylated lysine
`
`residue.
`
`In another embodiment, the spacer is an unbranched alkane a.,ro-dicarboxylic acid
`
`5
`
`group having from 1 to 7 methylene groups, which spacer forms a bridge between an amino
`group of the parent peptide and an amino group of the lipophilic substituent. Preferably, the
`spacer is succinic acid.
`In a further embodiment, the lipophilic substituent with the attached spacer is a group
`of the formula CH3(CH2)pNH-CO(CH2)qCO-, wherein p is an integer from 8 to 33, preferably from
`12 to 28 and q is an integer from 1 to 6, preferably 2.
`In a further embodiment, the lipophilic substituent with the attached spacer is a group
`of the formula CHJ(CH2)rCO-NHCH(COOH)(CH2hCO-, wherein r is an integer from 4 to 24,
`preferably from 10 to 24.
`In a further embodiment, the lipophilic substituent with the attached spacer is a group
`of the formula CH3(CH2)5CO-NHCH((CH2}2COOH)CO-, wherein s is an integer from 4 to 24,
`preferably from 10 to 24.
`In a further embodiment, the lipophilic substituent is a group of the formula
`COOH(CH2}1CO- wherein t is an integer from 6 to 24.
`In a further embodiment, the lipophilic substituent with the attached spacer is a group
`of the formula -NHCH(COOH)(CH2)4NH-CO(CH2)uCH3, wherein u is an integer from 8 to 18.
`In a further embodiment, the lipophilic substituent with the attached spacer is a group
`of the formula CH3(CH2)vCO-NH-(CH2)z-CO, wherein v is an integer from 4 to 24 and z is an
`integer from 1 to 6.
`In a further embodiment, the lipophilic substituent with the attached spacer is a group
`of the formula -NHCH(COOH)(CH2)4NH-COCH((CH2hCOOH)NH-CO(CH2)wCH3, wherein w is
`an integer from 10 to 16.
`In a further embodiment, the lipophilic substituent with the attached spacer is a group
`of the formula -NHCH(COOH)(CH2)4NH-CO(CH2)2CH(COOH)NHCO(CH2)xCH3, wherein x is
`zero or an integer from 1 to 22, preferably 1 O to 16.
`In yet another embodiment the GLP-1 agonist is Arg34
`canoyl)))-GLP-1 (7-37).
`In yet another embodiment the GLP-1 agonist is selected from the group consisting
`of GI/-GLP-1 (7-36)-amide, Gly8-GLP-1 (7-37), Vals-GLP-1 (7-36)-amide, Vais ·GLP-1 (7-37),
`Vais Asp22-GLP-1 (7-36)-amide, Vais Asp22-GLP-1 (7-37) , ValsGlu22-GLP-1 (7-36)-amide ,
`ValsGlu22-GLP-1 (7-37), Valslys22-GLP-1 (7-36)-amide, Valslys22-GLP-1 (7-37), Vais Arg22-
`35 GLP-1 (7-36)-amide, Vais Arg22-GLP-1 (7-37), Val8His22-GLP-1 (7-36)-amide, ValsHis22-GLP-
`1 (7-37), analogues thereof and derivatives of any of these.
`
`10
`
`15
`
`20
`
`25
`
`30
`
`, Lys26(NE•(y-Glu(Na-hexade-
`
` PFIZER, INC. v. NOVO NORDISK A/S - IPR2020-01252, Ex. 1003, p. 11 of 283
`
`

`

`Attorney Docket No.: 6683.204-US
`Express Mail Label No.: EV 732210367 US
`
`9
`
`·
`
`5
`
`In yet another embodiment the GLP-1 agonist is selected from the group consisting
`of Arg26-GLP-1 (7-37); Arg34-GLP-1 (7-37); Lys36-GLP-1 (7-37); Arg26
`34Lys36-GLP-1 (7-37);
`•
`34Lys40-GLP-1 (7-37); Arg26Lys36-GLP-1 (7-37); Arg34Lys36-GLP-1 (7-
`Arg26·34-GLP-1 (7-37); Arg26
`37); Val8Arg22-GLP-1(7-37); Met8Arg22-GLP-1(7-37);Gly8His22-GLP-1(7-37); Val8His22-GLP-
`1 (7-37); Met8His22-GLP-1 (7-37);His37 -GLP-1(7-37); Gly8-GLP-1 (7-37); Val8-GLP-1 (7-37);
`Met8-GLP-1(7-37);Gly8Asp22-GLP-1(7-37}; Val8Asp22-GLP-1(7-37); Met8Asp22-GLP-1(7-
`37);Gly8Glu22-GLP-1 (7-37); Val8Glu22-GLP-1 (7-37); Met8Glu22-GLP-1 (7-37); Gly8Lys22-GLP-
`1 (7-37); Val8Lys22-GLP-1 (7-37); Met8Lys22-GLP-1 (7-37); Gly8Arg22-GLP-1 (7-37);
`Val8Lys22His37-GLP-1 (7-37); Gly8Glu22His37-GLP-1 (7-37); Val8Glu22His37 -GLP-1 (7-37);
`10 Met8Glu22His37-GLP-1 (7-37);Gly8Lys22 His37-GLP-1 (7-37); Met8Lys22His37-GLP-1 (7-
`37);Gly8Arg22His37-GLP-1 (7-37); Val8Arg22His37-GLP-1(7-37); Met8Arg22His37-GLP-1(7-37);
`Gly8His22His37 -GLP-1 (7-37); Val8His22His37 -GLP-1 (7-37); Met8His22His37 -GLP-1 (7-37);
`Gly8His37-GLP-1 (7-37); ValaHis37-GLP-1 (7-37); Met8His37-GLP-1 (7-37);Gly8Asp22 His37-GLP-
`1 (7-37); Val8Asp22His37-GLP-1 (7-37); MetaAsp22His37-GLP-1(7-37); Arg26-GLP-1(7-36)-amide;
`Arg34-GLP-1 (7-36)-amide; Lys36-GLP-1 (7-36)-amide; Arg26·34Lys 36-GLP-1 (7-36)-amide; Arg26·34
`GLP-1 (7-36)-amide; Arg26·34Lys40-GLP-1(7-36)-amide; Arg26Lys36-GLP-1(7-36)-amide;
`Arg34Lys36-GLP-1 (7-36)-amide; Gly8-GLP-1 (7-36)-amide; Vala-GLP-1 (7-36)-amide; Meta-GLP-
`1 (7-36)-amide;Gly8 Asp22-GLP-1 (7-36)-amide; Gly8Glu22His37 -GLP-1 (7-36)-amide; Vala Asp22-
`GLP-1 (7-36)-amide; Meta Asp22-GLP-1 (7-36)-amide;Gly8Glu22-GLP-1 (7-36)-amide; ValaGlu22-
`20 GLP-1(7-36)-amide; Met8Glu22-GLP-1(7-36)-amide; Gly8Lys22-GLP-1(7-36)-amide; Val8Lys22-
`GLP-1 (7-36)-amide; Metalys22-GLP-1 (7-36)-amide; Gly8His22His37 -GLP-1 (7-36)-amide;
`Gly8Arg22-GLP-1(7-36)-amide; Val8Arg22-GLP-1(7-36)-amide; Met8Arg 22-GLP-1(7-36)(cid:173)
`amide;Gly8His22-GLP-1(7-36)-amide; Val8His22-GLP-1(7-36)-amide; Met8His22-GLP-1(7-36)(cid:173)
`amide;His37 -GLP-1 (7-36)-amide; Vala Arg22His37 -GLP-1 (7-36)-amide; Meta Arg22His37 -GLP-
`1 (7-36)-amide; Gly8His37 -GLP-1 (7-36)-amide; Val8His37 -GLP-1 (7-36)-amide; Met8His37 -GLP-
`1 (7-36)-amide;Gly8 Asp22 His37 -GLP-1 (7-36)-amide; Vala Asp22His37 -GLP-1 (7-36)-amide;
`Met8 Asp22His37-GLP-1 (7-36)-amide; ValaGlu22His37-GLP-1 (7-36)-amide; Met8Glu22His37-GLP-
`1 (7-36)-amide;Gly8Lys22 His37 -GLP-1 (7-36)-amide; Va18Lys22His37-GLP-1 (7-36)-amide;
`Metalys22His37 -GLP-1 (7-36)-amide;Gly8 Arg22His37-GLP-1 (7-36)-amide; ValaHis22His37 -GLP-
`1 (7-36)-amide; MetaHis22His37 -GLP-1 {7-36)-amide; and derivatives thereof.
`In yet another embodiment the GLP-1 agonist is selected from the group consisting of
`Val8Trp 19Glu22-GLP-1 (7-37), ValaGlu22val25-GLP-1 (7-37), ValaTyr 16Glu22-GLP-1 (7-37),
`Val8Trp16Glu22-GLP-1(7-37), Val8Leu16Glu22-GLP-1{7-37), Val8Tyr1aGlu22-GLP-1(