DocCode — SEQ.TXT
`
`SCORE Placeholder Sheet for IFW Content
`
`Application Number: 15467631
`
`Document Date: 03/23/2017
`
`The presence of this form in the IFW record indicates that the following document type was received
`in electronic format on the date identified above. This content is stored in the SCORE database.
`
`Since this was an electronic submission, there is no physical artifact folder, no artifact folder is
`recorded in PALM, and no paper documents or physical media exist. The TIFF images in the IFW
`record were created from the original documents that are stored in SCORE.
`
`• Sequence Listing
`
`At the time of document entry (noted above):
`• USPTO employees may access SCORE content via eDAN using the Supplemental Content
`tab, or via the SCORE web page.
`• External customers may access SCORE content via PAIR using the Supplemental Content
`tab.
`
`Form Revision Date: August 26, 2013
`
`0001
`
`MYLAN - EXHIBIT 1023
`
`

`

`PTO/AIA/15 (03-13)
`Approved for use through 01/31/2014. OMB 0651-0032
`U.S. Patent and Trademark Office. U.S. DEPARTMENT OF COMMERCE
`Under the Paperwork Reduction Act of 1995, no persons are required to respond to a collection of information unless it displays a valid OMB control number.
`SYPA-009/C03US
`
`Attorney Docket No.
`
`UTILITY
`PATENT APPLICATION
`TRANSMITTAL
`
`(Only for new nonprovisional applications under 37 CFR 1.53(b))
`
`Express Mail Label No.
`
`r
`
`K..
`
`First Inventor
`
`Stephen COMISKEY
`
`Title
`
`FORMULATIONS OF GUANYLATE
`CYCLASE C AGONISTS AND METHODS
`OF USE
`
`ADDRESS TO:
`
`Commissioner for Patents
`P.O. Box 1450
`Alexandria VA 22313-1450
`ACCOMPANYING APPLICATION PARTS
`
`10. q Assignment Papers
`(cover sheet & document(s))
`Name of Assignee
`
`11.
`
`q
`
`12.
`
`q
`
`13.
`
`q
`
`14.
`15.
`
`q
`
`16.
`
`q
`
`17.
`
`q
`
`18.
`
`q
`
`q Power of Attorney
`
`37 CFR 3.73(c) Statement
`(when there is an assignee)
`English Translation Document
`(if applicable)
`Information Disclosure Statement
`(PTO/SB/08 or PTO-1449)
`q Copies of citations attached
`
`Preliminary Amendment
`Return Receipt Postcard
`(MPEP 5 503) (Should be specifically itemized)
`Certified Copy of Priority Document(s)
`(if foreign priority is claimed)
`Nonpublication Request
`Under 35 U.S.C. 122(b)(2)(B)(i). Applicant must attach form
`PTO/SB/35 or equivalent.
`Other:
`
`APPLICATION ELEMENTS
`See MPEP chapter 600 concerning utility patent application contents.
`
`1.
`
`q
`
`2.
`
`3.
`
`q
`
`4.
`
`a
`
`Fee Transmittal Form
`(PTO/SB/17 or equivalent)
`Applicant asserts small entity status.
`See 37 CFR 1.27.
`Applicant certifies micro entity status. See 37 CFR 1.29.
`Applicant must attach form PTO/SB/15A or B or equivalent.
`[Total Pages 152
`]
`Specification
`Both the claims and abstract must start on a new page.
`(See MPEP § 608.01(a) for information on the preferred arrangement)
`[Total Sheets 6
`]
`Drawing(s). (35 U.S.C. 113)
`5.
`Oath or Declaration
`[Total Sheets
`]
`6. Inventor's
`substitute statements under 37 CFR 1.64 and assignments
`(including
`as an oath or declaration under 37 CFR 1.63(e))
`serving
`q Newly executed (original or copy)
`a.
`q A copy from a prior application (37 CFR 1.63(d))
`Application Data Sheet
`*See Note below.
`See 37 CFR 1.76 (PTO/AIA/14 or equivalent)
`CD-ROM or CD-R
`in duplicate, large table or Computer Program (Appendix)
`q Landscape Table on CD
`and/or Amino Acid Sequence Submission
`9. Nucleotide
`items a. — c. are required)
`
`(if applicable,
`Computer Readable Form (CRF)
`a.
`q
`Specification Sequence Listing on:
`b.
`q CD-ROM or CD-R (2 copies); or
`q Paper
`Statements verifying identity of above copies
`
`b.
`
`7.
`
`a
`
`8.
`
`q
`
`i.
`ii.
`q
`
`c.
`
`*Note:
`
`(1) Benefit claims under 37 CFR 1.78 and foreign priority claims under 1.55 must be included in an Application Data Sheet (ADS).
`(2) For applications filed under 35 U.S.C. 111, the application must contain an ADS specifying the applicant if the applicant is an
`assignee, person to whom the inventor is under an obligation to assign, or person who otherwise shows sufficient proprietary
`interest in the matter. See 37 CFR 1.46(b).
`19. CORRESPONDENCE ADDRESS
`
`0
`
`The address associated with Customer Number:
`
`58249
`
`OR
`
`q Correspondence address below
`
`Name
`
`Address
`
`City
`Country
`
`Signature
`Name
`Anne E. Fleckenstein
`
`(Print/Type) (Attorney/Agent)
`
`/Anne E. Fleckenstein/
`
`State
`Telephone
`
`Zip Code
`Email
`Date March 23, 2017
`Registration No. 62,951
`
`This collection of informa ion 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 12 minutes to complete,
`including gathering, preparing, and submitting the completed application form to the USPTO. Time 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.O. Box 1450, Alexandria, VA 22313-1450.
`If you need assistance in completing the form, call 1-800-PTO-9199 and select option 2.
`
`American LegalNet, Inc.
`www.Forms WofkFlow . coin
`
`143327320 v1
`
`0002
`
`

`

`Attorney Docket No.: SYPA-009/C03US 321994-
`
`PATENT
`
`IN THE UNITED STATES PATENT AND TRADEMARK OFFICE
`
`In Re Application of: Comiskey et al.
`
`Confirmation No.: To Be Assigned
`
`Serial No.: To Be Assigned
`(Continuation of U.S. Appl. No. 13/421,769)
`
`Group Art Unit:
`
`To Be Assigned
`
`Filed:
`
`herewith
`
`Examiner:
`
`To Be Assigned
`
`FOR:
`
`FORMULATIONS OF GUANYLATE CYCLASE C AGONISTS AND METHODS OF USE
`
`Mail Stop Amendment
`Commissioner for Patents
`P.O. Box 1450
`Alexandria, VA 22313-1450
`
`PRELIMINARY AMENDMENT
`
`Prior to examination of the above-identified application, please amend the above-
`
`captioned application as follows:
`
`Amendments to the Specification begin on page 2 of this paper.
`
`Amendments to the Claims begin on page 3 of this paper.
`
`Remarks begin on page 5 of this paper.
`
`143413675 vl
`
`1.
`
`0003
`
`

`

`Attorney Docket No.: SYPA-009/C03US 321994-
`(Continuation of U.S. Appl. No.: 13/421,769; Filed: March 15, 2012)
`
`IN THE SPECIFICATION:
`
`Please replace the paragraph under the heading "Related Applications" with the
`
`paragraph below. The amendments are indicated by strikethrough and underlining.
`
`This application is a continuation of U.S. Application No. 13/421,769, filed March 15,
`
`2012, which is a continuation-in-part of PCT/US2011/051805 filed on September 15, 2011,
`
`which claims the benefit of priority to U.S. Provisional Application No. 61/383,156 filed on
`
`September 15, 2010, U.S. Provisional Application No. 61/387,636 filed on September 29, 2010,
`
`and U.S. Provisional Application No. 61/392,186 filed on October 12, 2010, the contents of
`
`which are incorporated by reference in their entireties.
`
`Please insert the paragraph below at page 1, after the "Related Applications" section
`
`and before the "Field of Invention" section:
`
`INCORPORATION OF SEQUENCE LISTING
`
`The contents of the text file named "SYPA 009 CO3US SeqList ST25.txt", which was
`
`created on March 23, 2017 and is 113 KB in size, are hereby incorporated by reference in their
`
`entirety.
`
`143413675 vl
`
`2.
`
`0004
`
`

`

`Attorney Docket No.: SYPA-009/C03US 321994-
`(Continuation of U.S. Appl. No.: 13/421,769; Filed: March 15, 2012)
`
`IN THE CLAIMS:
`
`1-42. (Cancelled)
`
`43.
`
`(New) An oral dosage formulation of a Guanylate Cyclase-C (GCC) agonist peptide
`
`consisting of SEQ ID NO:1, wherein said peptide is a (4,12; 7,15) bicycle, an inert low
`
`moisture carrier and a lubricant, wherein the peptide has a chromatographic purity of no
`
`less than 91% after storage for at least three months.
`
`44.
`
`(New) The oral dosage formulation of claim 43, wherein the GCC agonist peptide has a
`
`chromatographic purity of no less than 92% to 95%.
`
`45.
`
`(New) The oral dosage formulation of claim 43, wherein the formulation contains less
`
`than 0.2% inorganic acids and carboxylic acids.
`
`46.
`
`(New) The oral dosage formulation of claim 43, wherein the formulation is a solid
`
`formulation and the unit dose is a powder, granule, sachet, troche, tablet, or capsule.
`
`47.
`
`(New) The oral dosage formulation of claim 43, wherein the GCC agonist peptide is
`
`stabilized against degradation for a period of at least 18 months at 30 °C and 65% relative
`
`humidity, or at least 18 months at 25 °C and 60% relative humidity, or at least 18 months
`
`at 2-8 °C.
`
`48.
`
`(New) The oral dosage formulation of claim 43, wherein the formulation is in the form of
`
`a capsule or tablet.
`
`49.
`
`(New) The oral dosage formulation of claim 48, wherein the capsule or tablet is in a
`
`blister pack or strip.
`
`50.
`
`(New) The oral dosage formulation of claim 43, wherein the lubricant is magnesium
`
`stearate.
`
`51.
`
`(New) The oral dosage formulation of claim 43, wherein the lubricant is at 0.25% (w/w).
`
`143413675 vl
`
`3.
`
`0005
`
`

`

`Attorney Docket No.: SYPA-009/C03US 321994-
`(Continuation of U.S. Appl. No.: 13/421,769; Filed: March 15, 2012)
`
`52.
`
`(New) The oral dosage formulation of claim 43, wherein the inert carrier is
`
`microcrystalline cellulose.
`
`53.
`
`(New) The oral dosage formulation of claim 43, wherein the inert carrier is at least 96%
`
`(w/w).
`
`54.
`
`(New) The oral dosage formulation of claim 43, wherein the inert carrier has a particle
`
`size of from 50 to 900 microns.
`
`143413675 vl
`
`4.
`
`0006
`
`

`

`Attorney Docket No.: SYPA-009/C03US 321994-
`(Continuation of U.S. Appl. No.: 13/421,769; Filed: March 15, 2012)
`
`Status of the Claims
`
`REMARKS
`
`Claims 43-54 are pending. Claims 1-42 are cancelled herein without prejudice or
`
`disclaimer. New claims 43-54 are added. Support for these new claims can be found throughout
`
`the application as filed, and specifically for example according to the table below. No new matter
`
`is added.
`
`Claim
`
`43
`
`44
`
`45
`
`46
`
`47
`
`48
`
`Support
`
`Claim
`
`Support
`
`Original claim 2, paragraphs [17]
`
`49
`
`Original claim 22
`
`and [41]
`
`Original claim 3
`
`Original claim 5
`
`Original claim 8
`
`Original claim 20
`
`Original claim 21
`
`50
`
`51
`
`52
`
`53
`
`54
`
`Paragraph [17]
`
`Paragraph [52]
`
`Original claim 10
`
`Paragraph [52]
`
`Original claim 11
`
`Entry and consideration of the foregoing amendments is respectfully requested. The
`
`specification has been amended to reflect the updated priority application and sequence listing
`
`information. No new matter is added.
`
`The Director is hereby authorized to charge any appropriate fees under 37 C.F.R. §§1.16,
`
`1.17, and 1.21 that may be required by this paper, and to credit any overpayment, to Deposit
`
`Account No. 50-1283.
`
`Dated: March 23, 2017
`
`COOLEY LLP
`ATTN: Patent Group
`1299 Pennsylvania Avenue NW, Suite 700
`Washington, DC 20004
`
`Tel: (202) 728-7030
`Fax: (202) 842-7899
`
`143413675 vl
`
`5.
`
`Respectfully submitted,
`COOLEY LLP
`
`By:
`
`/Anne E. Fleckenstein/
`Anne E Fleckenstein, Ph.D.
`Reg. No. 62,951
`
`0007
`
`

`

`Electronic Patent Application Fee Transmittal
`
`Application Number:
`
`Filing Date:
`
`Title of Invention :
`
`FORMULATIONS OF GUANYLATE CYCLASE C AGONISTS AND METHODS OF
`USE
`
`First Named Inventor/Applicant Name:
`
`Stephen COMISKEY
`
`Filer:
`
`Anne Elizabeth Fleckenstein
`
`Attorney Docket Number:
`
`SYPA-009C03US 321994-
`
`Filed as Small Entity
`
`Filing Fees for Utility under 35 USC 111(a)
`
`Description
`
`Fee Code
`
`Quantity
`
`Amount
`
`Sub-Total in
`USD($)
`
`Basic Filing:
`
`UTILITY FILING FEE (ELECTRONIC FILING)
`
`UTILITY SEARCH FEE
`
`UTILITY EXAMINATION FEE
`
`4011
`
`2111
`
`2311
`
`Pages:
`
`Claims:
`
`UTILITY APPL SIZE FEE PER 50 SHEETS >100
`
`2081
`
`Miscellaneous-Filing:
`
`Petition:
`
`1
`
`1
`
`1
`
`1
`
`70
`
`300
`
`360
`
`70
`
`300
`
`360
`
`200
`
`200
`
`0008
`
`

`

`Description
`
`Fee Code
`
`Quantity
`
`Amount
`
`Sub-Total in
`USD($)
`
`Patent-Appeals-and-Interference:
`
`Post-Allowance-and-Post-Issuance:
`
`Extension-of-Time:
`
`Miscellaneous:
`
`Total in USD ($)
`
`930
`
`0009
`
`

`

`Electronic Acknowledgement Receipt
`
`EFS ID:
`
`Application Number:
`
`28715843
`
`15467631
`
`International Application Number:
`
`Confirmation Number:
`
`9393
`
`Title of Invention :
`
`FORMULATIONS OF GUANYLATE CYCLASE C AGONISTS AND METHODS OF
`USE
`
`First Named Inventor/Applicant Name:
`
`Stephen COMISKEY
`
`Customer Number:
`
`58249
`
`Filer:
`
`Anne Elizabeth Fleckenstein
`
`Filer Authorized By:
`
`Attorney Docket Number:
`
`SYPA-009C03US 321994-
`
`Receipt Date:
`
`23-MAR-2017
`
`Filing Date:
`
`Time Stamp:
`
`16:22:59
`
`Application Type:
`
`Utility under 35 USC 111(a)
`
`Payment information:
`
`Submitted with Payment
`
`Payment Type
`
`Payment was successfully received in RAM
`
`yes
`
`DA
`
`$930
`
`RAM confirmation Number
`
`032417INTEFSW00016776501283
`
`Deposit Account
`
`Authorized User
`
`The Director of the USPTO is hereby authorized to charge indicated fees and credit any overpayment as follows:
`
`0010
`
`

`

`File Listing:
`
`Document
`Number
`
`1
`
`Document Description
`
`File Name
`
`File Size(Bytes)/
`Message Digest
`
`Multi
`Part /.zip
`
`Pages
`(if appl.)
`
`7214212
`
`SYPA_009_CO3US_Specificatio
`n.pdf
`
`12b4b4695a4a39d0220472e4e055f78a3k7
`993d
`
`yes
`
`158
`
`Multipart Description/PDF files in .zip description
`
`Document Description
`
`Start
`
`End
`
`Specification
`
`Claims
`
`Abstract
`
`Drawings-only black and white line drawings
`
`Warnings:
`
`Information:
`
`146
`
`151
`
`152
`
`158
`
`1
`
`147
`
`152
`
`153
`
`1793783
`
`2
`
`Application Data Sheet
`
`SYPA_009_CO3US_ADS.pdf
`
`no
`
`10
`
`7200d9f2c8afabf97b9dca8d9Ode841e4a
`448db
`
`230501
`
` _Transmittal.
`
`no
`
`1
`
`73aee30b157fc085793c9c99calfcee6f5a80
`40e
`
`fS
`
`
`
`SYPA_ 009_ p3dUCO
`
`
`
`Warnings:
`
`Information:
`
`3
`
`Transmittal of New Application
`
`Warnings:
`
`Information:
`
`4
`
`Sequence Listing (Text File)
`
`
`
`SeqList_
`
`SYPA_ 009_ CO3US
`
`5.txt
`
`
`
`ST2_
`
`115744
`
`no
`
`Warnings:
`
`Information:
`
`0011
`
`

`

`5
`
`Preliminary Amendment
`
`SYPA-009_CO3US_Prelim_Ame
`nd.pdf
`
`140794
`
`d24a1,53617287M39adc0d5b54e0b74d60
`a1267
`
`no
`
`5
`
`Warnings:
`
`Information:
`
`6
`
`Fee Worksheet (SB06)
`
`fee-info.pdf
`
`no
`
`2
`
`653c7d84W321a40c7c9Odaaa1123ae906
`30834
`
`36881
`
`Warnings:
`
`Information:
`
`Total Files Size (in bytes):
`
`9531915
`
`This Acknowledgement Receipt evidences receipt on the noted date by the USPTO of the indicated documents,
`characterized by the applicant, and including page counts, where applicable. It serves as evidence of receipt similar to a
`Post Card, as described in MPEP 503.
`
`New Applications Under 35 U.S.C. 111
`If a new application is being filed and the application includes the necessary components for a filing date (see 37 CFR
`1.53(b)-(d) and MPEP 506), a Filing Receipt (37 CFR 1.54) will be issued in due course and the date shown on this
`Acknowledgement Receipt will establish the filing date of the application.
`
`National Stage of an International Application under 35 U.S.C. 371
`If a timely submission to enter the national stage of an international application is compliant with the conditions of 35
`U.S.C. 371 and other applicable requirements a Form PCT/DO/EO/903 indicating acceptance of the application as a
`national stage submission under 35 U.S.C. 371 will be issued in addition to the Filing Receipt, in due course.
`
`New International Application Filed with the USPTO as a Receiving Office
`If a new international application is being filed and the international application includes the necessary components for
`an international filing date (see PCT Article 11 and MPEP 1810), a Notification of the International Application Number
`and of the International Filing Date (Form PCT/RO/105) will be issued in due course, subject to prescriptions concerning
`national security, and the date shown on this Acknowledgement Receipt will establish the international filing date of
`the application.
`
`0012
`
`

`

`Electronically Filed
`Date of Deposit: March 15, 2012
`
`Attny Ref.: 40737-509001US
`
`FORMULATIONS OF GUANYLATE CYCLASE C AGONISTS AND
`
`METHODS OF USE
`
`RELATED APPLICATIONS
`
`[01] This application is a continuation-in-part of PCT/US2011/051805 filed on September
`
`15, 2011, which claims the benefit of priority to U.S. Provisional Application No. 61/383,156
`
`filed on September 15, 2010, U.S. Provisional Application No. 61/387,636 filed on
`
`September 29, 2010, and U.S. Provisional Application No. 61/392,186 filed on October 12,
`
`2010, the contents of which are incorporated by reference in their entireties.
`
`FIELD OF THE INVENTION
`
`[02] The present invention relates to low-dose formulations of guanylate cyclase C peptide
`
`agonists useful for the treatment and prevention of various diseases and disorders.
`
`BACKGROUND OF THE INVENTION
`
`[03] Guanylate cyclase C is a transmembrane form of guanylate cyclase that is expressed
`
`on various cells, including gastrointestinal epithelial cells (reviewed in Vaandrager 2002 Mol.
`
`Cell. Biochem. 230:73-83). It was originally discovered as the intestinal receptor for the heat-
`
`stable toxin (ST) peptides secreted by enteric bacteria and which cause diarrhea. The ST
`
`peptides share a similar primary amino acid structure with two peptides isolated from
`
`intestinal mucosa and urine, guanylin and uroguanylin (Currie, et al., Proc. Nat'l Acad. Sci.
`
`USA 89:947-951 (1992); Hamra, et al., Proc. Nat'l Acad. Sci. USA 90:10464-10468 (1993);
`
`Forte, L., Reg. Pept. 81:25-39 (1999); Schulz, et al., Cell 63:941-948 (1990); Guba, et al.,
`
`Gastroenterology 111:1558-1568 (1996); Joo, et al., Am. J. Physiol. 274:G633-G644 (1998)).
`
`[04]
`
`In the intestines, guanylin and uroguanylin act as regulators of fluid and electrolyte
`
`balance. In response to high oral salt intake, these peptides are released into the intestinal
`
`lumen where they bind to guanylate cyclase C localized on the luminal membrane of
`
`enterocytes (simple columnar epithelial cells of the small intestines and colon). The binding
`
`of the guanylin peptides to guanylate cyclase C induces electrolyte and water excretion into
`
`1
`
`0013
`
`

`

`the intestinal lumen via a complex intracellular signaling cascade that is initiated by an
`
`increase in cyclic guanosine monophosphate (cGMP).
`
`[05] The cGMP-mediated signaling that is initiated by the guanylin peptides is critical for
`
`the normal functioning of the gut. Any abnormality in this process could lead to
`
`gastrointestinal disorders such as irritable bowel syndrome (IBS) and inflammatory bowel
`
`diseases. Inflammatory bowel disease is a general name given to a group of disorders that
`
`cause the intestines to become inflamed, characterized by red and swollen tissue. Examples
`
`include ulcerative colitis and Crohn's disease. Crohn's disease is a serious inflammatory
`
`disease that predominantly affects the ileum and colon, but can also occur in other sections of
`
`the gastrointestinal tract. Ulcerative colitis is exclusively an inflammatory disease of the
`
`colon, the large intestine. Unlike Crohn's disease, in which all layers of the intestine are
`
`involved, and in which there can be normal healthy bowel in between patches of diseased
`
`bowel, ulcerative colitis affects only the innermost lining (mucosa) of the colon in a
`
`continuous manner. Depending on which portion of the gastrointestinal tract is involved,
`
`Crohn's disease may be referred to as ileitis, regional enteritis, colitis, etc. Crohn's disease
`
`and ulcerative colitis differ from spastic colon or irritable bowel syndrome, which are
`
`motility disorders of the gastrointestinal tract. Gastrointestinal inflammation can be a chronic
`
`condition. It is estimated that as many as 1,000,000 Americans are afflicted with
`
`inflammatory bowel disease, with male and female patients appearing to be equally affected.
`
`Most cases are diagnosed before age 30, but the disease can occur in the sixth, seventh, and
`
`later decades of life.
`
`[06]
`
`IBS and chronic idiopathic constipation are pathological conditions that can cause a
`
`great deal of intestinal discomfort and distress but unlike the inflammatory bowel diseases,
`
`IBS does not cause the serious inflammation or changes in bowel tissue and it is not thought
`
`to increase the risk of colorectal cancer. In the past, inflammatory bowel disease, celiac
`
`disease and IBS were regarded as completely separate disorders. Now, with the description
`
`of inflammation, albeit low-grade, in IBS, and of symptom overlap between IBS and celiac
`
`disease, this contention has come under question. Acute bacterial gastroenteritis is the
`
`strongest risk factor identified to date for the subsequent development of postinfective
`
`irritable bowel syndrome. Clinical risk factors include prolonged acute illness and the
`
`absence of vomiting. A genetically determined susceptibility to inflammatory stimuli may
`
`also be a risk factor for irritable bowel syndrome. The underlying pathophysiology indicates
`2
`
`0014
`
`

`

`increased intestinal permeability and low-grade inflammation, as well as altered motility and
`
`visceral sensitivity. Serotonin (5-hydroxytryptamine [5-HT]) is a key modulator of gut
`
`function and is known to play a major role in pathophysiology of IBS. The activity of 5-HT is
`
`regulated by cGMP.
`
`[07] While the precise causes of IBS and inflammatory bowel diseases (IBD) are not
`
`known, a disruption in the process of continual renewal of the gastrointestinal mucosa may
`
`contribute to disease pathology in IBD and aggravate IBS. The renewal process of the
`
`gastrointestinal lining is an efficient and dynamic process involving the continual
`
`proliferation and replenishment of unwanted damaged cells. Proliferation rates of cells lining
`
`the gastrointestinal mucosa are very high, second only to the hematopoietic system.
`
`Gastrointestinal homeostasis depends on both the proliferation and programmed cellular
`
`death (apoptosis) of epithelial cells lining the gut mucosa. Cells are continually lost from the
`
`villus into the lumen of the gut and are replenished at a substantially equal rate by the
`
`proliferation of cells in the crypts, followed by their upward movement to the villus. The
`
`rates of cell proliferation and apoptosis in the gut epithelium can be increased or decreased in
`
`a variety of circumstances, e.g., in response to physiological stimuli such as aging,
`
`inflammatory signals, hormones, peptides, growth factors, chemicals and dietary habits. In
`
`addition, an enhanced proliferation rate is frequently associated with a reduction in turnover
`
`time and an expansion of the proliferative zone. The proliferation index is much higher in
`
`pathological states such as ulcerative colitis and other gastrointestinal disorders. Intestinal
`
`hyperplasia is a major promoter of gastrointestinal inflammation. Apoptosis and cell
`
`proliferation together regulate cell number and determine the proliferation index. Reduced
`
`rates of apoptosis are often associated with abnormal growth, inflammation, and neoplastic
`
`transformation. Thus, both increased proliferation and/or reduced cell death may increase the
`
`proliferation index of intestinal tissue, which may in turn lead to gastrointestinal
`
`inflammatory diseases.
`
`[08]
`
`In addition to a role for uroguanylin and guanylin as modulators of intestinal fluid and
`
`ion secretion, these peptides may also be involved in the continual renewal of gastrointestinal
`
`mucosa by maintaining the balance between proliferation and apoptosis. For example,
`
`uroguanylin and guanylin peptides appear to promote apoptosis by controlling cellular ion
`
`flux. Given the prevalence of inflammatory conditions in Western societies a need exists to
`
`3
`
`0015
`
`

`

`improve the treatment options for inflammatory conditions, particularly of the gastrointestinal
`
`tract.
`
`[09] Peptide agonists of guanylate cyclase C agonists ("GCC agonists") are described in
`
`U.S. Patent Nos. 7,041,786, 7,799,897, and U.S. Patent Application Publication Nos.
`
`US2009/0048175, US 2010/0069306, US 2010/0120694, US 2010/0093635, and US
`
`2010/0221329. However, the formulation of peptides for pharmaceutical delivery presents a
`
`number of special problems. For example, peptides are subject to structural modifications by
`
`a variety of degradation mechanisms resulting in problems of chemical and physical
`
`instability of the formulation.
`
`SUMMARY OF THE INVENTION
`
`[10] The present invention provides low-dose formulations of peptide agonists of
`
`guanylate cyclase C ("GCC") and methods for their use in the treatment and prevention of
`
`human diseases and disorders, such as a gastrointestinal motility disorder, irritable bowel
`
`syndrome, a functional gastrointestinal disorder, gastroesophageal reflux disease, functional
`
`heartburn, dyspepsia, functional dyspepsia, nonulcer dyspepsia, gastroparesis, chronic
`
`intestinal pseudo-obstruction, colonic pseudo-obstruction; Crohn's disease, ulcerative colitis,
`
`inflammatory bowel disease, colonic pseudo-obstruction, obesity, congestive heart failure,
`
`and benign prostatic hyperplasia. In certain embodiments, the formulations are stabilized
`
`against chemical degradation of the peptide. The low-dose formulations of the invention
`
`have unexpected efficacy in humans in a dosage range that was not predicted based on
`
`studies in primates. The formulations of the invention are particularly useful for the
`
`treatment or prevention of chronic idiopathic constipation. In certain embodiments, the GCC
`
`agonists are analogs of uroguanylin and bacterial ST peptides. In preferred embodiments, the
`
`analogs have superior properties compared to the naturally occurring or "wild-type" peptides.
`
`Examples of such superior properties include a high resistance to degradation at the N-
`
`terminus and C-terminus from carboxypeptidases, aminopeptidases, and/or by other
`
`proteolytic enzymes present in the stimulated human intestinal juices and human gastric
`
`juices. Examples of GCC agonists that can be used in the formulations and methods of the
`
`invention are described in more detail below and in U.S. Patent Nos. 7,041,786, 7,799,897,
`
`and U.S. Patent Application Publication Nos. US2009/0048175, US 2010/0069306, US
`
`4
`
`0016
`
`

`

`2010/0120694, US 2010/0093635, and US 2010/0221329, each of which is incorporated
`
`herein by reference in its entirety.
`
`[11] The invention provides an oral dosage formulation comprising one or more
`
`pharmaceutically acceptable excipients and at least one GCC agonist peptide, wherein the
`
`amount of GCC agonist peptide per unit dose is from 0.01 mg to 10 mg, and wherein the
`
`GCC agonist peptide is selected from the group consisting of SEQ ID NOs: 1-54 and 56-249.
`
`In one embodiment, the GCC agonist peptide has a chromatographic purity of no less than
`
`90%, no less than 90.5%, no less than 91%, no less than 92%, no less than 93%, no less than
`
`94%, no less than 95%, no less than 96%, no less than 97%, no less than 98%, or no less than
`
`99%. The chromatographic purity of the GCC agonist peptide is determined as area percent
`
`by HPLC. In one embodiment, the GCC agonist peptide is selected from the group
`
`consisting of SEQ ID NOs: 1, 8, 9, or 56. In one embodiment, the GCC agonist peptide is
`
`selected from the group consisting of SEQ ID NOs: 1 and 9. In one embodiment, the GCC
`
`agonist peptide is selected from the group consisting of SEQ ID NOs: 8 and 9. In one
`
`embodiment, the amount of GCC agonist peptide per unit dose is 0.1 mg, 0.3 mg, 0.6 mg, 1.0
`
`mg, 3.0 mg, 6.0 mg, 9.0 mg or 9.5 mg.
`
`[12]
`
`In one embodiment, the GCC agonist peptide has a total impurity content of no
`
`greater than 10%, no greater than 9.5%, no greater than 9%, no greater than 8%, no greater
`
`than 7%, no greater than 6%, no greater than 5%, no greater than 4%, no greater than 3%, no
`
`greater than 2%, or no greater than 1%. The total impurity content is determined as total area
`
`percentages of impurities by HPLC. The impurities do not include any pharmaceutically
`
`acceptable excipient used for the formulation. In one embodiment, the formulation is
`
`substantially free of inorganic acids and carboxylic acids, e.g., HC1, phosphoric acid, or
`
`acetic acid. In this context, carboxylic acids do not include amino acids or peptides. In this
`
`context "substantially" free of acids means that the acid content of the formulation at the time
`
`of packaging is preferably less than 0.2%, less than 0.1%, less than 0.05%, less than 0.01%,
`
`less than 0.005%, or less than 0.001% of the total weight of the formulation. In one
`
`embodiment, the formulation is free of HC1.
`
`[13]
`
`In one embodiment, the formulation is a solid formulation. In one embodiment, the
`
`formulation is in the form of a powder, granule, sachet, troche, tablet, or capsule. In another
`
`embodiment, the formulation is a liquid formulation and the GCC agonist peptide is in
`
`5
`
`0017
`
`

`

`solution or suspension in a lipophilic liquid. In one embodiment, the liquid is a refined
`
`specialty oil or a medium chain triglyceride or related ester. In one embodiment, the refined
`
`specialty oil is selected from Arachis oil, Castor oil, cottonseed oil, maize (corn) oil, olive oil,
`
`sesame oil, soybean oil, and sunflower oil. In one embodiment, the medium chain
`
`triglyceride or related ester is AKOMED E, AKOMED R, CAPTEX 355, LABRAFAC CC,
`
`LABRAFAC PG, LAUROGLYCOL FCC, MIGLYOL 810, MIGLYOL 812, MIGLYOL
`
`829, MIGLYOL 840, and SOFTISAN 645. In one embodiment, the liquid is selected from
`
`the group consisting of medium chain triglycerides, propylene glycol dicaprylocaprate,
`
`vitamin E, soybean oil, Cremaphor, PG, and PG 400. In one embodiment, the unit dose is a
`
`powder, tablet, or capsule. In one embodiment, the unit dose is a liquid-filled capsule. In one
`
`embodiment, the capsule or tablet is in a blister pack or strip. Preferably, the blister pack or
`
`strip is made of a material that is impermeable to water vapor and oxygen. In one
`
`embodiment the blister pack is comprised of a metal foil. In one embodiment the blister pack
`
`is a FOIL/FOIL blister pack. In one embodiment, the container of the blister pack is flushed
`
`with an inert gas such as nitrogen or argon. In one embodiment, the container further
`
`includes a desiccant. In a preferred embodiment the desiccant is a molecular sieve. In one
`
`embodiment, the unit dose is in a high density polyethylene bottle having a seal. In one
`
`embodiment, the bottle further comprises a desiccant. In one embodiment, the bottle further
`
`comprises an oxygen scavenger or molecular sieve. In one embodiment, the bottle is nearly
`
`impermeable to oxygen and water vapor (e.g., much more impermeable than a HDPE bottle),
`
`such as an OxyGuard bottle.
`
`[14]
`
`In one embodiment, the one or more pharmaceutically acceptable excipients include
`
`an inert carrier. In one embodiment, the inert carrier is a selected from mannitol, lactose, a
`
`microcrystalline cellulose, or starch. In one embodiment, the inert carrier has a particle size
`
`of from 50 to 900 microns, from 50 to 800 microns, from 50 to 300 microns, from 50 to 200
`
`microns, from 75 to 150 microns, from 75 to 200 microns, or from 75 to 300 microns.
`
`[15]
`
`In one embodiment, the GCC agonist peptide is stabilized against chemical or
`
`physical degradation for a period of at least 18 months at 30 °C and 65% relative humidity, or
`
`at least 18 months at 25 °C and 60% relative humidity, or at least 18 months at 2-8 °C.
`
`[16]
`
`In one embodiment, the one or more pharmaceutically acceptable excipients include a
`
`divalent cation salt such as calcium chloride. In one embodiment, the one or more
`
`6
`
`0018
`
`

`

`pharmaceutically acceptable excipients comprise an amino acid, such as leucine, histidine, or
`
`arginine, or an amine such TRIS or TRIS/HC1.
`
`[17]
`
`In one embodiment, the oral dosage formulation consists of the GCC agonist peptide
`
`described herein, an inert carrier (e.g., Celphere SCP-100, Avicel PH 102, or Avicel PH 112),
`
`and a lubricant (e.g., magnesium stearate). In one embodiment, the formulation consists of
`
`the GCC agonist peptide, an inert carrier (e.g., Avicel PH 200), a divalent cation salt (e.g.,
`
`calcium chloride or calcium ascorbate), an amino acid (e.g., leucine, histidine, or arginine) or
`
`a protective amine (e.g., TRIS), a coating agent (e.g., Methocel ES Premium LV) and
`
`optionally a lubricant (e.g., magnesium stearate) or another additive (e.g., trehalose). In one
`
`embodiment, the formulation consists of the GCC agonist peptide, a binder (e.g., Provsolv
`
`SMCC 90 LM), and a disintegrant (e.g., Explotab). In one embodiment, the formulation
`
`consists of the GCC agonist peptide, a diluent (e