UNITED STATES PATENT AND TRADEMARK OFFICE
`
`
`
`
`
`UNTTED STATES DEPARTMENT OF COMMERCE
`United States Patent and Trademark Office
`Address: COMMISSIONER FOR PATENTS
`PO. Box 1450
`Alexandria, Virginia 22313-1450
`WWW.uspto.gov
`
`
`
`APPLICATION
`FILING or
`GRP ART
`
`NUMBER
`371(c) DATE
`UNIT
`FIL FEE REC'D
`ATTY.DOCKET.NO
`61/392,186
`10/12/2010
`220
`40737-509P03US
`
`
`
`
`
`
`
`
`
`
`
`
`TOT CLAIMSQIND CLAIMS
`
`
`
`CONFIRMATION NO. 2241
`FILING RECEIPT
`LLL
`
`000000044204060
`
`30623
`MINTZ, LEVIN, COHN, FERRIS, GLOVSKY AND POPEO, P.C
`ONE FINANCIAL CENTER
`BOSTON, MA 02111
`
`Date Mailed: 10/28/2010
`
`Receipt is acknowledged of this provisional patent application. It will not be examined for patentability and will
`become abandoned not later than twelve months after its filing date. Any correspondence concerning the application
`must include the following identification information: the U.S. APPLICATION NUMBER, FILING DATE, NAME OF
`APPLICANT, and TITLE OF INVENTION. Fees transmitted by check or draft are subject to collection. Please verify
`the accuracy of the data presented on this receipt. If an error is noted on this Filing Receipt, please submit
`a written request for a Filing Receipt Correction. Please provide a copy of this Filing Receipt with the
`changes noted thereon. If you received a "Notice to File Missing Parts" for this application, please submit
`any corrections to this Filing Receipt with your reply to the Notice. When the USPTO processes the reply
`to the Notice, the USPTO will generate another Filing Receipt incorporating the requested corrections
`
`Applicant(s)
`
`Kunwar Shailubhai, Audubon, PA;
`
`Power of Attorney:
`Cynthia Kozakiewicz--42764
`
`If Required, Foreign Filing License Granted: 10/26/2010
`
`The country code and number of your priority application, to be used for filing abroad under the Paris Convention,
`is US 61/392,186
`
`Projected Publication Date: None, application is not eligible for pre-grant publication
`
`Non-Publication Request: No
`
`Early Publication Request: No
`Title
`
`Formulations Of Guanylate Cyclase C Agonists And Methods Of Use
`
`PROTECTING YOUR INVENTION OUTSIDE THE UNITED STATES
`
`Since the rights granted by a U.S. patent extend only throughout the territory of the United States and have no
`effect in a foreign country, an inventor who wishes patent protection in another country must apply for a patent
`in
`a specific country or in regional patent offices. Applicants may wish to consider the filing of an international
`application under the Patent Cooperation Treaty (PCT). An international (PCT) application generally has the same
`effect as a regular national patent application in each PCT-member country. The PCT process simplifies the filing
`of patent applications on the same invention in member countries, but does not result in a grant of "an international
`page 1 of 3
`
`
`
`0001
`
`MYLAN - EXHIBIT 1027
`
`

`

`patent” and does not eliminate the need of applicants to file additional documents and fees in countries where patent
`protection is desired.
`
`Almost every country has its own patent law, and a person desiring a patent in a particular country must make an
`application for patent in that country in accordance with its particular laws. Since the laws of many countries differ
`in various respects from the patent law of the United States, applicants are advised to seek guidance from specific
`foreign countries to ensure that patent rights are not lost prematurely.
`
`Applicants also are advised that in the case of inventions made in the United States, the Director of the USPTO must
`issue a license before applicants can apply for a patent in a foreign country. The filing of a U.S. patent application
`serves as a request for a foreign filing license. The application's filing receipt contains further information and
`guidance as to the status of applicant's license for foreign filing.
`
`Applicants may wish to consult the USPTO booklet, "General Information Concerning Patents” (specifically, the
`section entitled "Treaties and Foreign Patents") for more information on timeframes and deadlines for filing foreign
`patent applications. The guide is available either by contacting the USPTO Contact Center at 800-786-9199, or it
`can be viewed on the USPTO website at http://www .uspto.gov/web/offices/pac/doc/general/index.html.
`
`For information on preventing theft of your intellectual property (patents, trademarks and copyrights), you may wish
`to consult the U.S. Government website, http://www .stopfakes.gov. Part of a Department of Commerce initiative,
`this website includes self-help "toolkits" giving innovators guidance on how to protect intellectual property in specific
`countries such as China, Korea and Mexico. For questions regarding patent enforcement issues, applicants may
`call the U.S. Government hotline at 1-866-999-HALT (1-866-999-4158).
`
`LICENSE FOR FOREIGN FILING UNDER
`
`Title 35, United States Code, Section 184
`
`Title 37, Code of Federal Regulations, 5.11 & 5.15
`
`GRANTED
`
`The applicant has been granted a license under 35 U.S.C. 184, if the phrase "IF REQUIRED, FOREIGN FILING
`LICENSE GRANTED" followed by a date appears on this form. Such licenses are issued in all applications where
`the conditions for issuance of a license have been met, regardless of whether or not a license may be required as
`set forth in 37 CFR 5.15. The scope and limitations of this license are set forth in 37 CFR 5.15(a) unless an earlier
`license has been issued under 37 CFR 5.15(b). The license is subject to revocation upon written notification. The
`date indicated is the effective date of the license, unless an earlier license of similar scope has been granted under
`37 CFR 5.13 or 5.14.
`
`This license is to be retained by the licensee and may be used at any time on or after the effective date thereof unless
`it is revoked. This license is automatically transferred to any related applications(s) filed under 37 CFR 1.53(d). This
`license is not retroactive.
`
`The grant of a license does not in any way lessen the responsibility of a licensee for the security of the subject matter
`as imposed by any Government contract or the provisions of existing laws relating to espionage and the national
`security or the export of technical data. Licensees should apprise themselves of current regulations especially with
`respect to certain countries, of other agencies, particularly the Office of Defense Trade Controls, Department of
`State (with respect to Arms, Munitions and Implements of War (22 CFR 121-128)); the Bureau of Industry and
`page 2 of 3
`
`0002
`
`

`

`Security, Department of Commerce (15 CFR parts 730-774); the Office of Foreign AssetsControl, Department of
`Treasury (31 CFR Parts 500+) and the Department of Energy.
`
`NOT GRANTED
`
`No license under 35 U.S.C. 184 has been granted at this time, if the phrase "IF REQUIRED, FOREIGN FILING
`LICENSE GRANTED" DOES NOT appear on this form. Applicant may still petition for a license under 37 CFR 5.12,
`if a license is desired before the expiration of 6 months from the filing date of the application. If 6 months has lapsed
`from the filing date of this application and the licensee has not received any indication of a secrecy order under 35
`U.S.C. 181, the licensee may foreign file the application pursuant to 37 CFR 5.15(b).
`
`page 3 of 3
`
`0003
`
`

`

`=
`i
`
`ZZ Zz
`oil
`
`72%
`
`a
`
`%
`
`Z,
`=
`ragga
`Zr
`tis,
`> a
`#22
`(22 pr
`
`2 ==
`
`ZZ,
`2
`2
`Zor:
`A
`id
`Ed
`7%
`227
`Dad
`Fd
`
`Zot?
`
`ZZ;
`al
`fe]
`
`as
`E21
`Pe]
`pr
`% wr
`
`NN i
`
`hd 1
`FUE
`Coll
`
`Gprnen
`
`7 wm
`Sin
`
`Fig, 1
`
`35
`
`
`
`
`Time to first BM
`
`
`
`
`
`
`
`
`
`
`
`
`(say) sw
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`0004
`
`

`

`5.0 -
`
`4.0
`
`3.0 -
`
`1.0 -
`
`2.0 -
`
`0.0 -
`
`Fig. 2;
`
`Average
`rnuavber of weekly SBI
`
`Effect of daily treatment with plecanatide on spontaneoous bowel
`movements (SBM) in chronic constipation patients
`
` 7.0
`
`
`HM Pre-Tx
`
`6.0 -
`
`[Tx WK 1
`
`BTx Wk2
`
`
`Overall Tx
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`1.0mg
`
`3.0mg
`
`Treatment Groups
`
`
`
`0005
`
`

`

`Application Data Sheet
`
`
`
`Application Information:
`
`Application Type::
`
`Provisional
`
`Subject Matter::
`
`Suggested Group Art Unit:
`
`CD-ROM or CD-R?::
`
`Sequence submission?::
`
`Utility
`
`N/A
`
`None
`
`None
`
`Computer Readable Form (CRF)?::
`
`No
`
`Title::
`
`Formulations Of Guanylate Cyclase C Agonists
`
`Attorney Docket Number:
`
`40737-509P03US
`
`And Methods Of Use
`
`Request for Early Publication?::
`
`Request for Non-Publication?::
`
`Small Entity?::
`
`Petition included?::
`
`Secrecy Order in Parent Appl.?::
`
`No
`
`No
`
`No
`
`No
`
`No
`
`Applicant Information:
`
`Applicant Authority Type::
`
`Inventor
`
`Primary Citizenship Country:
`
`us
`
`Status:
`
`Given Name:
`
`Family Name::
`
`Full Capacity
`
`Kunwar
`
`Shailubhai
`
`Page # 1
`
`Initial 10/12/10
`
`0006
`
`

`

`City of Residence:
`
`Audubon
`
`State or Province of Residence:
`
`Country of Residence:
`
`PA
`
`us
`
`Street of mailing address:
`
`2707 Bald Eagle Circle
`
`City of mailing address::
`
`Audubon
`
`State or Province of mailing address:
`
`PA
`
`Postal or Zip Code of mailing address:
`
`19403
`
`Correspondence Information:
`
`Correspondence Customer Number:
`
`30623
`
`Representative Information:
`
`Representative Customer Number:
`
`30623
`
`Domestic Priority Information:
`
`Foreign Priority Information:
`
`Page # 2
`
`Initial 10/12/10
`
`0007
`
`

`

`Assignee Information:
`
`Synergy Pharmaceuticals, Inc.
`420 Lexington Avenue, Suite 1609
`New York, NY 10170
`
`Signature:
`
`/Cynthia Kozakiewicz/
`
`Date: October 12, 2010
`
`First: Cynthia
`
`Last: Kozakiewicz
`
`Reg. No.: 42,764
`
`5046701v.1
`
`Page # 3
`
`Initial 10/12/10
`
`0008
`
`

`

`Electronically Filed
`Date of Deposit: October 12, 2010
`
`Attny Ref.: 40737-509P03US
`
`FORMULATIONS OF GUANYLATE CYCLASE C AGONISTS AND
`
`METHODS OF USE
`
`RELATED APPLICATIONS
`
`[01]
`
`This application claims the benefit of priority to U.S. Provisional Application No.
`
`61/383,156 filed on September 15, 2010 and U.S. Provisional Application No. 61/387,63 filed on
`
`September 29, 2010 , the contents of which is 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, ef al., Proc. Nat'l Acad. Sci. USA
`
`89:947-951 (1992); Hamra, ef 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
`
`5031207v.1
`
`0009
`
`

`

`Attny Ref.: 40737-509P03US
`
`the guanylin peptides to guanylate cyclase C induces electrolyte and water excretion into 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
`
`2
`
`0010
`
`

`

`Attny Ref.: 40737-509P03US
`
`determined susceptibility to inflammatory stimuli may also be a risk factor for irritable bowel
`
`syndrome. The underlying pathophysiology indicates 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
`
`3
`
`0011
`
`

`

`Attny Ref.: 40737-509P03US
`
`ion flux. Given the prevalence of inflammatory conditions in Western societies a need exists to
`
`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
`
`4
`
`0012
`
`

`

`Attny Ref.: 40737-509P03US
`
`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, each of which is incorporated herein by reference in its entirety.
`
`[11]
`
`The invention provides an oral dosage formulation comprising at least one GCC
`
`agonist peptide, wherein the amount of GCC agonist peptide per unit dose is from 0.01 mg to 9.5
`
`mg, and wherein the GCC agonist peptide is selected from the group consisting of SEQ ID NOs:
`
`1-249. In one embodiment, the GCC agonist peptide is selected from the group consisting of
`
`SEQ ID NOs: 1, 8,9, 55 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 amount of GCC agonist
`
`peptide per unit dose is 0.1 mg, 0.3 mg, 1.0 mg, 3.0 mg, 9.0 mg or 9.5 mg.
`
`[12]
`
`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 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, and soybean oil.
`
`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 container of the blister pack is flushed with an inert gas such as nitrogen or argon. In one
`
`embodiment, the container further includes a dessicant. In a preferred embodiment the dessicant
`
`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 dessicant. In one embodiment,
`
`the bottle further comprises an oxygen scavenger or molecular sieve.
`
`5
`
`0013
`
`

`

`Attny Ref.: 40737-509P03US
`
`[13]
`
`In one embodiment, the formulation further comprises 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 200 microns, from
`
`75 to 150 microns, from 75 to 200 microns, or from 75 to 300 microns.
`
`[14]
`
`In one embodiment, the GCC agonist peptide is stabilized against chemical
`
`degradation for a period of at least 18 months at 25C and 60% relative humidity or at least 18
`
`months at 2-8C.
`
`[15]
`
`The invention also provides a process for making the oral dosage formulations
`
`described herein, wherein the process comprises a step of dry granulation or wet granulation. In
`
`another embodiment, the process comprises a step of dry mixing. In a preferred embodiment the
`
`step of dry mixing includes geometric blending. In one embodiment, the process comprises a
`
`step of direct compression.
`
`[16]
`
`The invention also provides a method for treating or preventing a disease or
`
`disorder in a subject in need thereof, comprising administering to the subject an oral dosage
`
`formulation comprising at least one GCC agonist peptide, wherein the amount of GCC agonist
`
`peptide per unit dose is from 0.01 mg to 9.5 mg, and wherein the GCC agonist peptide is selected
`
`from the group consisting of SEQ ID NOs: 1-249. Preferably, the subject is
`
`a human subject. In
`
`one embodiment, the GCC agonist peptide is selected from the group consisting of SEQ ID NOs:
`
`1, 8,9, 55 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 amount of GCC agonist peptide per
`
`unit dose is 0.1 mg, 0.3 mg, 1.0 mg, 3.0 mg, 9.0 mg or 9.5 mg.
`
`[17]
`
`In one embodiment, the disease or disorder is a gastrointestinal disease or disorder
`
`selected from the group consisting of irritable bowel syndrome, non-ulcer dyspepsia, chronic
`
`intestinal pseudo-obstruction, functional dyspepsia, colonic pseudo-obstruction, duodenogastric
`
`reflux, gastro esophageal reflux disease, constipation, gastroparesis, heartburn, gastric cancer,
`
`and H. pylori infection. In a preferred embodiment, the gastrointestinal disease or disorder is
`
`chronic idiopathic constipation.
`
`0014
`
`

`

`Attny Ref.: 40737-509P03US
`
`[18]
`
`In one embodiment, the method further comprises administering to the subject an
`
`effective amount of an inhibitor of a cGMP-specific phosphodiesterase. In one embodiment, the
`
`cGMP-dependent phosphodiesterase inhibitor is selected from the group consisting of suldinac
`
`sulfone, zaprinast, and motapizone, vardenifil, and suldenifil.
`
`[19]
`
`In one embodiment, the method further comprises administering to the subject an
`
`effective amount of at least one laxative. In one embodiment, the at least one laxative is selected
`
`from the group consisting of SENNA, MIRALAX, PEG, or calcium polycarbophil.
`
`[20]
`
`In one embodiment, the method further comprises administering to the subject an
`
`effective amount of at least one anti-inflammatory agent.
`
`[21]
`
`The invention also provides pharmaceutical compositions comprising the
`
`formulations described herein.
`
`[22]
`
`Other features and advantages of the invention will be apparent from and are
`
`encompassed by the following detailed description and claims.
`
`BRIEF DESCRIPTION OF THE DRAWINGS
`
`[23]
`
`dose.
`
`Figure 1: Plecanatide (SP-304) treatment reduced time to first BM following daily
`
`[24]
`
`Figure 2: Effect of daily treatment with plecanatide on spontaneous bowel
`
`movements (SBM) in chronic constipation patients.
`
`DETAILED DESCRIPTION
`
`[25]
`
`The invention provides pharmaceutical formulations of peptide GCC agonists. It
`
`is intended that the formulations of the invention are “pharmaceutical” formulations, meaning
`
`that they are suitable for pharmaceutical use. Accordingly, the term “formulations” as used
`
`herein is meant to encompass pharmaceutical formulations even if “pharmaceutical” is not
`
`expressly stated. Pharmaceutical compositions comprising the formulations described herein are
`
`also provided by the invention. The formulations of the invention preferably provide stability
`
`7
`
`0015
`
`

`

`Attny Ref.: 40737-509P03US
`
`against chemical and physical degradation of the peptide. The invention is based in part upon
`
`the discovery that mannitol mixes very effectively with the GCC agonist peptides described
`
`herein and provides stability against degradation, allowing the peptides to be formulated at very
`
`low doses. The invention is also based in part on the discovery that very low doses of the GCC
`
`agonist peptides described herein are effective for the treatment of diseases and disorders in
`
`humans. The dosage range found to be effective was not predicted based on animal studies.
`
`[26]
`
`The formulations of the invention are particularly useful for the treatment or
`
`prevention of a gastrointestinal disease or disorder selected from the group consisting of irritable
`
`bowel syndrome, non-ulcer dyspepsia, chronic intestinal pseudo-obstruction, functional
`
`dyspepsia, colonic pseudo-obstruction, duodenogastric reflux, gastro esophageal reflux disease,
`
`chronic idiopathic constipation, gastroparesis, heartburn, gastric cancer, and H. pylori infection.
`
`[27]
`
`In one embodiment, the formulations of the invention are used in a method for the
`
`treatment of constipation. Clinically accepted criteria that define constipation range from the
`
`frequency of bowel movements, the consistency of feces and the ease of bowel movement. One
`
`common definition of constipation is less than three bowel movements per week. Other
`
`definitions include abnormally hard stools or defecation that requires excessive straining.
`
`Constipation may be idiopathic (functional constipation or slow transit constipation) or
`
`secondary to other causes including neurologic, metabolic or endocrine disorders. These
`
`disorders include diabetes mellitus, hypothyroidism, hyperthyroidism, hypocalcaemia, Multiple
`
`sclerosis, Parkinson's disease, spinal cord lesions, Neurofibromatosis, autonomic neuropathy,
`
`Chagas disease, Hirschsprung disease and cystic fibrosis. Constipation may also be the result of
`
`surgery or due to the use of drugs such as analgesics (like opioids), antihypertensives,
`
`anticonvulsants, antidepressants, antispasmodics and antipsychotics. In a preferred embodiment,
`
`the constipation is chronic idiopathic constipation.
`
`[28]
`
`The stabilized formulations of the invention comprise at least one GCC agonist
`
`peptide formulated with one or more excipients such that the peptide is stabilized against
`
`chemical degradation. Chemical degradation of peptides results from a number of mechanisms
`
`including oxidation, water-mediated degradation, and reaction with aldheydes or reducing
`
`sugars. The ideal excipient or combination of excipients will be non-hygroscopic, have few or
`
`0016
`
`

`

`Attny Ref.: 40737-509P03US
`
`no reducing sugars, and be substantially free of contaminants such as iron, peroxide, and
`
`formaldehyde. The formulations of the invention are preferably substantially free of water. In
`
`this context “substantially” free of water means that the water content of the formulation at the
`
`time of packaging is preferably less than 7%, less than 5%, less than 1%, or less than 0.5% of the
`
`total weight of the formulation. In one embodiment the amount of water is between 0.05 to 5%
`
`of the total weight of the formulation.
`
`[29]
`
`In the context of the present formulations, the term “stable” or “stabilized” refers
`
`to the resistance of the peptide to chemical degradation over time. Preferably, a stable
`
`formulation of the invention retains an amount of the peptide in the formulation over a period of
`
`time that is at least 90%, preferably at least 95%, and most preferably at least 99% the amount of
`
`peptide initially present in the formulation. In one embodiment, the peptide is chemically stable
`
`in the formulation for a period of time that is at least 18 months, at least 20 months, or at least 24
`
`months when stored at 25 degrees Celsius (25C) and 60 % relative humidity. In one
`
`embodiment, the peptide is chemically stable in the formulation for a period of time that is at
`
`least 18 months, at least 20 months, or at least 24 months when stored at 2-8 degrees Celsius (2-
`
`8C). In one embodiment, the peptide is chemically stable in the formulation for a period of time
`
`that is at least 5 days, at least 7 days, at least 14 days, and preferably at least 30 days when stored
`
`at 25 degrees Celsius (25C) and 60 % relative humidity. In one embodiment, the peptide is
`
`chemically stable in the formulation for a period of time that is at least 5 days, at least 7 days, at
`
`least 14 days, and preferably at least 30 days when stored at 30 degrees Celsius (30C).
`
`[30]
`
`The low-dose formulations of the invention comprise an amount of at least one
`
`GCC agonist peptide per unit dose that is less than 10 mg. It is especially advantageous to
`
`formulate oral compositions in unit dosage form for ease of administration and uniformity of
`
`dosage. The term “unit dosage form” as used herein refers to physically discrete units suited as
`
`unitary dosages for the subject to be treated; each unit containing a predetermined quantity of
`
`active compound calculated to produce the desired therapeutic effect in association with the
`
`required pharmaceutical carrier. The specification for the dosage unit forms of the invention are
`
`dictated by and directly dependent on the unique characteristics of the active compound and the
`
`particular therapeutic effect to be achieved. In one embodiment, the unit dosage form is a tablet
`
`or a capsule.
`
`0017
`
`

`

`Attny Ref.: 40737-509P03US
`
`[31]
`
`In one embodiment of the low-dose formulations of the invention, the amount of
`
`GCC agonist peptide per unit dose is from 0.01 mg to 9.5 mg. In one embodiment, the amount
`
`of GCC agonist peptide per unit dose is 0.1 mg, 0.3 mg, 1.0 mg, 3.0 mg, 9.0 mg or 9.5 mg.
`
`[32]
`
`In one embodiment, the low-dose formulation contains a carrie