AU2008201931A1 - Methods for treating LHRH-associated disorders with LHRH antagonists - Google Patents

Description

02-05-08;15:56 4/ -1-

AUSTRALIA

PATENTS ACT 1990 COMPLETE SPECIFICATION FOR A STANDARD PATENT

ORIGINAL

Name of Applicant/s: Actual Inventor/s: Praecis Pharmaceuticals Incorporated Marc B Garnick and Christopher J Molineaux and Malcolm L Gefter Address for Service is: SHELSTON IP Margaret Street SYDNEY NSW 2000 CCN: 3710000352 Attorney Code: SW Telephone No: Facsimile No.

(02) 97771111 (02) 9241 4666 Invention Title: METHODS FOR TREATING LRH ASSOCIATED DISORDERS WITH LHRH ANTAGONISTS Details of Original Application No. 2004203641 dated 04 Aug 2004 The following statement is a full description of this invention, including the best method of performing it known to me/us:- File: 32119AUP02 501539243_1.DOC/5844 COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 5/ -la- 00 0 SMETHODS FOR TREATING LRH ASSOCIATED DISORDERS WITH LHRH ANTAGONISTS ci Background of the Invention 5 Prostate cancer is a serious condition that affects increasing numbers of men O worldwide. About one-third of all men have at least some cancerous prostatic cells at age 50, with the incidence increasing to as many as 90 percent of men at age 90. In the United States alone, about 40,000 men die each year from prostate cancer.

Prostate cancer is a sex hormone dependent cancer; that is, the growth of the cancer is promoted by male hormones androgens such as testosterone and 0 dihydrotestosterone). Removal of the testes (castration) was for many years the standard 00 method of preventing the secretion of male hormones by the gonads, as a means for Sreducing growth of the cancer. More recently, secretion of male hormones has been Cl perturbed by chemical means by interfering with production of luteinizing hormone which regulates the synthesis of male hormones. Luteinizing hormone releasing hormone (LHRH) is a natural hormone produced by the hypothalamus that interacts with luteinizing hormone releasing hormone receptor (LHRH-R) in the pituitary to stimulate production of LH. To decrease LH production, superagonists of the luteinizing hormone releasing hormone receptor, such as leuprolide and goserelin, have been used. However, such LHRH superagonists initially act to stimulate LH release and only after prolonged treatment act to desensitize LHRH-R such that LH is no longer produced. The initial stimulation of LH production by the superagonist leads to an initial surge in the production of male hormones such that the initial response to superagonist therapy is aggravation, rather than amelioration, of the patient's condition tumor growth increases). This phenomenon, referred to herein as "agonist-induced hormone surge" or "agonist-induced testosterone surge" (also referred to in the art as the "flare reaction" or the "flare response"), can last for two to four weeks. Additionally, each successive administration of the superagonist can cause a small LH surge (known as the "acute-on chronic" phenomenon) that again can worsen the condition. The agonist-induced testosterone surge prohibits the use of LHRH superagonists in the treatment of late stage prostatic cancer patients where the cancer has metastasized to the spinal cord, since the initial stimulation of cancer growth would cause nerve trunk compression and damage.

To ensure that a candidate patient for superagonist therapy does not have spinal cord metastasis, additional diagnostic tests must be conducted, such as magnetic resonance imaging or a spinal CAT scan, which adds to the cost of superagonist therapy.

One approach that has been taken to avoid the agonist-induced testosterone surge has been to combine administration of an LHRH superagonist with an antiandrogen, such as flutamide, known as total androgen ablation therapy (AAT). Hormonal therapy COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08:15:56 6/ -2- 00 0 0 with an LHRH superagonist in combination with an antiandrogen has been used as a Cpre-treatment prior to radical prostatectomy, known as neoadjuvant therapy. The use of antiandrogens. however, is associated with serious hepatic and gastrointestinal side ieffects.

c Accordingly, methods for treating prostate cancer that are more effective than those utilizing LHRH superagonists, and that both avoid the occurrence of the agonistinduced testosterone surge and do not require the use of antiandrogens (thus avoiding the Sside-effects of using antiandrogens), are needed.

Summary of the Invention C1 The present invention features methods of treating prostate cancer designed to 00 Sreduce or eliminate the agonist-induced testosterone surge that occurs with current 0 prostate cancer therapies utilizing LHRH superagonists. Because the treatment methods of the invention avoid this testosterone surge, they are applicable to a wider number of prostatic cancer patients than is LHRH superagonist therapy the methods of the invention can be applied to patients with spinal cord metastasis). Moreover, certain expensive diagnostic tests that must be performed prior to initiating LHRH superagonist therapy may be eliminated when the methods of the invention are used an MRI or spinal CAT scan, which must be performed to rule out spinal cord metastasis before initiating LHRH superagonist therapy). Still further the methods of the invention can be performed without the use of an antiandrogen (although in certain optional embodiments an antiandrogen may be used) and therefore these methods can avoid the side-effects that occur with antiandrogen use.

The methods of the invention generally feature administration of an LHRH antagonist in combination with a second therapy. In one embodiment, this second therapy is a procedure to remove or destroy tumor tissue, such as a radical prostatectomy, cryosurgery or radiation therapy (external or interstitial). Preferably, the LHRH antagonist is administered to the subject prior to performing the procedure that removes or destroys prostatic tumor tissue. In a particularly preferred embodiment, the LHRH antagonist is administered to the subject for 3 to 6 months prior to performing the procedure that removes or destroys prostatic tumor tissue. In another embodiment of the methods of the invention, the second the! y is treatment with an LHRH agonist a superagonist. such as leuprolide, goserelin or decapeptyl). In this embodiment, the LHRH antagonist preferably is administered to the subject prior to initiating therapy with the LHRH agonist. Once LHRH agonist therapy has begun, LHRH antagonist therapy can be continued the antagonist and agonist can be coadministered) or discontinued first the LHRH antagonist is administered alone and then the LHRH agonist is administered alone). Moreover, the two-step combination methods of the COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15: 58 7/ -3- 00 O invention can further be combined with additional treatments, such as administration of an antiandrogen or administration of one or more inhibitors of sex steroid biosynthesis.

Another aspect of the invention pertains to methods for inhibiting agonistinduced hormone surge caused by LHRH agonist therapy in a subject in need of LHRH 5 agonist therapy, whatever the clinical setting prostate. ovarian or breast cancer o treatment, endometriosis, benign prostatic hypertrophy, and the like). In one embodiment, the method involves administering to the subject an LHRH antagonist in a sustained-release formulation; and administering to the subject an LHRH agonist, such Mn that the agonist-induced hormone surge is inhibited in the subject.

In another embodiment, the method involves administering to the subject an 0 LHRH antagonist for a period of at least about 14 days at a dose of about 15-300 00 ~g/kg/day (more preferably about 15-200 pg/kg/day and even more preferably about o 100 pg/kg/day); and administering to the subject an LHRH agonist, such that the cl agonist-induced hormone surge is inhibited in the subject In yet another embodiment, the method involves administering to the subject an LHRH antagonist having the following structure: Ac-D-Nal l 4-Cl-D-Phe 2 D-Pal 3

N-

Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-Alal 0 -LHRH; and administering to the subject an LHRH agonist, such that the agonist-induced hormone surge is inhibited in the subject.

In still another embodiment, the method involves: administering to the subject an LHRH antagonist having the following structure: Ac-D-Nall, 4-Cl-D-Phe 2 D-Pal 3 N-Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-Alao- LHRH at a dosage of about 15-300 pg/kg/day (more preferably about 15-200 pg/kg/day and even more preferably about 15-100 pg/kg/day) in a sustained-release formulation for a period of at least about 14 days; and administering to the subject an LHRH agonist; such that the agonist-induced hormone surge is inhibited in the subject.

Yet another aspect of the invention pertains to methods for inhibiting "acute-onchronic" agonist-induced hormone surge caused by periodic LHRH agonist therapy in a subject in need of LHRH agonist therapy, whatever the clinical setting prostate, ovarian or breast cancer treatment, endometriosis, benign prostatic hypertrophy, and the like). These methods involve: administering to the subject an LHRH agonist at regular intervals; and co-administering to the subject an LHRH antagonist with the LHRH agonist during each interval; such that the "acute-on-chronic" agonist-induced hormone surge is inhibited in the subject.

COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 -4- 00 0 Brief Description of the Drawings Figure I is a graphic representation of the plasma testosterone levels (in ng/ml) Sin adult male rats implanted with an osmotic pump releasing the LHRH antagonist PPI- 149 (at 15 or 50 pg/kg/day) for 28 days followed by removal of the PPI-149-releasing C, 5 pump and implantation of a second osmotic pump releasing the LHRH agonist o leuprolide (at 1.8 jpg/kg/day) such that there was no overlap in antagonist and agonist treatments.

Figure 2 is a graphic representation of the plasma testosterone levels (in ng/ml) in adult male rats implanted with an osmotic pump releasing the LHRH antagonist PPI- 149 (at 50 pg/kg/day) for 15 days, followed by implantation of a second osmotic pump 0,1 releasing the LHRH agonist leuprolide (at 1.8 pg/kg/day) on day 14 such that there was 00 one day of overlap in antagonist and agonist treatments.

O Figure 3 is a graphic representation of the plasma testosterone levels (in ng/mI) c in adult male rats implanted with an osmotic pump releasing the LHRH antagonist PPI- 149 (at 15 or 50 jg/kg/day) for 28 days, followed by implantation of a second osmotic pump releasing the LHRH agonist leuprolide (at 1.8 pg/kg/day) on day 27 such that there was one day of overlap in antagonist and agonist treatments.

Figure 4 is a graphic representation of the plasma testosterone levels (in ng/ml) in adult male rats implanted with an osmotic pump releasing the LHRH antagonist PPI- 149 (at 15 or 50 Ipg/kg/day) for 28 days, followed by implantation of a second osmotic pump releasing the LHRH agonist leuprolide (at 1.8 gg/kg/day) on day 21 such that there were seven days of overlap in antagonist and agonist treatments.

Figure 5 is a graphic representation of the plasma testosterone levels (in ng/ml) in cynomolgus monkeys implanted with an osmotic pump releasing the LHRH antagonist PPI1149 (at 0, 30, 100 or 300 pg/kg/day) for 28 days, followed by removal of the pump and administration of a single intramuscular dose of the depot, sustained release formulation of leuprolide equivalent to 1.6 mg/kg on day 28 such that there was no overlap in antagonist and agonist treatments.

Detailed Description of the Invention The present invention provides combination methods for treatment of prostate cancer in a subject in need thereof and compositions for use in such treatments.

As used herein, a "subject" is intended to include warm-blooded animals.

preferably mammals, including humans. In a preferred embodiment, the subject is a primate. In an even more preferred embodiment, the primate is a human.

The term "LHRH antagonist", as used herein, refers to a compound that inhibits the luteinizing hormone releasing hormone receptor such that release of luteinizing hormone is inhibited. The term "LHRH antagonist" may be used interchangeably with COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 G/ 00 O the term "LHRH-R antagonist" to refer to compounds that inhibit LHRH-R such that release of LH is inhibited. LHRH antagonists have been described in the art; see e.g., U.S. Patent 5,470,947 to Folkers er al.; Folkers et al., PCT Publication No. WO 89/01944; U.S. Patent 5,413,990 to Haviv; U.S. Patent 5,300,492 to Haviv; U.S Patent C 5 5,371,070 to Koerber et al.; U.S. Patent 5,296,468 to Hoeger et al.; U.S. Patent O 5,171,835 to Janaky et al.; U.S. Patent 5,003,011 to Coy et U.S. Patent 4,431.635 to Coy; U.S. Patent 4,992,421 to De et al.; U.S. Patent 4,851,385 to Roeske; U.S. Patent 4,801,577 to Nestor, Jr. et al.; and U.S. Patent 4,689,396 to Roeske et a. Preferred

S

E LHRH antagonists are those having low histamine-releasing activity an ED 50 for histamine release in a standard in vitro histamine release assay of at least 3 pg/ml, more 0 preferably at least 5 pg/ml, and still more preferably at least 10 pIg/ml) and that exhibit 00 water solubility. Preferred LHRH antagonists with low histamine-releasing activity and water solubility include compounds disclosed in U.S. Patent Application Serial No.

NC 08/480,494, filed on June 7, 1995, the entire contents of which is expressly incorporated herein by reference. An especially preferred LHRH antagonist comprises the structure: Ac-D-Nal1, 4-Cl-D-Phe 2 D-Pal 3 N-Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-Ala- LHRH (referred to herein as PPI-149)(described further in U.S. Patent Application Serial No.

08/480,494). Preferably, histamine-releasing activity is assayed by the method described in U.S. Patent 4,851,385 to Roeske. The efficacy of candidate LHRH antagonists in inhibiting LH release can be assayed, for example, in an animal model such as that described in Corbin and Beattie, Endocrine Res. Commun. 2:1 (1975). In this assay, the LHRH antagonistic activity of a candidate compound is assayed by measuring the antiovulatory activity (AOA) of the compound in rats.

The term "LHRH agonist", as used herein, refers to a compound that stimulates the luteinizing hormone releasing hormone receptor such that luteinizing hormone is released a compound that mimics the activity of LHRH). An LHRH agonist can have greater LH-releasing activity than natural LHRH (referred to as a "superagonist").

Many LHRH agonists and superagonists are known in the art. Commercially available LHRH agonists include leuprolide (trade name: Lupron®; Abbott/TAP), goserelin (trade name: Zoladex®; Zeneca), buserelin (Hoechst), triptorelin (also known as Decapeptyl, D-Trp-6-LHRH and Debiopharm; Ipsen/Beaufour), nafarelin (trade name" Synarel®; Syntex), lutrelin (Wyeth), cystorelin (Hoechst), gonadorelin (Ayerst) and histrelin (Ortho). Preferred LHRH agonists are leuprolide, goserelin and triptorelin.

For reviews of LHRH agonists and antagonists, see also B.H. Vickery et al., eds., (1984) "LHRH and Its Analogs: Contraceptive and Therapeutic Applications", MTP Press Limited. Lancaster, PA; and G. Schaison (1989) J. Steroid Biochem. 33(4B): 795.

Exemplary LHRH agonist and antagonists useful in the methods of the present invention COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 It 10/ -6- 00 O include nona- and decapeptides, as well as peptidomimetics, that mimic the structure of natural LHRH.

c An "antiandrogen", as used herein, refers to a compound that antagonizes the release or action of androgens. Antiandrogens are known in the art (see. U.S.

Patent 4,386.080), and are commercially available Androcur, a product of Schering O Candidate antiandrogens can be evaluated by methods known in the art (see, e.g., Goos et al, (1982) "An Improved Method of Evaluating Antiandrogens. Arch.

Dermatol. Res., 273:333-341). Antiandrogens can be steroidal or nonsteroidal.

Preferred antiandrogens for use in the methods of the invention include nonsteroidal antiandrogens such as flutamide (4'-nitro-3'-trifluorormethyl isobutyranilide; available 0S from Schering-Plough under the trade name Eulexin®), bicalutamide and nilutamide.

0 0 The term "inhibitor of sex steroid biosynthesis" is intended to include inhibitors O of adrenal sex steroid biosynthesis aminoglutethimide) and inhibitors of testicular sex steroid biosynthesis ketoconazole), or combinations thereof. When an inhibitor of adrenal sex steroid biosynthesis is employed, it may be desirable to simultaneously administer hydrocortisone to the patient in an amount sufficient to maintain normal glucocorticoid levels.

Various aspects of the invention are described further in the following subsections.

I. Methods for Treating Prostate Cancer The methods of the invention generally feature the administration of an LHRH antagonist in combination with a second therapy, such as performance of a procedure that removes or destroys tumor tissue or administration of an LHRH agonist.

One aspect of the invention pertains to a method for treating prostate cancer in a subject in need of such treatment, comprising administering to the subject an LHRH antagonist, and performing on the subject at least one procedure that removes or destroys prostatic tumor tissue, such as a radical prostatectomy, cryosurgery, external radiation therapy X-ray therapy) or interstitial radiation therapy implantation of a radioactive seed). The type, dosage and duration of LHRH antagonist therapy are selected such that efficient blockade of androgen secretion is obtained without the occurrence of the hormone surge that accompanies the use of LHRH agonists in other treatment methods. Preferably, the LHRH antagonist is administered to the subject prior to performing the procedure that removes or destroys prostatic tumor tissue. For example, an LHRH antagonist can be used in neoadjuvant hormonal downstaging therapy prior to radical prostatectomy (or other procedure to remove or destroy tumor tissue). Administration of an LHRH antagonist is preferably for a period sufficient to cause the prostate or prostatic tumor tissue to shrink in size prior to performing the COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-0815:5 11/ -7- 00 0 0 procedure that removes or destroys prostatic tumor tissue. A suitable period for preadministration of an LHRH antagonist typically is between about one month and Sabout one year. more preferably between about three months and about six months.

SUse of an LHRH antagonists in the combination treatment method is expected to 5 sufficiently reduce androgen production such that additional use of an antiandrogen is O not essential. However, in certain situations it may be desirable to use an antiandrogen and thus in another embodiment, this treatment method can further involve -administering an antiandrogen to the subject in combination with the LHRH antagonist v prior to performing the procedure that removes or destroys prostatic tumor tissue. Since use of LHRH antagonists avoid the agonist-induced hormone surge that occurs with SLHRH agonists, it is expected that when an antiandrogen is used in combination with 00 the LHRH antagonist, the dosage and duration of treatment with the antiandrogen would o be reduced as compared to when an antiandrogen is used in combination with an LHRH Cl agonist. In yet another embodiment, this treatment method can further involve administering one or more inhibitors of sex steroid biosynthesis to the subject in combination with the LHRH antagonist (optionally in further combination with an antiandrogen) prior to performing the procedure that removes or destroys prostatic tumor tissue.

Another aspect of the invention pertains to a method for treating prostate cancer in a subject in need of such treatment, comprising administering to the subject an LHRH antagonist; and simultaneously or subsequently administering to the subject an LHRH agonist- The type, dosage and duration of the combined LHRH antagonist and LHRH agonist therapy are selected such that the agonist-induced hormone surge is reduced or eliminated compared to when an LHRH agonist alone is used (discussed further in subsection II below). Thus, the LHRH agonist should be administered simultaneously with, or subsequent to, initiation of LHRH antagonist administration, but not before. In a preferred embodiment, an LHRH antagonist is administered to a subject for at least one week before an LHRH agonist is administered to the subject. Once LHRH agonist therapy has been initiated, the LHRH antagonist therapy can be continued the antagonist and the agonist can be coadministered) or the LHRH antagonist therapy can be discontinued first the LHRH antagonist alone is administered to the subject and then the LHRH agonist alone is administered to the subject). In a preferred embodiment, an LHRH antagonist and an LHRH agonist are coadministered for a period of one month to one year, more preferably for about three to six months. In certain embodiments, a procedure that removes or destroys tumor tissue a radical prostatectomy, cryosurgery or radiation therapy) is performed after the administration of the LHRH antagonist and LHRH agonist.

COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 0 12Z 40 -8- 00 0 As described above, while it may not be necessary to combine LHRH antagonist/LHRH agonist therapy with additional drugs, in certain situation it may be desirable to further combine the LHRH antagonist and LHRH agonist with other drugs, such as an antiandrogen and/or one or more inhibitors of sex steroid biosynthesis.

As is discussed in more detail below, a preferred route of administration for an LHRH antagonist (alone or in combination with an LHRH agonist) is by depot injection or other slow-release or sustained delivery method. A preferred route of antiandrogen administration is oral administration. Radical prostatectomy, cryosurgery or radiation therapy (external or interstitial) can be performed using standard methodologies.

The methods of the present invention can be applied to the treatment of prostate 00q cancer in male subjects at any stage of the cancer, although certain treatment methods 0 oare more preferred for particular cancer stages. For reviews on screening and diagnostic o methods for prostate cancer, see Gamick, M. (1993) Annals ofInternal Medicine 118:803-818; and Garnick, M. (1994) Scientific American 270:72-81. Prostate cancer is commonly evaluated according to a scale divided into four lettered stages: A, B, C and D. Tumors in stage A are microscopic; stage A 1 designates tumors confined to a relatively small area and composed of well-differentiated tissue, while stage A 2 tumors are more diffuse and less well differentiated. Stage B tumors are large enough to be felt during a rectal examination, while stage C prostate cancers have spread throughout the gland and typically have pushed past the borders of the prostate into surrounding structures. Stage D tumors have metastasized, to lymph nodes, bone, or other organs. Alternatively, tumors can be staged by the TNM staging system, in which tumors are ranked on a scale of progressively worsening disease from Tla to T4b T 1 c tumors are non-palpable and non-visible that were detected by elevated blood levels of prostate specific antigen). The methods of the invention are useful in the treatment of any stage of prostate cancer. However, it will be appreciated by the skilled artisan that methods involving procedures for removal or destruction of prostatic tumor tissue preferably are employed with non-metastasized cancers. For example, radical prostatectomy preferably is used with stage A, B and some stage C tumors where the tumor growth has not extended considerably beyond the borders of the prostate gland) as well as stage Tlc tumors. Radiation therapy external or interstitial) preferably is used with stage A. B or C tumors as well as Tic tumors.

To assess the efficacy of a treatment method of the invention, the size of the prostate can be determined by methods known in the art, for example, rectal examination, transrectal ultrasonography or magnetic resonance imaging (MRI).

Moreover, the size or extent of the prostate tumor (and metastatic tumors, if any) can. be assessed by known methods including a prostate-specific antigen blood test (describcJ further below), bone scanning, X-rays, skeletal survey, intravenous pyelography, CAT- COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-OE-O~sIB4# 13 1t40 -9- 00 o scan, MR, physical examination, biopsy, and the like. For treatment methods that involve surgery in neoadjuvant therapy wherein an LHRH antagonist is administered prior to a radical prostatectomy), the tumor can also be staged during surgery the prostate gland can be examined during surgery and/or a biopsy can be taken and examined). Thus, clinical staging and/or surgical staging may be used to oevaluate the extent of disease. Use of an LHRH antagonist in accordance with the methods of the invention is expected to result in a tumor stage, assessed at the time of _radical prostatectomy, that is improved compared to methodologies utilizing an LHRH agonist.

A preferred method of evaluating the extent of prostate cancer is to assay the Olevel of prostate-specific antigen (PSA) in a subject's blood. The PSA blood test is a 00 reasonably specific, sensitive, rapid, and inexpensive tool for screening for prostate cancer. In general, a blood PSA level above 4 ng/ml is considered to be suggestive of the presence of prostate cancer, with levels above 10 ng/ml being particularly indicative of cancer. For a subject undergoing treatment with an LHRH antagonist according to the methods of the invention, a pretreatment level of PSA can be established and the efficacy of the treatment assessed by monitoring periodically the PSA level in the subject's blood, wherein decreased PSA levels are used as an indicator of the efficacy of the treatment. The PSA nadir the point at which PSA levels do not decrease further even upon further treatment with an LHRH antagonist) can be used as the indicator point for initiation of a second therapy, for example for performance of a procedure that removes or destroys prostatic tumor tissue (such as radical prostatectomy, cryosurgery and/or radiation therapy). It is expected that the PSA nadir will be reached sooner using an LHRH antagonist, as compared to an LHRH agonist, since the hormone surge is avoided using an LHRH antagonist.

Additionally or alternatively, plasma concentrations of sex hormones can be monitored to assess the efficacy of the drug therapy. Concentrations of hormones such as testosterone, dihydrotestosterone, dehydroepiandrosterone (DHEA), DHEA-sulfate, androst-5-ene-3p, 17p-diol, and the estrogen 171-estradiol can all be measured by methods known the skilled artisan (see, F. Labrie et (1983) The Prosate 4:579).

Preferably, decreased levels of testosterone and dihydrotestosterone are used as indicators of treatment efficacy.

The response criteria for prostate developed by the National Prostate Cancer Project (see The Prostate, 1:375-382) can also be used to assess the efficacy of treatment. For treatment methods involving a procedure that removes or destroys tumor tissue (such as radical prostatectomy, cryosurgery, and/or radiation therapy), it is preferable to administer an LHI I antagonist until the size of the prostate or a prostate COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 4f 14/ 00 0 0 tumor has decreased and/or blood PSA levels have decreased before performing the C'-l procedure.

Although the methods of the invention are described in particular with application to the treatment of prostate cancer, it will be appreciated by the skilled artisan that these methods also can be applied to the treatment of other sex hormonedependent cancers, such as ovarian cancer or breast cancer, in humans or animals of either sex. In such cases, methods involving a step comprising surgical removal of tumor tissue are designed for the removal of the tumor tissue of the particular cancer to be treated.

C1 II. Methods for Inhibiting the Agonist-Induced Hormone Surge o Another aspect of the invention pertains to methods for inhibiting the agonistoinduced hormone surge that occurs due to LHRH agonist therapy, whatever the clinical setting treatment of a hormone-dependent cancer, such as prostate. ovarian or breast cancer with an LHRH agonist, treatment of endometriosis with an LHRH agonist, treatment of benign prostatic hypertropy with an LHRH agonist, and the like). As used herein, the term "agonist-induced hormone surge" "agonist-induced testosterone surge") is intended to refer to the transient increase in gonadotropic and gonadal steroid hormone secretion testosterone secretion) that results from LH agonist administration and that may last for several days in primates. As used herein, the term "inhibiting the agonist-induced hormone surge" is intended to refer to decreasing or diminishing the magnitude of gonadotropic and gonadal steroid hormone secretion. This inhibition may be partial but, more preferably, is a complete inhibition of the hormone surge. As demonstrated herein, the methods of the invention allow for complete elimination of the hormone surge. The methods generally involve administering to a subject in need of LHRH agonist therapy an LHRH antagonist in combination with the LHR.H agonist. In various embodiments, the methods of the invention involve the use of particular LHRH antagonist formulations, particular LHRH dosage regimens and/or a particular type of LHRH antagonist.

In one embodiment, the method of the invention for inhibiting agonist-induced hormone surge comprises: administering to the subject an LHRH antagonist in a sustained-release formulation; and administering to the subject an LHRH agonist; such that the agonist-induced hormone surge is inhibited in the subject In a preferred embodiment, the sustained-release formulation of LHRH antagonist is administered continuously for a period of at least about 14 days about 14 to 28 days. or one month) prior to administering the LHRH agonist. Administration COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:58 t 1 5/ -11- -ll- 00 O of the LHRH antagonist can be stopped prior to administering the LHRH agonist or.

alternatively, there can be a period of overlap in the LHRH antagonist treatment and the LHRH agonist treatment. In one embodiment, this overlap is for about one to three days, preferably one day. In another embodiment, this overlap is for about four to ten 5 days, preferably about seven days.

O As used herein, the term "sustained release formulation" is intended to encompass formulations that allow for the continuous delivery of an LHRH antagonist to a subject over a period of time, preferably several days to weeks. Such formulations are typically administered subcutaneously or intramuscularly and allow for the continual steady release of a predetermined amount of drug in the subject over time. The 0' sustained-release formulation of LHRH antagonist can be, for example, a formulation 00 comprising a polymer selected from the group consisting of a poly-lactide polymer, a o poly-glycolide polymer and a poly-lactide/poly-glycolide copolymer the drug is C encapsulated within a microcapsule comprising the polymer or copolymer). Such sustained-release formulations, suitable for depot injection, are known in the art for administration of LHRH agonists, such as leuprolide (see U.S. Patents 4,677,191 and 4,728,721; formulations are also described further in section III below). The sustained-release formulations can be formulated to allow for delivery of the drug over a predetermined time period. In another embodiment, the sustained-release formulation of LHRH antagonist comprises a formulaic in an osmotic pump the LHRH formulation is enclosed within the osmotic pump). Such osmotic pumps, which can be formulated to allow for release of a predetermined amount of drug over a predetermined time period, are known in the art the Alzet pump, commercially available from Alza, Palo Alto, CA). The dosage of LHRH antagonist released by the sustained-release formulation is preferably about 15-300 gg/kg/day, more preferably 15-200 pg/kg/day and even more preferably 15-100 pg/kg/day. As demonstrated in Example 4, complete elimination of the agonist-induced hormone surge has been achieved with LHRH antagonist pretreatment.

A preferred LHRH antagonist for administration in a sustained-release formulation has the following structure: Ac-D-Nall, 4-Cl-D-Phe 2 D-Pal 3 N-Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-AlaI 0 -LHRH (referred to herein as PPI-149). The LHRH agonist can be one of the many LHRH agonists known in the art, for example, leuprolide, goserelin. triptorelin, buserelin, nafarelin. lutrelin, cystorelin, gonadorelin or histrelin.

Preferred LHRH agonists are leuprolide, goserelin and triptorelin, most preferably leuprolide. In a preferred embodiment, the subject is a primate, most preferably a human.

In another embodiment, the method for inhibiting agonist-induced hormone surge comprises: COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08:15:56 16/ -12- 00 0 0 administering to the subject an LHRH antagonist for a period of at least 14 days at a dose of about 15-300 pg/kg/day; and administering to the subject an LHRH agonist; such that the agonist-induced hormone surge is inhibited in the subject.

C' 5 More preferably, the LHRH antagonist is administered at a dose of about 15-200 pg/kg/day and even more preferably at a dose of about 15-100 pg/kg/day. Preferably, the LHRH antagonist is administered continuously using a sustained-release formulation, encapsulated in a polymer microcapsule or an osmotic pump, as 0 discussed above).

LHRH antagonist administration can be stopped prior to administering the LHRH 00 agonist or, alternatively, there can be an overlap in LHRH antagonist and agonist Streatment an overlap of about one to three days, preferably one day, or an overlap o of about four to seven days, preferably about seven days). A preferred LHRH antagonist for use in the method is PPI-149. The LHRH agonist can be, for example, leuprolide, goserelin, triptorelin, buserelin, nafarelin, lutrelin, cystorelin, gonadorelin or histrelin.

Preferred LHRH agonists are leuprolide, goserelin and triptorelin, most preferably leuprolide. In a preferred embodiment, the subject is a primate, most preferably a human.

In yet another embodiment, the method for inhibiting agonist-induced hormone surge comprises: administering to the subject an LHRH antagonist having the following structure: Ac-D-Nall, 4-Cl-D-Phe 2 D-Pal 3 N-Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-Alal 0 LHRH; and administering to the subject an LHRH agonist; such that the agonist-induced hormone surge is inhibited in the subject.

Preferably, the LHRH antagonist is administered for.a period of at least 14 days.

Preferably, the dosage of LHRH antagonist is about 15-300 pg/kg/day, more preferably about 15-200 pg/kg/day, and even more preferably about 15-100 pg/kg/day. Preferably, the LHRH antagonist is administered continuously using a sustained-release formulation, encapsulated in a polymer microcapsule or an osmotic pump, as discussed above).

LHRH antagonist administration can be stopped prior to administering the LHRH agonist or, alternatively, there can be an overlap in LHRH antagonist and agonist treatment an overlap of about one to three days, preferably one day, or an overlap of about four to seven days, preferably about seven days). The LHRH agonist can be, for example, leuprolide, goserelin, triptorelin, buserelin. nafarelin, lutrelin, cystorelin, gonadorelin or histrelin. Preferred LHRH agonists are leuprolide, goserelin and COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08:15:56 17/ -13- O 00 0 triptorelin. most preferably leuprolide. In a preferred embodiment, the subject is a primate, most preferably a human.

In an especially preferred embodiment, the method for inhibiting agonist-induced hormone surge comprises: C 5 administering to the subject an LHRH antagonist having the following O structure: Ac-D-Nal 1 4-Cl-D-Phe 2 D-Pal 3 N-Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-Ala 0.

LHRH at a dosage of about 15-300 ig/kg/day in a sustained-release formulation for a period of at least about 14 days; and Qadministering to the subject an LHRH agonist; such that the agonist-induced hormone surge is inhibited in the subject.

0< More preferably, the dosage of LHRH antagonist is about 15-200 gg/kg/day and 0 0 even more preferably about 15-100 pg/kg/day. Preferably, the sustained-release 0 formulation is a formulation comprising a polymer selected from the group consisting of l a poly-lactide polymer, a poly-glycolide polymer and a poly-lactide/poly-glycolide copolymer or comprises a formulaic in an osmotic pump. Administration of the LHRH antagonist can be stopped prior to administering the LHRH agonist or, alternatively, there can be an overlap in LHRH antagonist and agonist treatment an overlap of about one to three days, preferably one day, or an overlap of about four to seven days, preferably about seven days). The LHRH agonist can be, for example, leuprolide, goserelin, triptorelin, buserelin, nafarelin, lutrelin, cystorelin, gonadorelin or histrelin.

Preferred LHRH agonists are leuprolide, goserelin and triptorelin, most preferably leuprolide. In a preferred embodiment, the subject is a primate, most preferably a human.

In a particular subembodiment of the methods for inhibiting agonist-induced hormone surge, the invention provides a method for inhibiting the "acute-on-chronic" hormone surge phenomenon. The "acute-on-chronic" hormone surge phenomenon refers to the periodic increases in hormone levels that can occur with each readministration of an LHRH agonist to a subject undergoing LHRH agonist therapy, after the initial hormone surge that accompanied the initial administration of LHRH agonist has occurred. The invention provides a method for inhibiting "acute-on-chronic" agonist-induced hormone surge caused by LHRH agonist therapy in a subject in need of LHRH agonist therapy, comprising: administering to the subject an LHRH agonist at regular intervals; and co-administering to the subject an LHRH antagonist with the LHRH agonist during each interval; such that the "acute-on-chronic" agonist-induced hormone surge is inhibited in the subject.

COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 18/ 14- 00 0 The regular intervals at which the LHRH agonist is administered can be. for example, monthly or weekly intervals. According to the method of the invention, the LHRH antagonist is coadministered to the subject during these intervals monthly or weekly). Coadministration of the LHRH antagonist "during each interval" is intended C 5 to include administering the LHRH antagonist to the subject concurrently with the LHRH agonist the LHRH agonist and LHRH antagonist each may be administered as a single monthly or weekly injection, each administered at the same time). Moreover, n coadministration of the LHRH antagonist "during each interval" is intended to include O administering the LHRH antagonist to the subject for a period before the LHRH agonist readministration occurs, and possibly continuing for period after the LHRH agonist ,00 readministration has ceased. For example, suppose the LHRH agonist readministration O comprises a single injection, once monthly. To inhibit the "acute-on-chronic" agonisto induced hormone surge, LHRH antagonist administration can be begun for a short period one to five days) prior to the single LHRH agonist injection, and LHRH antagonist administration can be continued through the day of the LHRH agonist injection and, if necessary, continued for a short period one to five days) after the LHRH agonist injection. To continually administer LHRH antagonist to the subject during this period, daily injections of the LHRH antagonist can be performed or, alternatively, slow-release formulation of the LHRH antagonist, that provides for continuous release of LHRH antagonist in the subject over the course of this period can be used.

Preferably, the LHRH antagonist is co-administered at a dosage of about 15-300 Pg/kg/day, more preferably at a dosage of about 15-200 pg/kg/day, and even more preferably at a dosage of about 15-100 pg/kg/day. A preferred LHRH antagonist for use in the method has the following structure: Ac-D-Nall, 4-Cl-D-Phe 2 D-Pal3, N-Me- Tyr 5 D-Asn 6 Lys(iPr) 8 D-Ala t l-LHRH. Preferred LHRH agonist include leuprolide, goserelin, triptorelin, buserelin, nafarelin, lutrelin, cystorelin, gonadorelin and histrelin (most preferably, leuprolide). In a preferred embodiment, the subject is a primate (preferably, a human).

III. Pharmaceutical Compositions LHRH antagonists suitable for use in the methods of the invention can be incorporated into pharmaceutical compositions suitable for administration to a subject.

Preferably, an LHRH antagonist alone is formulated into the pharmaceutical composition, although in certain embodiments the LHRH antagonist may be combined with one or more other drugs such as an LHRH agonist. antiandrogen and/or inhibitor of sex steroid biosynthesis (collectively referred to as "combination drug(s)"). In a COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 19/ 00 0 O preferred embodiment, the pharmaceutical composition comprises an LHRH antagonist and a pharmaceutically acceptable carrier.

A "therapeutically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired result. A therapeutically effective amount of an LHRH antagonist may vary according to factors such as the Odisease state, age, and weight of the individual, and the ability of the LHRH antagonist (alone or in combination with one or more combination drugs) to elicit a desired response in the individual. Dosage regimens may be adjusted to provide the optimum therapeutic response. A therapeutically effective amount is also one in which any toxic or detrimental effects of the antagonist are outweighed by the therapeutically beneficial 0effects. A non-limiting range for a therapeutically effective amount of an LHRH 00 antagonist is 0.01 gg/kg- 10 mg/kg, preferably between about 0.01 and 5 mg/kg. It is to obe noted that dosage values may vary with the severity of the condition to be alleviated.

C It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that dosage ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition. For suppression of agonist-induced hormone surge, preferred dosages of LHRH antagonist are as described above in section I.

As used herein, "pharnmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. Preferably, the carrier is suitable for intravenous, intramuscular, subcutaneous or parenteral administration by injection). Depending on the route of administration, the active compound may be coated in a material to protect the compound from the action of acids and other natural conditions which may inactivate the compound.

An LHRH antagonist can be administered by a variety of methods known in the art. In a preferred embodiment, the LHRH antagonist is administered in a time release formulation (also referred to as a sustained-release formulation), for example in a composition which includes a slow release polymer, or a composition suitable for depot injection. The active compounds can be prepared with carriers that will protect the compound against rapid release, such as a controlled release formulation, including implants. transdermal patches, and microencapsulated delivery systems. Biodegradable.

biocormpatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen,. polyorthoesters, polylactic acid and polylactic, polyglycolic copolymers (PLG). Many methods for the preparation of such formulations are patented or generally known to those skilled in the art. See, Sustained and Controlled Release COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08:15:56 2 20/ -16- 00 0 O Drug Delivery Systems. J.R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.

Particularly preferred formulations include controlled-release compositions such as are known in the art for the administration of leuprolide (trade name: Lupron!), e.g., microcapsules Patents 4.652,441 and 4,917,893), injectable formulations (U.S.

C 5 Patent 4.849,228), lactic acid-glycolic acid copolymers useful in making microcapsules or injectable formulations Patents 4,677,191 and 4,728,721), and sustained-release compositions for water-soluble polypeptides Patent 4,675,189).

When appropriately formulated, an LHRH antagonist may be orally Oadministered, for example, with an inert diluent or an assimilable edible carrier. The LHRH antagonist (and other ingredients may also be enclosed in a hard or soft shell 00 gelatin capsule, compressed into tablets, or incorporated directly into the subject's diet.

oFor oral therapeutic administration, the LHRH antagonist may be incorporated with Oexcipients and used in the form of ingestible tablets, buccal tablets, troches, capsules.

elixirs. suspensions, syrups, wafers, and the like. The percentage of the LHRI-I antagonist in the compositions and preparations may, of course, be varied. The amount of the LHRH antagonist in such therapeutically useful compositions is such that a suitable dosage will be obtained.

To administer an LIRH antagonist (alone or with one or more combination drugs) by other than parenteral administration, it may be necessary to coat the compound with, or co-administer the compound with, a material to prevent its inactivation. For example. the LHRII antagonist may be administered to a subject in an appropriate carrier, for example, liposomes, or a diluent. Pharmaceutically acceptable diluents include saline and aqueous buffer solutions. Liposomes include water-in-oil-in-water COF emulsions as well as conventional liposomes (Strejan el al, (1984) Neuroimmunol. 7:27). Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the pharmaceutical compositions of the invention is r-,ntemplated. Supplementary active compounds can also be incorporated into the t ipositions.

Therapeutic compositions typically must be s Ie and stable under the conditions of manufacture and storage. The composition can be urmulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration.

The carrier can be a solvent or dispersion medium containing, for example, water. ethanol.

polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol. and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-03; 15:556 1/4 Al 21 17- 00 O the case of dispersion and by the use of surfactants. in many cases, it will be preferable to (N include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays -absorption, for example, monostearate salts and gelatin.

O Sterile injectable solutions can be prepared by incorporating the active compound LJ-IZI- antagonist) in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other 0 ingredients from those enumerated above. In the case of sterile powders for the 00 preparation of sterile injectable solutions, the preferred methods of preparation are o vacuum drying and freeze-drying which yields a powder of the active ingredient plus (N any additional desired ingredient from a previously sterile-filtered solution thereof.

Dosage regimens may be adjusted to proi/ide the optimum therapeutic response.

For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is especially advantageous to formulate parenteral compositions in dosage unit formn for ease of administration and uniformity of' dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the mammalian subjects 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, and the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in individuals.

This invention is further illustrated by the following examples which should not be construed as limiting. The contents of all references, patents and published patent applications cited throughout this application are hereby incorporated by reference.

Example I A human subject with a localized prostate tumor Stage B) is treated according to a method of the invention as follows: An LHRH antagonist having low histamine-releasing activity is administered in a depot formulation by intramuscular injection of 10 mg of the LHRH antagonist in lactic acid/glycolic acid copolymer microcapsules (microcapsules are prepared as COMS ID No: ARCS-i189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 22/ -18- 00 O described in U.S. Patent 4,849,228) or, alternatively, using a suitable pump for continuous drug delivery. Additional depot injections are administered each month.

typically for a total treatment period of three to six months (although longer treatment periods can be used according to the individual need and the professional judgment of Cl 5 the person supervising the therapy). The size and progression of the prostate tumor is monitored by transrectal ultrasonography, rectal examination, and assay for prostatespecific antigen. A radical prostatectomy, cryosurgery or.radiation therapy (external or Sinterstitial) is performed by standard techniques when the prostate tumor has decreased Sin size or extent and/or the PSA nadir has been reached. LHRH antagonist administration can be continued following radical prostatectomy, cryosurgery or Cl radiation therapy according to the individual need and the professional judgment of the o person supervising the therapy.

Example 2 A human subject with a localized Stage B) prostate cancer is treated with an LHRH antagonist as described in Example 1. After one month of treatment with the LHRH antagonist, an LHRH agonist leuprolide, goserelin or triptorelin) is administered. For example, leuprolide is administered in a depot formulation by intramuscular injection of 7.5 mg of leuprolide in lactic acid/glycolic acid copolymer microcapsules (microcapsules are prepared as described in U.S. Patent 4,849,228). Oral flutamide (250 mg every 8 hours) is also begun. Treatment with the LHRH antagonist is continued such that the subject is treated with the LHRH antagonist, leuprolide and flutamide in combination. The progression of the prostate tumor is monitored as described in Example 1. If necessary, after a period of treatment sufficient to reduce the size and progression of the prostate tumor or reach the PSA nadir three to six months), a radical prostatectomy, cryosurgery or radiation therapy (external or interstitial) is performed by standard techniques.

Example 3 A human subject with metastasized Stage D) prostate cancer is treated with an LHRH antagonist as described in Example 2 for one month after which time LHRH antagonist therapy is discontinued and the subject is further treated with a combination of an LHRH agonist leuprolide, goserelin or triptorelin) and an antiandrogen flutamide), also as described in Example 2, thus ameliorating the agonist-induced testosterone surge expected to be produced by use of an LHRH agonist.

COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 23/ -19- 00 SExample 4 In this example, plasma testosterone levels were measured in adult male rats or cynomolgus monkeys treated with a combination of an LHRH antagonist and an LHRH agonist to determine whether prior treatment with an LHRH antagonist could prevent the agonist-induced testosterone surge normally induced by administration of the LHRH O agonist.

In a first series of experiments, adult male rats were first administered an LHRH antagonist having the structure: Ac-D-Nal 4-Cl-D-Phe 2 D-Pal 3 N-Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-Ala 0 O LH

RH

00 S(referred to herein as PPI-149). PPI-149 was administered via a subcutaneous Ci continuous osmotic minipump the Alzet pump, commercially available from Alza, Palo Alto, CA) at a dose of 15 pg/kg/day or 50 pg/kg/day. PPI-149 administration continued for a total of 15-28 days according to several different protocols. A second pump releasing leuprolide (trade name: Lupron®) at 1.8 pg/kg/day was implanted such that there was an overlap ofO, 1 or 7 days with the previously-implanted pump releasing PPI-149. Results from several independent experiments are shown in Figures 1-4.

In the experiment shown in Figure 1, PPI-149 (at 15 pg/kg/day or 50 pg/kg/day) was administered by osmotic pump for 28 days and then this pump was removed and a second pump administering leuprolide (at 1.8 pg/kg/day) was implanted such that there was no overlap between the antagonist and agonist treatment. Plasma testosterone levels (in ng/ml) were measured during the prolonged administration of PPI-149 alone and following replacement of the PPI-149 pump with the leuprolide pump. The results shown in the graph of Figure 1 demonstrate that PPI-149 at 50 pLg/kg/day suppressed plasma testosterone levels to castrate levels and prevented the leuprolide-induced testosterone surge when administered prior to agonist treatment and without any overlap between the antagonist and agonist treatment.

In the experiment shown in Figure 2, PPI-149 (at 50 gg/kg/day) was administered by osmotic pump for 15 days and then a second pump administering leuprolide (at 1.8 gg/kg/day) was implanted such that there was one day of overlap between the antagonist and agonist treatment the first PPI-149-releasing pump was removed one day after implantation of the second leuprolide-releasing pump). Plasma testosterone levels were measured during the prolonged administration of PPI-149 alone and following removal of the PPI-149 pump when leuprolide alone was administered.

The results shown in the graph of Figure 2 demonstrate that PPI-149 at 50 pg/kg/day suppressed plasma testosterone levels to castrate levels and prevented the leuprolide- COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 24/ 00 0 induced testosterone surge when administered prior to agonist treatment and with one .day of overlap between the antagonist and agonist treatment.

In the experiment shown in Figure 3, PPI-149 (at 15 tg/kg/day or 50 g/kg/day) was administered by osmotic pump for 28 days and then a second pump administering C' 5 leuprolide (at 1.8 Ag/kg/day) was implanted such that there was one day of overlap between the antagonist and agonist treatment the first PPI-149-releasing pump was removed one day after implantation of the second leuprolide-releasing pump). Plasma CT testosterone levels were measured during the prolonged administration of PPI-149 alone 0and following removal of the PPI-149 pump when leuprolide alone was administered.

The results shown in the graph of Figure 3 demonstrate that PPI-149 at 50 pg/kg/day 0 suppressed plasma testosterone levels to castrate levels and prevented the leuprolideo induced testosterone surge when administered prior to agonist treatment and with one o day of overlap between the antagonist and agonist treatment.

In the experiment shown in Figure 4, PPI-149 (at 15 jg/kg/day or 50 lg/kg/day) was administered by osmotic pump for 28 days and then a second pump administering leuprolide (at 1.8 lg/kg/day) was implanted such that there were seven days of overlap between the antagonist and agonist treatment the first PPI-149-releasing pump was removed seven days afer implantation of the second leuprolide-releasing pump).

Plasma testosterone levels were measured during the prolonged administration of PP1- 149 alone and following removal of the PPI-149 pump when leuprolide alone was administered. The results shown in the graph of Figure 4 demonstrate that PPI-149 at jg/kg/day suppressed plasma testosterone levels to castrate levels and prevented the leuprolide-induced testosterone surge when administered prior to agonist treatment and with seven days of overlap between the antagonist and agonist treatment.

Thus, in adult male rats PPI-149 at 50 pg/kg/day can prevent the testosterone surge induced by LHRH agonist administration when PPI-149 is administered either 0, 1 or 7 days prior to administration of the LHRH agonist.

In a second series of experiments, male cynomolgus monkeys were administered the LHRH antagonist PPI-149 via a subcutaneous continuous osmotic minipump (Alzet pump; Alza, Palo Alto, CA) at a dose of 0, 30, 100 or 300 lg/kg/day for a period of 28 days. On day 28, pumps were removed and monkeys were administered a single intramuscular dose of the depot, sustained-release (one-month) formulation of Lupron®, equivalent to 0.16 mg/kg. This dose of leuprolide is 1.6 times that used in the treatment of prostate cancer in humans (0.1 mg/kg) and 3.2 times that used in the treatment of endometriosis (0.05 mg/kg).

The results of these experiments, shown graphically in Figure 5, demonstrate that a dose of 100 gg/kg/day of PPI-149 induces a rapid, complete medical castration within COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 25/ -21 00 o the first two days of treatment. In addition, the same dose ofPPI-149 is able to induce a C' .complete blockade of the testosterone surge that typically results from the depot formulation of Lupron®. Since a high dose of leuprolide was used in these experiments 1.6 to 3.2 times the dose used clinically), it is expected that the same treatment C 5 protocol with PPI-149 will block the agonist-induced testosterone surge induced by the o three-month depot formulation of Lupron®, as well as the one- and three-month formulations of Zoladex® (gosarelin) and other LHRH agonists. such as triptorelin (Decapeptyl; D-Trp-6-LHRH), that are currently on the market.

SEQUIVALENTS

00 Those skilled in the art will recognize, or be able to ascertain using no more than Sroutine experimentation, many equivalents to the specific embodiments of the invention Cl described herein. Such equivalents are intended to be encompassed by the following claims.

COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02

Claims (61)

1. 2. The method of claim 1, wherein the LHRH antagonist is administered to the subject prior to performing the procedure that removes or destroys prostatic tumor Clq tissue. 00
2. 3. The method of claim 2, wherein the LHRH antagonist is administered for about 3 to 6 months prior to performing the procedure that removes or destroys prostatic tumor tissue.
3. 4. The method of claim 2, wherein the LHRH antagonist is administered for about 6 to 12 months prior to performing the procedure that removes or destroys prostatic tumor tissue. The method of claim 2, wherein administration of the LHRH antagonist to the subject is continued after performing the procedure that removes or destroys prostatic tumor tissue.
4. 6. The method of claim 1, wherein the procedure that removes or destroys prostatic tumor tissue is selected from the group consisting of radical prostatectomy, cryosurgery, external radiation therapy and interstitial radiation therapy.
5. 7. The method of claim 1, wherein the LHRH antagonist has an EDso for histamine release in a standard in vitro histamine release assay of at least 3 gg/ml.
6. 8. The method of claim 1, wherein the LHRH antagonist has an ED 5 0 for histamine release in a standard in vitro histamine release assay of at least 5 gg/ml.
7. 9. The method of claim 1, wherein the LHRH antagonist has an ED 5 0 for histamine release in a standard in vitro histamine release assay of at least 10 ptg/m 1 COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 V- 27/ -23- 00 0 O 10. The method of claim I, wherein the LHRH antagonist has the following structure: Ac-D-Nall. 4-CI-D-Phe 2 D-Pal 3 N-Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-Ala 10 t LHRH. ci 5 11. The method of claim 2, further comprising administering to the subject an 0 antiandrogen in combination with the LHRH antagonist prior to performing the procedure that removes or destroys prostatic tumor tissue.
8. 12. The method of claim 11, wherein the antiandrogen is a steroidal antiandrogen. 0 O 13. The method of claim 11, wherein the antiandrogen is a nonsteroidal o antiandrogen.
9. 14. The method of claim 11, wherein the antiandrogen is selected from the group consisting of flutamide, bicalutamide and nilutamide. The method of claim 2, further comprising administering to the subject at least one inhibitor of sex steroid biosynthesis in combination with the LHRH antagonist prior to performing the procedure that removes or destroys prostatic tumor tissue.
10. 16. A method for treating prostate cancer in a subject in need of such treatment, comprising: administering to the subject an LHRH antagonist; and simultaneously or subsequently administering to the subject an LHRH agonist.
11. 17. The method of claim 16, wherein the LHRH agonist is administered to the subject subsequent to administration of the LHRH antagonist to the subject.
12. 18. The method of claim 16, wherein the LHRH agonist is administered to the subject simultaneously with administration of the LHRH antagonist to the subject.
13. 19. The method of claim 16, wherein the LHRH agonist is leuprolide. The method of claim 16, wherein the LHRH agonist is selected from the group consisting ofgoserelin, triptorelin, buserelin, nafarelin, lutrelin, cystorelin, gonadorelin and histrelin. COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:58 28/
14. 24- 00 0 0 21. The method of claim 16, wherein the LHRH antagonist has an ED 50 for 4 histamine release in a standard in virro histamine release assay of at least 3 Ag/ml. C 5 22. The method of claim 16, wherein the LHRH antagonist has an ED 5 0 for histamine release in a standard in vitro histamine release assay of at least 5 Jig/ml. 23. The method of claim 16, wherein the LHRH antagonist has an ED 50 for histamine release in a standard in vitro histamine release assay of at least 10 pg/ml. C 24. The method of claim 16, wherein the LHRH antagonist has the following 0 o structure: Ac-D-Na 1 4-Cl-D-Phe 2 D-Pal 3 N-Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-Ala t 0 LHRH.
15. 25. The method of claim 16, further comprising administering to the subject an antiandrogen.
16. 26. The method of claim 25, wherein the antiandrogen is a steroidal antiandrogen.
17. 27. The method of claim 25, wherein the antiandrogen is a nonsteroidal antiandrogen.
18. 28. The method of claim 25, wherein the antiandrogen is selected from the group consisting offlutamide, bicalutamide and nilutamide.
19. 29. The method of claim 16, further comprising performing a procedure on the subject that removes or destroys prostatic tumor tissue subsequent to administration of the LHRIH antagonist and the LHRH agonist to the subject. The method of claim 29, wherein the procedure that removes or destroys prostatic tumor tissue is selected from the group consisting of radical prostatectomy, cryosurgery, external radiation therapy and interstitial radiation therapy.
20. 31. A method for inhibiting agonist-induced hormone surge caused by LHRH agonist therapy in a subject in need of LHRH agonist therapy, comprising: administering to the subject an LHRH antagonist in a sustained-release formulation; and COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 29/ 25 00 0 administering to the subject an LHRH agonist; such that the agonist-induced hormone surge is inhibited in the subject.
21. 32. The method of claim 31, wherein the LHRH antagonist is administered continuously for a period of at least about 14 days prior to administering the LHRH Sagonist.
22. 33. The method of claim 31, wherein LHRH antagonist administration is Cf stopped prior to administering the LHRH agonist. o 34. The method of claim 31, wherein LHRH antagonist administration 00 overlaps for about one to three day with LHRH agonist administration. 0 Cl 35. The method of claim 31, wherein LHRH antagonist administration overlaps for about four to ten days with LHRH agonist administration.
23. 36. The method of claim 31, wherein the sustained-release formulation of LHRH antagonist is a formulation comprising a polymer selected from the group consisting of a poly-lactide polymer, a poly-glycolide polymer and a poly-lactide/poly- glycolide copolymer.
24. 37. The method of claim 31, wherein the sustained-release formulation of LHRH antagonist comprises a formulaic in an osmotic pump.
25. 38. The method of claim 31, wherein the dosage of LHRH antagonist is about 15-300 lg/kg/day.
26. 39. The method of claim 38, wherein the dosage of LHRH antagonist is about 15-200 tg/kg/day. The method of claim 39, wherein the dosage of LHRH antagonist is about 15-100 pg/kg/day.
27. 41. The method of claim 31, wherein the LHRH antagonist has the following structure: Ac-D-Nall, 4-Cl-D-Phe 2 D-Pal 3 N-Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-AlalO- LHRH.
28. 42. The method of claim 31, wherein the LHRH agonist is leuprolide. COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 30/ 26- 00 0 0
29. 43. The method of claim 31, wherein the LHRH agonist is selected from the group consisting of goserelin, triptorelin, buserelin, nafarelin, lutrelin. cystorelin. gonadorelin and histrelin. O 44. The method of claim 31, wherein the subject is a primate. A method for inhibiting agonist-induced hormone surge caused by LHRH Sagonist therapy in a subject in need of LHRH agonist therapy, comprising: administering to the subject an LHRH antagonist for a period of at least 0 N about 14 days at a dose of about 15-300 gg/kg/day; and 00 administering to the subject an LHRH agonist; O O such that the agonist-induced hormone surge is inhibited in the subject.
30. 46. The method of claim 45, wherein the LHRH antagonist is administered at a dose of about 15-200 pg/kg/day.
31. 47. The method of claim 45, wherein the LHRH antagonist is administered at a dose of about 15-100 pg/kg/day.
32. 48. The method of claim 45, wherein the LHRH antagonist is administered continuously using a sustained-release formulation.
33. 49. The method of claim 48, wherein the sustained-release formulation of LHRH antagonist is a formulation comprising a polymer selected from the group consisting of a poly-lactide polymer, a poly-glycolide polymer and a poly-lactide/poly- glycolide copolymer. The method of claim 48, wherein the sustained-release formulation of LHRH antagonist comprises a formulaic in an osmotic pump.
34. 51. The method of claim 45, wherein LHRH antagonist administration is stopped prior to administering the LHRH agonist.
35. 52. The method of claim 45, wherein LHRH antagonist administration overlaps for about one to three day with LHRH agonist administration. COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05--08;15:56 31/ -27- 00 O0 O 53. The method of claim 45, wherein LHRH antagonist administration l overlaps for about four to ten days with LHRH agonist administration.
36. 54. The method of claim 45, wherein the LHRH antagonist has the following structure: Ac-D-Nall, 4-CI-D-Phe 2 D-Pal 3 N-Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-Ala 10 SLHRH. The method of claim 45, wherein the LHRH agonist is leuprolide.
37. 56. The method of claim 45, wherein the LHRH agonist is selected from the 0 group consisting of goserelin, triptorelin, buserelin, nafarelin, lutrelin, cystorelin, 00 gonadorelin and histrelin. 0 C 57. The method of claim 45, wherein the subject is a primate.
38. 58. A method for inhibiting agonist-induced hormone surge caused by LHRH agonist therapy in a subject in need of LHRH agonist therapy, comprising: administering to the subject an LHRH antagonist having the following structure: Ac-D-Nall, 4-CI-D-Phe 2 D-Pal 3 N-Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-Alal 0 LHRH; and administering to the subject an LHRH agonist; such that the agonist-induced hormone surge is inhibited in the subject.
39. 59. The method of claim 58, wherein the LHRH antagonist is administered for a period of at least about 14 days. The method of claim 58, wherein the dosage of LHRH antagonist is about 15-300 pg/kg/day.
40. 61. The method of claim 60, wherein the dosage of LHRH antagonist is about 15-200 gg/kg/day.
41. 62. The method of claim 61, wherein the dosage of LHRH antagonist is about 15-100 pg/kg/day.
42. 63. The method of claim 58, wherein the LHRH antagonist is administered continuously using a sustained-release formulation. COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15: 56 32/ -28- 00 O
43. 64. The method of claim 63, wherein the sustained-release formulation of c LHRH antagonist is a formulation comprising a polymer selected from the group consisting of a poly-lactide polymer. a poly-glycolide polymer and a poly-lactide/poly- glycolide copolymer. (CM The method of claim 63, wherein the sustained-release formulation of LHRH antagonist comprises a formulaic in an osmotic pump. n
44. 66. The method of claim 58, wherein LHRH antagonist administration is stopped prior to administering the LHRH agonist. 0 00 67. The method of claim 58, wherein LHRH antagonist administration O overlaps for about one to three day with LHRH agonist administration.
45. 68. The method of claim 58, wherein LHRH antagonist administration overlaps for about four to ten days with LHRH agonist administration.
46. 69. The method of claim 58, wherein the LHRH agonist is leuprolide.
47. 70. The method of claim 58, wherein the LHRH agonist is selected from the group consisting of goserelin, triptorelin, buserelin, nafarelin, lutrelin, cystorelin, gonadorelin and histrelin.
48. 71. The method of claim 58, wherein the subject is a primate.
49. 72. A method for inhibiting agonist-induced hormone surge caused by LHRH agonist therapy in a subject in need of LHRH agonist therapy, comprising: administering to the subject an LHRH antagonist having the following structure: Ac-D-Nall, 4-Cl-D-Phe 2 D-Pal 3 N-Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-Ala 0 O- LHRH at a dosage of about 15-300 fg/kg/day in a sustained-release formulation for a period of at least about 14 days; and administering to the subject an LHRH agonist; such that the agonist-induced hormone surge is inhibited in the subject.
50. 73. The method of claim 72, wherein the dosage of LHRH antagonist is about 15-200 pg/kg/day. COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 4 33/ -29- 00 O0 o 74. The method of claim 73, wherein the dosage of LHRH antagonist is CI about 15-100 pg/kg/day. The method of claim 72, wherein the sustained-release formulation of LHRH antagonist is a formulation comprising a polymer selected from the group Sconsisting of a poly-lactide polymer, a poly-glycolide polymer and a poly-lactide/poly- glycolide copolymer. C 76. The method of claim 72, wherein the sustained-release formulation of O 10 LHRH antagonist comprises a formulaic in an osmotic pump. 0 00 77. The method of claim 72, wherein LHRH antagonist administration is Sstopped prior to administering the LHRH agonist.
51. 78. The method of claim 72, wherein LHRH antagonist administration overlaps for about one to three day with LHRH agonist administration.
52. 79. The method of claim 72, wherein LHRH antagonist administration overlaps for about four to ten days with LHRH agonist administration. The method of claim 72, wherein the LHRH agonist is leuprolide.
53. 81. The method of claim 72, wherein the LHRH agonist is selected from the group consisting ofgoserelin, triptorelin, buserelin, nafarelin, lutrelin, cystorelin. gonadorelin and histrelin.
54. 82. The method of claim 72, wherein the subject is a primate.
55. 83. A method for inhibiting "acute-on-chronic" agonist-induced hormone surge caused by LHRH agonist therapy in a subject in need of LHRH agonist therapy, comprising: administering to the subject an LHRH agonist at regular intervals: and co-administering to the subject an LHRH antagonist with the LHRH agonist during each interval; such that the "acute-on-chronic" agonist-induced hormone surge is inhibited in the subject.
56. 84. The method of claim 83, wherein the regular intervals are monthly. COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02 02-05-08;15:56 34/ 00 0 0 Cl 85. The method of claim 83, wherein the regular intervals are weekly.
57. 86. The method of claim 83, wherein the LHRH antagonist is co- administered at a dosage of about 15-300 pg/kg/day.
58. 87. The method of claim 86, wherein the LHRH antagonist is co- _administered at a dosage of about 15-200 pg/kg/day.
59. 88. The method of claim 87, wherein the LHRH antagonist is co- o administered at a dosage of about 15-100 jg/kg/day. 00 S89. The method of claim 83, wherein the LHRH antagonist has the following CN structure: Ac-D-Nall, 4-C1-D-Phe 2 D-Pal 3 N-Me-Tyr 5 D-Asn 6 Lys(iPr) 8 D-Ala 0- LHRH. The method of claim 83, wherein the LHRH agonist is leuprolide.
60. 91. The method of claim 83, wherein the LHRH agonist is selected from the group consisting of goserelin, triptorelin, buserelin, nafarelin, lutrelin, cystorelin, gonadorelin and histrelin.
61. 92. The method of claim 83, wherein the subject is a primate. COMS ID No: ARCS-189095 Received by IP Australia: Time 16:11 Date 2008-05-02