AU708475B2

AU708475B2 - Conjugates useful in the treatment of benign prostatic hyperplasia

Info

Publication number: AU708475B2
Application number: AU74321/96A
Authority: AU
Inventors: Deborah Defeo-Jones; Victor M. Garsky; Raymond E Jones; Allen I. Oliff; Edward M. Scolnick
Original assignee: Merck and Co Inc
Current assignee: Merck and Co Inc
Priority date: 1995-10-18
Filing date: 1996-10-15
Publication date: 1999-08-05
Anticipated expiration: 2016-10-15
Also published as: JP2000506494A; AU7432196A; EP0855910A4; EP0855910A1; WO1997014416A1

Description

WO 97/14416 PCT/US96/1649 0 -1- TITLE OF THE INVENTION CONJUGATES USEFUL IN THE TREATMENT OF BENIGN PROSTATIC

HYPERPLASIA

BACKGROUND OF THE INVENTION Benign prostate hyperplasia (or "prostatism") can be seen in almost 100 percent of all men over the age of 80, and changes in the prostate can be discovered in about 50 percent of men by the time they reach the age of 60. Many men with benign prostate hyperplasia

(BPH)

remain without symptoms, others show slow progression, while others remain stable. However, some 400,000 men a year have symptoms severe enough to require surgery. The most common surgery, transurethral resection, is effective in relieving the symptoms of BPH, although side-effects, including morbidity from the operation itself, mild to severe urinary incontinence and some degree of erectile or ejaculatory dysfunction, have been reported in a limited number of patients.

Normally the prostate remains stable until after the age of when the tissue begins to change, growing and causing the size of the prostate to increase. The enlarging prostate squeezes the urethra, producing the symptoms that characterize BPH. These include difficulty in starting urination (hesitancy), a weak urinary stream, dribbling after urination, and increased frequency or urgency to urinate during the sleep period. Sometimes urination may be painful. The symptoms of obstruction of the urethra can often become more severe if a urinary infection develops, one of the common complications of BPH.

Prostate specific Antigen (PSA) is a single chain 33 kDa glycoprotein that is produced almost exclusively by the human prostate epithelium and occurs at levels of 0.5 to 2.0 mg/ml in human seminal fluid (Nadji, Taber, Castro, et al. (1981) Cancer 4 8 :1 2 29;Papsidero, Kuriyama, Wang, et al. (1981).

JNCI

66:37; Qui, Young, Bihartz, et al. (1990), J. Urol.

144:1550; Wang, Valenzuela, Murphy, et al. (1979).

Invest. Urol. 17:159). The single carbohydrate unit is attached at asparagine residue number 45 and accounts for 2 to 3 kDa of the total WO 97/14416 PCT/US96/1 6 4 9 0 -2molecular mass. PSA is a protease with chymotrypsin-like specificity (Christensson, Laurell, Lilja, H. (1990). Eur. J. Biochem.

194:755-763). It has been shown that PSA is mainly responsible for dissolution of the gel structure formed at ejaculation by proteolysis of the major proteins in the sperm entrapping gel, Semenogelin I and Semenogelin II, and fibronectin (Lilja, H. (1985). J. Clin. Invest.

76:1899; Lilja, Oldbring, Rannevik, et al. (1987) J. Clin.

Invest. 80:281; McGee, Herr, J.C. (1988). Biol. Reprod. 39:499).

The PSA mediated proteolysis of the gel-forming proteins generates several soluble Semenogelin I and Semenogelin 11 fragments and soluble fibronectin fragments with liquefaction of the ejaculate and release of progressively motile spermatoza (Lilja, Laurell, C.B. (1984). Scand.

J. Clin. Lab. Invest. 44:447; McGee, Herr, J.C. (1987). Biol.

Reprod. 37:431). Furthermore, PSA may proteolytically degrade

IGFBP-

3 (insulin-like growth factor binding protein 3) allowing IGF to stimulate specifically the growth of PSA secreting cells (Cohen et al., (1992) J.

Clin. Endo. Meta. 75:1046-1053).

PSA complexed to alpha 1 antichymotrypsin is the predominant molecular form of serum PSA and may account for up to 95% of the detected serum PSA (Christensson, Bj6rk, Nilsson,

O.,

et al. (1993). J. Urol. 150:100-105; Lilja, Christensson, Dahl6n, U. (1991). Clin. Chem. 37:1618-1625; Stenman, Leinoven,

J.,

Alfthan, et al. (1991). Cancer Res. 51:222-226). The prostatic tissue (normal, benign hyperplastic, or malignant tissue) is implicated to predominantly release the mature, enzymatically active form of PSA, as this form is required for complex formation with alpha 1 antichymotrypsin (Mast, Enghild, Pizzo, et al. (1991).

Biochemistry 30:1723-1730; Perlmutter, Glover, Rivetna,

M.,

et al. (1990). Proc. Natl. Acad. Sci. USA 87:3753-3757). Therefore, in the microenvironment of prostatic PSA secreting cells, the PSA is believed to be processed and secreted in its mature enzymatically active form not complexed to any inhibitory molecule. PSA also forms stable complexes with alpha 2 macroglobulin, but as this results in encapsulation of PSA and complete loss of the PSA epitopes, the in vivo II -JC WO 97/14416 PCT/US96/ 1 6 4 9 0 -3significance of this complex formation is unclear. A free, noncomplexed form of PSA constitutes a minor fraction of the serum PSA (Christensson, Bjork, Nilsson, et al. (1993). J. Urol. 150:100- 105; Lilja, Christensson, Dahl6n, U. (1991). Clin. Chem.

37:1618-1625). The size of this form of serum PSA is similar to that of PSA in seminal fluid (Lilja, Christensson, Dahln, U. (1991).

Clin. Chem. 37:1618-1625) but it is yet unknown as to whether the free form of serum PSA may be a zymogen; an internally cleaved, inactive form of mature PSA; or PSA manifesting enzyme activity. However, it seems unlikely that the free form of serum PSA manifests enzyme activity, since there is considerable (100 to 1000 fold) molar excess of both unreacted alpha 1 antichymotrypsin and alpha 2 macroglobulin in serum as compared with the detected serum levels of the free 33 kDa form of PSA (Christensson, Bjbrk, Nilsson, et al. (1993).

J.

Urol. 150:100-105; Lilja, Christensson, Dahln, U. (1991). Clin.

Chem. 37:1618-1625).

Serum measurements of PSA are useful for monitoring the treatment of adenocarcinoma of the prostate (Duffy, M.S. (1989). Ann.

Clin. Biochem. 26:379-387; Brawer, M.K. and Lange, P.H. (1989).

Urol. Suppl. 5:11-16; Hara, M. and Kimura, H. (1989). J Lab. Clin.

Med. 113:541-548). Above normal serum concentrations of PSA have also been reported in benign prostatic hyperplasia and subsequent to surgical trauma of the prostate (Lilja, Christensson, Dahl6n,

U.

(1991). Clin. Chem. 37:1618-1625). Therefore, a cytotoxic compound that could be activated by the proteolytic activity of PSA should be prostate cell specific as well as specific for PSA secreting prostate metastases. Such a specific agent may be effective against BPH without causing the side-effects associated with other therapies.

Accordingly, it is the object of this invention to provide a novel pharmaceutical composition useful for the treatment of benign prostatic hyperplasia which comprises novel oligopeptides, which are selectively cleaved by enzymatically active PSA, in conjugation with a cytotoxic agent.

r C WO 97/14416 PCT/US96/16 4 9 0 -4- Another object of this invention is to provide a method of treating benign prostatic hyperplasia which comprises administration of the novel pharmaceutical composition.

SUMMARY OF THE INVENTION Novel pharmaceutical compositions useful for the treatment of adverse conditions of the prostate, in particular benign prostatic hyperplasia, which comprise novel oligopeptides, which are selectively cleaved by enzymatically active PSA, in conjugation with a pharmaceutical agent are described. Methods of treating such conditions of the prostate are also disclosed.

BRIEF DESCRIPTION OF THE FIGURES FIGURES 1 and 1A: Primary Amino Acid Sequence of Semenogelin

I:

The primary amino acid sequence of Semenogelin I is shown.

(SEQ.ID.NO.: 1) The PSA proteolytic cleavage sites are shown (numbered in order of the relative affinity of a site towards

PSA

hydrolysis) and the protein fragments are numbered sequentially starting at the amino terminus.

FIGURE 2: Cleavage Affinity of Synthetic Oligopeptides: A nested set of synthetic oligopeptides was prepared and the oligopeptides were digested with enzymatically active free PSA for various times. The results are shown in Table 2. All of the oligopeptides were tested as trifluoroacetate salts.

FIGURES 3, 3A and 3B: Cleavage Affinity of Synthetic Oligopeptides: Synthetic oligopeptides were prepared and the oligopeptides were digested with enzymatically active free PSA for four hours. The percentage of the oligopeptide that is cleaved in this period of time is listed. The results are shown in Table 4. Table 4a shows the amount of time (in minutes) required for 50% cleavage of the noted oligopeptides with enzymatically active free PSA. If no salt is indicated for an oligopeptide, the free base was tested.

WO 97/14416 PCT/US96/16490 FIGURE 4: Cytotoxicity Data ofNon-cleavable Oligopeptide- Doxorubicin Conjugates: The data of the figure shows comparative cytotoxicity of doxorubicin and a conjugate of doxorubicin covalently bound to an oligopeptide (Compound 12d) that does not contain the free PSA proteolytic cleavage site. The EC50 for doxorubicin is 0.3gM, while the acetylated oligopeptide modified doxorubicin has an EC50 that has been reduced by greater than 300 fold. This conjugate had no HPLC detectable contamination with unmodified doxorubicin. The oligopeptide alone had no detectable cell killing activity.

FIGURES 5 and 5A: Cleavage Affinity of Oligopeptides in Conjugation with Doxorubicin by Free PSA In Vitro: Qligopeptides-doxorubicin conjugates were prepared and the conjugates were digested with enzymatically active free PSA for four hours. The percentage conjugate that is enzymatically cleaved in the oligopeptide in this period of time is listed. The results are shown in Table 5. Table shows the amount of time (in minutes) required for 50% cleavage of the noted oligopeptide-cytotoxic agent conjugates with enzymatically active free PSA. If no salt is indicated for the conjugate, the free conjugate was tested.

FIGURE 6: Cleavage Affinity of Oligopeptides in Conjugation with Doxorubicin in Cell Conditioned Media: Oligopeptides-doxorubicin conjugates were reacted for four hours with cell culture media that had been conditioned by exposure to LNCaP cells (which are known to secrete free PSA) or DuPRO cell (which do not secrete free PSA). The percentage conjugate that is enzymatically cleaved in the oligopeptide in this period of time is listed. The results are shown in Table 6.

FIGURE 7: Cytotoxicity Data of Cleavable Oligopeptide-Doxorubicin Conjugates: WO 97/14416 PCT/US96/16 4 9 0 -6- The data in Table 7 shows cytotoxicity (as EC5 0 of conjugates of doxorubicin covalently bound to an oligopeptide that contain a free PSA proteolytic cleavage site against a cancer cell line that is known to secrete free PSA. Also shown for selected conjugates is the cytotoxicity of the conjugate against a cell line (DuPRO) which does not secrete free PSA.

If no salt is indicated for the conjugate, the free conjugate was tested.

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to pharmaceutical compositions that comprise conjugates that contain oligopeptides, which are specifically recognized by the free prostate specific antigen

(PSA)

and are capable of being proteolytically cleaved by the enzymatic activity of the free prostate specific antigen, and pharmaceutical agents covalently linked to such oligopeptides directly or through a linker unit, or pharmaceutically acceptable salts thereof. In particular, this invention is directed to such conjugates wherein the pharmaceutical agent is a cytotoxic agent. The present invention also relates to a novel method of treating adverse conditions of the prostate, in particular benign prostatic hyperplasia, which utilizes these compositions.

Such oligopeptides include oligomers that comprise an amino acid sequence selected from: a) AsnLysIleSerTyrGlnlSer (SEQ.ID.NO.: 13), b) LysIleSerTyrGlnlSer (SEQ.ID.NO.: 14), c) GlyGluAsnGlyValGlnLysAspValSerGlnXaaSerlleTyrlSerGlnThrGlu (SEQ.ID.NO.: d) GlyLysGlyIleSerSerGlnTyrlSerAsnThrGluGluArgLeu (SEQ.ID.NO.: 2), e) AsnLysIleSerTyrTyrlSer (SEQ.ID.NO.: 127), I WO 97/14416 PCT/US96/16 4 9 0 -7f) AsnLysAlaSerTyrGln1Ser (SEQ.ID.NO.: 128), g) SerTyrGIniSerSer (SEQ.ID.NO.: 129); h) LysTyrGIniSerSer (SEQ.JD.NO.: 140); i) hArgTyrGlSer5er (SEQ.JD.NO.: 14 1); j) hArgChaGln1Ser~er (SEQ.ID.No.: 185); and k) TyrGiniSerSer (SEQ.ID.NO.: 186); wherein hArg is homoarginine, Cha is cyclohexylalanine and Xaa is any natural amino acid.

In an embodiment of the instant invention, the oligopeptides include oligomers that comprise an amino acid sequence that is selected from: a) AsnLysIleSerTyrGlnlSerSer (SEQ-ID.NO.: 16), b) AsnLyslleSerTyrGln1SerAja (SEQ.ID.NO.: 130), c) AsnLyslleSerTyrGln1SerSerSer (SEQ.ID.No.: 17), d) AlaAsnLysIleSerTyrGlnlSerSerSe (SEQ.JD.No.: 18), e) LyslleSerTyrGIni SerSerSerThrGlu (SEQ.ID.No.: 19), f) Gl~us~yaGny~pa~rGnrSrl~r~r~~rl (SEQ.ID.NO.: 4), g) Gl~us~yaGny~pa~rGne~r~~r~r~~rl (SEQ.ID.NO.: WO 97/14416 PCT/US96/16 4 9

O

h) Al~ s~ y~l~e~y~y1S r SE .I .N-8-31) AlaAsnLysleSerTyrGyrlSer (SEQ.ID.NO.: 131), j) SerTyrGlnISerSerThr (SEQ.JD.NO.: 133), k) SerTyrGlnISerSerSer (SEQ.ID.NO.: 134), 1) LysTyrG1SerSer~er (SEQ.ID.NO.: 142), rn) h~gyGn1e~re (SEQ.ID.NO.: 143), and n) SerTyrGlnlSerSerLeu (SEQ.ID.NO.: 135); or the pharmaceutically acceptable salt thereof.

In a more preferred embodiment of the instant invention, the oligopeptides include oligomers that comprise an amino acid sequence that is selected from: a) AsnLyslleSerTyrGln iSerSerSerThr (SEQ.ID.NO.: b) AlaAsnLyslleSerTyrGln ISerAla (SEQ.ID.NO.: 136), c) AsnLysIeSerTyrGlnIS erSerSerThrGlu (SEQ.ID.No.:3) d) AlaAsnLyslleSerTyrGln ISerSerSerTlhrGlu (S EQ.ID.No.: 11), e) GIyGluAsnGIyVa1GnLysAspVaSrGlnrgSerlleTyrI SerGlnmhrGlu (SEQ.ID.NO.: 4), f) AlaAsnLyslleSerTyrTyr1SerSer (SEQ.ID.NO.: 137), WO 97/14416 PCT/US96/16490 -9g) AlaAsnLysIleSerTyrTyrlSerAla (SEQ.ID.NO.: 138), h) AlaAsnLysAlaSerTyrGlnlSerAla (SEQ.ID.NO.: 139), i) AlaSerTyrGlnlSerSerLeu (SEQ.ID.NO.: 94); or the pharmaceutically acceptable salt thereof.

In a further embodiment of the instant invention, the oligopeptides include oligomers that comprise an amino acid sequence The phrase "oligomers that comprise an amino acid sequence" as used hereinabove, and elsewhere in the Detailed Description of the Invention, describes oligomers of from about 6 to about 100 amino acids residues which include in their amino acid sequence the specific amino acid sequence decribed and which are therefore proteolytically cleaved within the amino acid sequence described by free PSA. Thus, for example, the following oligomer: GlnLeuAspAsnLysIleSerTyrGlnlSerSerSerThrHisGlnSerSer (SEQ.ID.NO.: 20) comprises the amino acid sequence: AsnLysIleSerTyrGlnlSerSerSerThr and would therefore come within the instant invention. It is understood that such oligomers do not include semenogelin I and semenogelin

II.

It is also understood that the instant invention includes oligomers wherein the N-terminus amino acid or the C-terminus amino acid, or both terminus amino acids are modified. Such modifications include, but are not limited to, acylation of the amine group at the Nterminus and formation of an amide to replace the carboxylic acid at the C-terminus. Addition of such moieties may be performed during solidphase synthesis of the oligomer; thus, attachment of the C-terminus ~a~ll WO 97/14416 PCTIUS96/1 6490 amino acid to a solid phase resin may be through an am-ine which results in an amide moiety upon acidic cleavage of the oligomer from the resin.

Thus the following compounds are considered itoligomers that comprise an amino acid sequence" as used hereinabove and are meant to be illustrative and are not limiting: Al~ny~ee~r~Ie~re~rl-md (SEQ.ID.NO.: 11) AcAas~s~~ryr~Ie~re~re (SEQ .ID. NO.: AcAas~s~~ryGnIe~re~rl-md (SEQ.ID.No.: 11) AcAas~s~~ryGnIe~re~re-md (SEQ.ID.No.: AcAas~s~~ryGnIe~ae~rl-md (SEQ.ID.NO.: 73) AcAas~s~~ryr Ie~ry~rl-amide (SEQ.ID 74) AcAas~sl~ryGn~re~rl-md (SEQ.ID.No.: AcAas~s~~ry~nIe~r~~rl-md (SEQ.ID 78) AcAas~s~~ryGnIe~ay~rl-md (S EQ.ID.NO. :79) Ac-AlaAsnLysIeS erTyrGlnISermbrGluamid (SEQ. ID.NO.: 81) AcAas~se~r~n~re~rl-md (SEQ. ID. NO.: 82) AcAas~sl~ryGnIe~ae~rl-md (SEQ.ID

.NO.:

84) AcAas~ul~ryGnIe~ae~rl-md (SEQ.ID.No.: Ac-AsnLysIeS erTyrGlnlSers er-amide (SEQ. ID. NO.: 16) Ac-LysIeSerTyrGlnISer~er-am~ide (SEQ.ID. NO.: 86) Ac-SerTyrGlnlS erSerThrGlu-amide (SEQ.ID 87) Ac-AlaSerTyrGlnlSerSerThrGlu-amide (SEQ. ID. NO.: 89) AcAas~sl~ry~rie~re~rl-md (SEQ .ID.NO.: 92) AcAas~s~~ry~rIe~ae~ri-md (SEQ.ID 93) Ac-AlaSerTyrGlnlSerSerLeu-amide (SEQ.ID 94) AcAas~ryGn~e~re~rl-md (SEQ.ID.NO.: AcAae~r~ie~re~rl-nid (SEQ.JD. NO.: 96) Ac-SerTyrGlnIS erSerSerThrGlu..amide (SEQ 97) AcAas~sl~e r~Ie~aerCys-amide (SEQ.ID.NO.: 98) Ac-hArg(Cha)Gln ISerNle-Acid (SEQ.ID.NO.: 147) Achrhy~n~re~eAi (SEQ.ID.NO.: 148) WO 97/14416 PCT/US96/1 6490 AchrhCaGn~re~eAi (SEQ.ID.NO.: 149) Ac-AlaAspLysAlaSerTyrGnIS erSer-Cha-.NHNH 2 (SEQ.JD.NO.: 150) Ac-hArgTyrGlnISerSerPr 0 Ac 1 d (SEQ.ID.NO.: 151) Ac-hArgTyrGlnISerSerJij 5 Acid (SEQ.JD.NO.: 152) Ac-hArgTyrGlnISerAsn-Acidj (SEQ.JD.NO.: 153) Ac-hArgoTyrG~nISer~ erSerNle-Acid (SEQ.ID. NO.: 154) Ac-(Amf)TyrGlnlSerSerSerNle-Acid (SEQ.ID.NO.: 155) H2C-~gyGn~e~re~uAi (SEQ. ID.NO.: 156) Ac-A~aAspLysAaLysTyrGIIIeSer~(Ch)NHNH2 (SEQ.ID. NO.: 157) Ac-(DPL)TyrGIniSerSerSerNle-Acid (SEQ.JD.NO.: 158) Ac-(imidazole)LysTyrG~nISerSerLeuAid (SEQ.JD.NO.: 159) AcAas~sl~~gTyGn~re~uAi (SEQ.ID.NO.: 160) Ac-(p-NH2Cha)TyrG~nIeSerSerN1Acd (SEQ.ID.NO.: 161) Ac (imidazolyl)LysTyrGlnISerSerSerNl -Acd (SEQ.ID. NO.: 162) Achr(h)~Ie~re~eAi (SEQ.ID.NO.: 163) Achr~r~Iere~r~gAi (SEQ.ID.NO.: 164) Ac-hArgTyrGlnISerSerSer(Me~eu) (SEQ.ID 188) erSer(Ethylester.Leu) (SEQ .ID. NO.: 156) Ac-AaAspLysAa(imidoeLys)TGISSNleAi

(SEQ.ID.NO.:

165) Achro(-oo-y) ie~rerNie-Acid (SEQ.ID.No.: 166) Ac-hArg(Me2PO3jyr)G1flj5er erSerNle-Acid (SEQ.ID.No.: 167) Ac-hArgTyrGlnISerSerAsp..Acjd (SEQ.ID.No.: 168) g( TrGn~r~re~eAi (SEQ.ID.No.: 169) Ac-AlaAspLysAaLysTyrG~niSerSeNlAid (SEQ.JD 170) Achra(h)~ie~re~tyetrLu (SEQ.ID.No.: 171) Ac-(imidazolyl)Lys (Cha)GlnlSerSerSerNle-Acid (SEQ.ID.No.: 172) Achr(h)~I~re~rAi (SEQ.JD 173) Achr(h)lI~re~eAi (SEQ. JD.NO.: 174) Achr,(h)~n~rr~eAi (SEQ.ID.No.: 175) and Achr~-loo-y) Ie~rerNie-Acid (SEQ.ID.No.: 176), or the pharmaceutically acceptable salt thereof.

WO 97/14416 PCT/US96/16490 -12- A person of ordinary skill in the peptide chemistry art would readily appreciate that certain amino acids in a biologically active oligopeptide may be replaced by other homologous, isosteric and/or isoelectronic amino acids wherein the biological activity of the original oligopeptide has been conserved in the modified oligopeptide Certain unnatural and modified natural amino acids may also be utilized to replace the corresponding natural amino acid in the oligopeptides of the instant invention. Thus, for example, tyrosine may be replaced by 3iodotyrosine, 2 -methyltyrosine, 3 -fluorotyrosine, 3 -methyltyrosine and the like. Further for example, lysine may be replaced with imidazolyl)lysine and the like. The following list of amino acid replacements is meant to be illustrative and is not limiting: Original Amino Acid Ala Arg Asn Asp Glu Gin Gly Ile Leu Lys Met Omithine Phe Ser Thr Trp Tyr Val Replacement Amino Acid(s) Gly Lys, Omithine Gln Glu Asp Asn Ala Val, Leu, Met, Nle lie, Val, Met, Nle Arg, Omithine Leu, Ile, Nle, Val Lys, Arg Tyr, Trp Thr Ser Phe, Tyr Phe, Trp Leu, Ile, Met, Nle WO 97/14416 PCT/IUS96/1 6 4 9 0 -13 Thus, for example, the following oligopeptides may be synthesized by techniques well known to persons of ordinary skill in the art and would be expected to be proteolytically cleaved by free PSA: AsnArgIleSerTyrGnI5er (SEQ.JD.NO.: 21) AsnLysValSerTyrG 1 1 J5er (SEQ.ID.NO.: 22) AsnLysMetS erTyrGlnl SerSer (SEQ.JD.No.: 23) AsnLysLeuSerTyr~ln ISerSer (SEQ. ID. NO.: 24) As~s~~ryrlIe~re (SEQ. ID. NO.: As~s~~rhe~Ie~re (SEQ.JD. NO.: 26) As~s~~rr~n~re~rh (SEQ.ID.NO.: 27) As~s~~ryrsIe rerThr (SEQ. ID. NO.: 28) As~s~~ryrlIh rerThr (SE Q.ID.NO.: 29) As~s~~r~rlIe (SEQ. ID.NO.: I~nyIee~y~n~re (SEQ.ID.NO.: 31) As~gl~ryr~Ie~re (S EQ.JD. NO.: 32) As~g~~rhGn~re~rh (SEQ. ID. NO.: 33) As~gl~rr~n~re~rh (SEQ.ID. NO.: AsnArgfleSerTyrGflnI~hr erS erThr (SEQ .ID.NO.: 36) As~s~~ry~n~re~rh (SEQ .ID. NO.: 37) AsnLysLeuSerTyrGl 1 1 IThrSerSerThr (SEQ. ID. NO.: 38) G~~se~ryGn~re~rh (SEQ. ID. NO.: 39) AsnArgLeuSerTyrGlnj ThrSerSerThr (SEQ. ID.NO.: As~sa~rhGn~re~rh (SEQ.ID.NO.: 41) As~ga~rr~lle~re'b (SEQ.ID.NO.: 42) GlnLysValSerTyrGl 11 ISerS erSer'fhr (SEQ.ID 43) GlnLysIleSerTyrGlnI~hr erSerThr (SEQ. ID 34) As~s~~ryr Ie~rerThr (SEQ.ID.NO.: 44); or the pharmaceutically acceptable salt thereof.

Similarly, the following oligopeptides may be synthesized by techniques well known to persons of ordinary skill in the art and would be expected to be proteolytically cleaved by free

PSA:

WO 97/14416 PCT/US 96/1 6490 14 GlYGIuGInGIY VaIGIIILysAspVaiSer ~erIleTyrISerGlfl~p1,Glu (SEQ.ID-NO.: GIYGuAsnGyLeuGInLysAspVlSGIn r11 leTyrIS crGInThrGlu (SEQ.ID.NO.: 47), GlyG~uAsnGIyVaAsLysAsVlSG nerS cr11eTyriS erGlnThrGlu (SEQ.JD.NO.: 48), GIyG1uAsnGyVaGnArgAVal rGnJ c~r1 IeTyrIS erGlnThrGlu (SEQ.ID-NO.: 49), GlYGuAsnGIYVa1GulnLysAsVlSGInyS cr1 eTvriS erGlnThrGlu (SEQ-ID.NO.: GIYGuAsnGyVaIGInLysAsLSG ~r11 leTyrI S rGlnThrGlu (SEQ.ID.NO.: 51), GIYGIuAsnGly ValGInLysAspVIlSerG r11 e~e~eGn~rl (SEQ.ID.NO.: 52), GlYGuAsnGyVaIGInLysAMSerG ~r1 IleTyrIThrG~nThrGlu (SEQ.ID.NO.: 53), cr11 eTyrlThrGlnThrGlu (SEQ.ID.NO.: 54), GIyGluAsnG~yVa1GIIILysAspVIS erGInSerS cr1 IeTyri SerGInS erGlu (SEQ.ID.NO.: Gl~usn~ c~ny~paleGn r11 rleTyrIS erAsnThrGlu (SEQ.ID.NO.: 56), GIyLysAlaIleScrSerGlnTyrI ScrAsnThrGluGI uArgcLcu

(SEQ.ID.NO.:

57), ArgcLeu (SEQ

.JD.NO.:

59), GlyLysGlyllcThrSerGInTyrISers r~ul~rre (SEQ.ID.

NO.:

GlyLysG1ylleSrT11rGnTyrISAThGlulrge (SEQ.JD.

NO.:

GlyLysGlylleS crSerAsnTyrISrAsnTrGuGlu~roLe

(SEQ.ID.NO.:

WO 97/14416 PCTIJS96/1 6 4 9 0 AlaLysGlylleS erSerGInTyrl SerAsnThrGluGlu~rgLeu GlyLysGlylleSerSerGnPheSerAsnThrGluGlu~rgLeu 64), GlyLysGlylleSerSerGnTyrIThrAsnhGluGlu~rgLeu GlyLysG~yleSerSerGlnTyrISerAsnSerGluGluArgoLeu 58), and GlyLysGlylleS erSerG~nTyrISerAsnThrAspGlu~rcgLe 46); and the like.

(SEQ.ID

NO.:

(SEQ.ID.NO.:

(SEQ.ID

NO.:

(SEQ-ID

NO.:

(SEQ.ID

NO.:

The inclusion of the symbol within an amino acid sequence indicates the point within that sequence where the oligopeptide is Proteolytically cleaved by free PSA.

The compounds of the present invention may have asymmetric centers and occur as racemates, racemic mixtures, and as individual diastereomers, with all possible isomers, including optical isomers, being included in the present invention. Unless otherwise specified, named amino acids are understood to have the natural

"L"

stereoconfiguration The following abbreviations are utilized in the specification and figures to denote the indicated amino acids and moieties: hR or hArg: hY or hTyr: Cha: Amf:

DPL:

(imidazolyl)K: Me2PO3-Y: O-Me-Y:

TIC:

MeL

DAP:

homoarginine homotyrosine cyclohexylalanine 4-mnmtyphnllnn 6 -dimethylpyrinidinyl)lysine 2 -iniidazolyl)lysine O-dimethylphosphotyrosine O-methyltyrosine tetrahYdro-3isoquinoline carboxylic acid 2-eo3aio5mtyhxn 1, 3 -diaminopropane WO 97/14416 PCT/US96/164 9 0 -16- TFA: trifluoroacetic acid AA: acetic acid The method of treatment of the instant invention utilizes pharmaceutical compositions whose pharmaceutical activity is specific for cells that secrete enzymatically active PSA. Such compositions comprise the oligopeptides described herein above covalently bonded directly, or through a linker unit, to a pharmaceutical agent. Such a combination of an oligopeptide and pharmaceutical agent may be termed a conjugate. The pharmaceutical agent component of the conjugate may be selected from known compounds useful for treating conditions of the prostate, whose site of biological activity or the desired target of the biological activity is within the prostate or in close proximity to the prostate. Such pharmaceutical agents include, but are not limited to cytotoxic agents.

In a preferred embodiment, the method of treatment of the instant invention utilizes cytotoxic compositions whose cytotoxicity is specific for cells that secrete enzymatically active PSA. Such compositions comprise the oligopeptides, described herein above, covalently bonded directly, or through a linker unit, to a cytotoxic agent.

Ideally, the cytotoxic activity of the cytotoxic agent is greatly reduced or absent when the oligopeptide containing the PSA proteolytic cleavage site is bonded directly, or through a chemical linker, to the cytotoxic agent and is intact. Also ideally, the cytotoxic activity of the cytotoxic agent increases significantly or returns to the activity of the unmodified cytotoxic agent upon proteolytic cleavage of the attached oligopeptide at the cleavage site. While it is not necessary for practicing this aspect of the invention, a preferred embodiment of this aspect of the invention is a conjugate wherein the oligopeptide, and the linker unit if present, are detached from the cytotoxic agent by the proteolytic activity of the free PSA and any other native proteolytic enzymes present in the tissue proximity, thereby releasing unmodified cytotoxic agent into the i. WO 97/14416 PCT/US96/16490 -17physiological environment at the place of proteolytic cleavage.

Pharmaceutically acceptable salts of the conjugates are also included It is understood that the oligopeptide of the instant invention that is conjugated to the cytotoxic agent, whether through a direct covalent bond or through a linker unit, does not need to be the oligopeptide that has the greatest recognition by free PSA and is most readily proteolytically cleaved by free PSA. Thus, the oligopeptide that is selected for incorporation in such an anti-BPH composition will be chosen both for its selective, proteolytic cleavage by free PSA and for the cytotoxic activity of the cytotoxic agent-proteolytic residue conjugate (or, in what is felt to be an ideal situation, the unmodified cytotoxic agent) which results from such a cleavage.

Because the conjugates utilized in the instant invention can be used for modifying a given biological response, cytotoxic agent is not to be construed as limited to classical chemical therapeutic agents. For example, the cytotoxic agent may be a protein or polypeptide possessing a desired biological activity. Such proteins may include, for example, a toxin such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin; a protein such as tumor necrosis factor, a-interferon, P-interferon, nerve growth factor, platelet derived growth factor, tissue plasminogen activator; or, biological response modifiers such as, for example, lymphokines, interleukin- interleukin-2 interleukin-6 granulocyte macrophage colony stimulating factor

("GM-

2 CSF"), granulocyte colony stimulating factor or other growth factors.

The preferred cytotoxic agents include, in general, alkylating agents, antiproliferative agents, tubulin binding agents and the like.

Preferred classes of cytotoxic agents include, for example, the anthracycline family of drugs, the vinca drugs, the mitomycins, the bleomycins, the cytotoxic nucleosides, the pteridine family of drugs, diynenes, the taxanes and the podophyllotoxins. Particularly useful members of those classes include, for example, doxorubicin carminomycin, daunorubicin, aminopterin, methotrexate, methopterin, dichloro-methotrexate, mitomycin C, porfiromycin, 5-fluorouracil, 6- WO 97/14416 PCT/US96/1 6 4 9 0 -18mercaptopurine, cytosine arabinoside, podophyllotoxin, or podophyllotoxin derivatives such as etoposide or etoposide phosphate, melphalan, vinblastine, vincristine, leurosidine, vindesine, leurosine, taxol and the like. Other useful cytotoxic agents include estramustine cisplatin and cyclophosphamide. One skilled in the art may make chemical modifications to the desired cytotoxic agent in order to make reactions of that compound more convenient for purposes of preparing conjugates of the invention.

A highly preferred group of cytotoxic agents for the present invention include drugs of the following formulae: THE METHOTREXAT GROUP OF FORMULAl

H

2 N N N

R

8

COR

9 N N

CONHCHCH

2

CH

2

CO

2

H

R

2

R

8 (1) in which

R

12 is amino or hydroxy;

R

7 is hydrogen or methyl;

R

8 is hydrogen, fluoro, chloro, bromo or iodo;

R

9 is hydroxy or a moiety which completes a salt of the carboxylic acid; WO 97/14416 PCT/US96/164 9 0 19 THE MITOMYCIN GROUP OF FRMULA.(2:

H

3

C

COH

2

OCONH

2 0OCH 3 -N R 10 in which RIO is hydrogen or methyl; THE BLEOMYCIN GROUp OFT T- lFORMULA (3) H .AOJJ'JIr 2 0 HO CHH 0N

H

2 N 0 0YN NH N

N

CH

3 N 0

I

H N-

OH

3 H HO OH 3 HO0

'N

HO

OH 0

OH

H1 0 0

OH

CONH

2 (3) in which Rll is hydroxy, amino, CI-C3 alkylamino, di(C1-C 3 allcyl)amino, C4-C6 polymethylene amino, WO 97/14416 PCTJUS96/1 6 4 9 0 20

NH

-NHCH

2

CH

2

CH

2

S-CH

3 or -NHCH 2

CH

2

CH

2

CHNHCN

CH

3 MELPHALAN OF FORMULLA (41: HO -CH-0H 2 NC 2

CH

2

CI)

2

NH

2 (4) 6 -M ERCAkPTOPURINE OF FORMULA SH

H

N

'N N~ A CY(TOSINE ARABINO)SIDE OFT FOMUL A (6:

NH

2

NO

HOH

2 C 0 1 HO

OH

(6) WO 97/14416 PCTIUS961 6490 -21- THE PODOP~i-YL-LOTOXINS OF FORMULA(7): R14 0 0~-

CH

3

O'

0

OH

iR which R 13 is hydrogen or methyl;

R

14 is methyl or thienyl; or a phosphate salt thereof; WO 97/14416 PCT/US96/16490 -22- THE VINCA ALKALOID GROUP OF DRUGS OF FORMULA

R'

6 N

"'R

17 NN ,,R18 N CO 2

CH

3

H

1

N

""'CH

2

CH

3 N OR 19

CO

2

CH

3 in which

R

15 is H, CH3 or CHO; when R1 7 and R 1 8 are taken singly;

R

18 is H, and one of R 16 and R 17 is ethyl and the other is H or OH; when R1 7 and R 18 are taken together with the carbons to which they are attached, they form an oxirane ring in which case R 16 is ethyl;

R

19 is hydrogen, (C1-C3 alkyl)-CO, or chlorosubstituted (C1-C3 alkyl)-CO; DIFLUORONUCLEOSIDES OF FORMULA R21 O CH 2

OH

F

in which

R

2 1 is a base of one of the formulae: WO 97/14416 PCT/US96/16490 23 o 0 HNHN N 0 H 2 N N N

NH

2 NH 2 0 N KN N in which

R

2 2 is hydrogen, methyl, bromo, fluoro, chioro or iodo;

R

2 3 is -OH or -NH-2;

R

2 4 is hydrogen, bromo, chioro or iodo; or, THE ANTHRACYCLINES ANTIBIOTICS OF FORMULA(1) wherein RI is -CH3, -CH2OH, -CH2OCO(CH2)3CH 3 or 2 H5) 2 WO 97/14416 PCT/US96/16490 24-

R

3 is -OCH3, -OH or -H;

R

4 is -NH2, -NHCOCF3, 4 -morpholinyl, 3 -cyano-4morpholinyl, 1-piperidinyl, 4-methoxy-l-piperidinyl, benzylamine, dibenzylamine, cyanomethylamine, or 1cyano-2-methoxyethyl amine; is -OH -OTHP or and

R

6 is -OH or -H provided that

R

6 is not -OH when R 5 is -OH or -OTHP.

ESTRAMUSTINE (11) CH3 OH

(CICH

2

CH

2 2

NCOO

(11) CYCLOPHOSPHAMIDE (12) 0 .N(CH 2

CH

2

CI)

2 f 0

NH

12 The most highly preferred drugs are the anthracycline antiobiotic agents of Formula described previously. One skilled in the art understands that this structural formula includes compounds which are drugs, or are derivatives of drugs, which have acquired in the art different generic or trivial names. Table 1, which follows, represents a number of anthracycline drugs and their generic or trivial names and which are especially preferred for use in the present invention.

Table 1 Compound daunorubicina doxorubicinb detorubicin carminomycin idarubicin epirubicin esorubicin

THP

AD-32 Ra CR3 CH20H CH2OCOCH(OC 2

H

5 2 CM3 CH3 CH20H CH20H CH20H CH2OCO(CH- 2 3 CR3 Rb OCH3 OCH3 OCH3

OH

H

OC113 OCH3 OCH3

RC

NH2 N112 NH2 NH2 NH2 NH2 OHf OHf Off OHf OHf

H

OTHP

H

OH

a~~daunomycin"OCH isaNHtrntvenmOfrdFnruii badamycinll is an alternative name for dauorubicin ~ft WO 97/14416 PCT/US96/16490 -26- Of the compounds shown in Table 1, the most highly preferred cytotoxic agents are doxorubicin, vinblastine and desacetylvinblastine. Doxorubicin (also referred to herein as "DOX") is that anthracycline of Formula (10) in which RI is -CH20H, R3 is -OCH3, R4 is -NH2, R5 is -OH, and R6 is -H.

The oligopeptides, peptide subunits and peptide derivatives (also termed "peptides") incorporated in the conjugates utilized in the method of treatment of the present invention can be synthesized from their constituent amino acids by conventional peptide synthesis techniques, preferably by solid-phase technology. The peptides are then purified by reverse-phase high performance liquid chromatography

(HPLC).

Standard methods of peptide synthesis are disclosed, for example, in the following works: Schroeder et al., "The Peptides", Vol. I, Academic Press 1965; Bodansky et al., "Peptide Synthesis", Interscience Publishers, 1966; McOmie "Protective Groups in Organic Chemistry", Plenum Press, 1973; Barany et al., "The Peptides: Analysis, Synthesis, Biology" 2, Chapter 1, Academic Press, 1980, and Stewart et al., "Solid Phase Peptide Synthesis", Second Edition, Pierce Chemical Company, 1984. The teachings of these works are hereby incorporated by reference.

The pharmaceutically acceptable salts of the compounds incorporated in the conjugates utilized in the method of treatment of this invention include the conventional non-toxic salts of the compounds of this invention as formed, from non-toxic inorganic or organic acids. For example, such conventional nontoxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like: and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenyl-acetic, glutamic, benzoic, salicylic, sulfanilic, 2 -acetoxy-benzoic, fumaric, WO 97/14416 PCT/US96/16490 -27toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic and the like.

The conjugates utilized in the method of treatment of the instant invention which comprise the oligopeptide containing the PSA cleavage site and a cytotoxic agent may similarly be synthesized by techniques well known in the medicinal chemistry art. For example, a free amine moiety on the cytotoxic agent may be covalently attached to the oligopeptide at the carboxyl terminus such that an amide bond is formed. Similarly, an amide bond may be formed by covalently coupling an amine moiety of the oligopeptide and a carboxyl moiety of the cytotoxic agent. For these purposes a reagent such as 2-(1Hbenzotriazol-1 -yl)-1, 3 3 -tetramethyluronium hexafluorophosphate (known as HBTU) and 1-hyroxybenzotriazole hydrate (known as HOBT), dicyclohexyl- carbodiimide (DCC), N-ethyl-N-(3dimethylaminopropyl)- carbodiimide (EDC), diphenylphosphorylazide (DPPA), benzotriazol-1-yl-oxy-tris-(dimethylamino)phosphonium hexafluorophosphate (BOP) and the like, used in combination or singularly, may be utilized.

Furthermore, the instant conjugate may be formed by a nonpeptidyl bond between the PSA cleavage site and a cytotoxic agent. For example, the cytotoxic agent may be covalently attached to the carboxyl terminus of the oligopeptide via a hydroxyl moiety on the cytotoxic agent, thereby forming an ester linkage. For this purpose a reagent such as a combination of HBTU and HOBT, a combination of BOP and imidazole, a combination of DCC and DMAP, and the like may be utilized. The carboxylic acid may also be activated by forming the nitrophenyl ester or the like and reacted in the presence of DBU (1,8diazabicyclo[5,4,0]undec-7-ene.

The instant conjugate may also be formed by attachment of the oligopeptide to the cytotoxic agent via a linker unit. Such linker units include, for example, a biscarbonyl alkyl diradical whereby an amine moiety on the cytotoxic agent is connected with the linker unit to form an amide bond and the amino terminus of the oligopeptide is connected with the other end of the linker unit also forming an amide bond. Conversely, WO 97/14416 PCT/US96/16490 -28a diaminoalkyl diradical linker unit, whereby a carbonyl moiety on the cyctotoxic agent is covalently attacted to one of the amines of the linker unit while the other amine of the linker unit is covalently attached to the C terminus of the oligopeptide, may also be uselful. Other such linker units which are stable to the physiological environment when not in the presence of free PSA, but are cleavable upon the cleavage of the PSA proteolytic cleavage site, are also envisioned. Furthermore, linker units may be utilized that, upon cleavage of the PSA proteolytic cleavage site, remain attached to the cytotoxic agent but do not significantly decrease the cytotoxic activity of such a post-cleavage cytotoxic agent derivative when compared with an unmodified cytotoxic agent.

One skilled in the art understands that in the synthesis of conjugates utilized in the method of treatment of the invention, one may need to protect or block various reactive functionalities on the starting cbmpounds and intermediates while a desired reaction is carried out on other portions of the molecule. After the desired reactions are complete, or at any desired time, normally such protecting groups will be removed by, for example, hydrolytic or hydrogenolytic means. Such protection and deprotection steps are conventional in organic chemistry. One skilled in the art is referred to Protective Groups in Organic Chemistry, McOmie, ed., Plenum Press, NY, NY (1973); and, Protective Groups in Organic Synthesis, Greene, ed., John Wiley Sons, NY, NY (1981) for the teaching of protective groups which may be useful in the preparation of compounds of the present invention.

By way of example only, useful amino-protecting groups may include, for example, CI-C10 alkanoyl groups such as formyl, acetyl, dichloroacetyl, propionyl, hexanoyl, 3 3 -diethylhexanoyl, ychlorobutryl, and the like; C1-C10 alkoxycarbonyl and C5-C15 aryloxycarbonyl groups such as tert-butoxycarbonyl, benzyloxycarbonyl, allyloxycarbonyl, 4 -nitrobenzyloxycarbonyl, fluorenylmethyloxycarbonyl and cinnamoyloxycarbonyl; halo-(C1-C10)-alkoxycarbonyl such as 2,2, 2 -trichloroethoxycarbonyl; and Cl-C15 arylalkyl and alkenyl group such as benzyl, phenethyl, allyl, trityl, and the like. Other commonly WO 97/14416 PCT/US96/16490 -29used amino-protecting groups are those in the form of enamines prepared with P-keto-esters such as methyl or ethyl acetoacetate.

Useful carboxy-protecting groups may include, for example, alkyl groups such as methyl, tert-butyl, decyl; halo-C1-C10 alkyl such as 2 ,2,2-trichloroethyl, and 2-iodoethyl; C5-C15 arylalkyl such as benzyl, 4-methoxybenzyl, 4 -nitrobenzyl, triphenylmethyl, diphenylmethyl; Cl-C 10 alkanoyloxymethyl such as acetoxymethyl, propionoxymethyl and the like; and groups such as phenacyl, 4halophenacyl, allyl, dimethylallyl, tri-(C1-C3 alkyl)silyl, such as trimethylsilyl, P-p-toluenesulfonylethyl, P-p-nitrophenyl-thioethyl, 2,4,6trimethylbenzyl, P-methylthioethyl, phthalimidomethyl 2,4-dinitrophenylsulphenyl, 2 -nitrobenzhydryl and related groups.

Similarly, useful hydroxy protecting groups may include, for example, the formyl group, the chloroacetyl group, the benzyl group, the b-enzhydryl group, the trityl group, the 4 -nitrobenzyl group, the trimethylsilyl group, the phenacyl group, the tert-butyl group, the methoxymethyl group, the tetrahydropyranyl group, and the like.

With respect to the preferred embodiment of the instant method of treatment in which an oligopeptide is combined with the anthracycline antibiotic doxorubicin, the following Reaction Schemes illustrate the synthsis of the conjugates of the instant invention.

WO 97/14416 PCTIUS96,1 6490 30 REACTION SCHEME I

CH

2 0H OH 0

CH

2 0H dox

CH

3 0

CH

3

OH

C-terminus

OH

)-Eolpeptide

I

C-terminus

NH-

2 WO 97/14416 PCT/U5S96/16490 -31 REACTION SCHEME 11

CH

3 0

CH

3

NH-

2 11 OH NH-protect /C-terminus

OH

HN -Lol~opeptide

CH

2 0H

'OH

CH

3 0

OH

WO 97/14416 PCT/US96/16490 32 REACTION SCHEME III

CH

2 0H

CH

3 0 0y(C H 2 2 00 2

H

OH

HN

OH

00" 0 jo g p

OCH

2 0H bH N-terminus

OH

3

NH-

2

OH

WO 97/14416 PCTIUS96/16490 33 REACTION SCHEME IV

CH

3 0

OH

11

H

2 NHN -ojigpeptide

I

C-terminus C-terminus N =oigpeptide

CH

3 0

CH

3

OH

16

NH-

2 WO 97/14416 PCT/US96/16490 34 REACTION SCHEME V Br 2 CC1 4

CH

2 Br

CH

3 0

CH

3

NH-

2

OH

HS

0

H

C-terminus

OH

CH

3 0

CH

3 N ojigopeptide C-terminus

NH

2 WO 97/14416 PCT/US96/16490 Reaction Scheme VI illustrates preparation of conjugates utilized in the instant method of treatment wherein the oligopeptides are combined with the vinca alkaloid cytotoxic agent vinblastine.

Attachment of the N-terminus of the oligopeptide to vinblastine is illustrated Kandukuri et al. J. Med. Chem. 28:1079-1088 (1985)).

Reaction Scheme VII illustrates preparation of conjugates utilized in the instant method of treatment wherein the oligopeptides are combined with the vinca alkaloid cytotoxic agent vinblastine wherein the attachment of vinblastine is at the C-terminus of the oligopeptide. The use of the 1, 3 -diaminopropane linker is illustrative only; other spacer units between the carbonyl of vinblastine and the C-terminus of the oligopeptide are also envisioned. Furthermore, Scheme VII illustrates a synthesis of conjugates wherein the C-4-position hydroxy moiety is reacetylated following the addition of the linker unit. Applicants have discovered that the desacetyl vinblastine conjugate is also efficacious and may be prepared by eliminating the steps shown in Reaction Scheme

VII

of protecting the primary amine of the linker and reacting the intermediate with acetic anhydride, followed by deprotection of the amine. Conjugation of the oligopeptide at other positions and functional groups of vinblastine may be readily accomplished by one of ordinary skill in the art and is also expected to provide compounds useful in the treatment of benign prostatic hyperplasia.

It is also understood that conjugates may be prepared wherein the N-terminus of the oligopeptide utilized in the instant method of treatment is combined with one cytotoxic agent, such as vinblastine, while the C-terminus is simultaneously attached to another cytotoxic agent, which is the same or different cytotoxic agent, such as doxorubicin. Reaction Scheme VIII illustrates the synthesis of such a polycytotoxic agent conjugate. Such a polycytotoxic conjugate may offer advantages over a conjugate containing only one cytotoxic agent.

WO 97/14416 PCT/rJS96/I 6490 36 REACTION SCHEME

VI

OH

-N

%%E

"H NAH, reflux, MeOH vinblastine 00 2

CH

3

HONO

CH

3 O0 N

CH

3 0'

CONHNH

2 CON. 3 WO 97/14416 PCT/US96/1 6490 -37- REACTION SCHEME VI (Continued)

OH

C-terminus 1. oligopeptide

R

2. AC 2 O, pyridine

CH:

3 0

CON

3

CH

3

O.

C-terminus wherein R is -NH 2 -0-alkyl and the like WO 97/14416 PCTIUS96/1649 0 38 REACTION SCHEME

VII

6H 3 1. H 2 N CH 2

CH

2

OH

2

NH

2 2. BOO-Cl "'0H 2 0H 3 3 1. AC 2 O, pyridine 2. aq. HOI H -BOO 0H 2 0H 3

NOCOCH

3 WO 97/14416 PCT/US96/16490 39 REACTION SCHEME VII (Continued) oligopeptidle

I

C-terminus

CH

2

CH

3

NOCOCH

3 N- oligopeptidle

R'

C-terminus wherein R' is acetyl, alkyl, hydrogen or the like WO 97/14416 PCT/US96,I 6490 REACTION SCHEME VIII Et 6H 3 oligopeptide 2. Ac 2 O, pyridine or 0-terminus 3~ 1. oligopeptide 00H 3

"CH

2

CH

3 2. LiOH I *OH 3. AC 2 O, pyridine

OH

3 doxorubicin

DCH

3 ptide

I

C-terminus I

CH

3 oligope; WO 97/14416 PCTIUJS96,I 6490 -41- REACTION SCHEME VIII (Continued)

I

N-terminus

CH

2 0H 0 OH 0 I m I WO 97/14416 PCT/US96/1649 0 -42- The oligopeptide-cytotoxic agent conjugate utilized in the method of treatment of the instant invention wherein the cytotoxic agent is the preferred cytotoxic agent doxorubicin may be described by the general formula I below: O OH 0

SCHOH

SOH

0 OH

O

CH

3

O

NH

OH XL oligopeptide

R

wherein: oligopeptide is an oligopeptide which is specifically recognized by the free prostate specific antigen (PSA) and is capable of being proteolytically cleaved by the enzymatic activity of the free prostate specific antigen; XL is absent or is an amino acid selected from: a) phenylalanine, b) leucine, c) valine, d) isoleucine, e) 2 -naphthyl)alanine, f) cyclohexylalanine, g) diphenylalanine, h) norvaline, i) norleucine, and WO 97/14416 PCT/US96/1649 0 -43j) 1,2,3 4 -tetrahydroiso quinoline-3 -carboxylic acid; R is hydrogen or 1; and R I is C I-C6-alcyl or aryl, or the pharmaceutically acceptable salt thereof.

In a preferred embodiment of the instant method of treatment of BPH: oligopeptide is an oligomer that comprises an amino acid sequence selected from: a) AsnLyslleSerTyrGln1Ser (SEQ.JD.No.: 13), b) LyslleSerTyrGiniSer (SEQ.ID.No.: 14), c) GlyGluAsnG~yVaIG~nLysAspVaSerG1naaS erIeTyrIS erGlnThrGlu (SEQ.ID.NO.: 15) d) GlyLysGlyeSerSerG~nTyrISerAsnThrGluGlu~rgLeu (SEQ.ID.NO.: 2), e) AsnLyslleSerTyrTyrlSer (SEQ.ID.No.: 127), f) AsnLysAlaSerTyrGlnlSer (SEQ.ID.NO.: 128), 1g) SerTyrGlnlSerSer (SEQ.JD.NO.: 129), h) LysTyrGIniSerSer (SEQ.ID.NO.: 14); i) hArgTyrGlnlSerSer (SEQ.ID.NO.: 141); WO 97/14416 PCT/US9616490 -44j) hArgChaGlnlSerSer k) TyrGlnlSerSer (SEQ.ID.NO.: 185); and SEQ.ID.NO.: 186); wherein Xaa is any natural amino acid; XL is absent or is an amino acid selected from: a) leucine, b) isoleucine, c) norleucine, and d) valine; and R is acetyl, pivaloyl or benzoyl, or the pharmaceutically acceptable salt thereof.

The following compounds are specific examples of the oligopeptide-cytotoxic agent conjugate utilized in the method of treatment of the instant invention: WO 97/14416 PCT/US96/16490

CH

2 0H

CH

3 0 wherein X is: AsnLyslleSerTyrGlnSer AsnLyslleSerTyrGlnSerSer- AsnLyslleSerTyrGlnSerSerSer AsnLyslleSerTyrGlnSerSerSerThr AsnLyslleSerTyrGlnSerSerSerThrGlu AiaAsnLyslleSerTyrGlnSerSerSerThrGlu- N-terminus (SEQ.ID.NO.: 13), (SEQ.ID.NO.: 16), (SEQ.ID.NO.:17), (SEQ.ID.NO.:1O), (SEQ.ID.NO.: 3), (SEQ.ID.NO.: 11) WO 97/14416 PCT/US96/16490 46 Ac AaAsnLysIleSerTyrG InSerSerSerThr- Ac AlaAsnLysleSerTyrGlnSerSerSerTh rLeu Ac -AlaAsnLysAlaSerTyrGInSerAlaSerThrLeu Ac, AlaAsnLysAaSerTyrGinSerAaSerLeu-.

Ac AlaAsnLysAfaSerTyrGlnSerSerSerLeu- Ac AlaAsnLysAaSerTyrGnSerSerLeu- Ac -SerTyrGInSerSerSerLeu

(SE

Ac hArgTyrGIneSerSereu

(SE

Ac LysTyrGnSerSerSerLeu-

(SE

(Co Ac-LysTyrGnSerSefie (SEt (SEQ.ID.NO.: 117), (SEQ.ID.NO.: (SEQ.JD.NO.: 118), (SEQ.ID.NO.: 119), (SEQ.ID.NO.: 120), (SEQ.ID.NO.: 121).

Q.ID.NO.: 144), Q.ID.NO.: 145).

Q.ID.NO.: 124), or mpound 4) Q.ID.NO.: 146).

N-terminus or the pharmaceutically acceptable salt thereof.

Further examples of conjugates of an oligopeptide and doxorubicin wherein the N-terminus of the oligopeptide is acylated and the C-terminus of the oligopeptide is attached to the doxorubicin at the 3'amine are as follows: Ac-hArgTyrGln-.SerS erPro-dox(3') (SEQ.ID 151) Ac-hArgTyrGln-SerPro-dox(3') (SEQ.JD 177) Ac-hArgTyrGln-SerSerSerNle-dox(3I) (SEQ.JD 154) -47 Ac-AmfTyrGln-S erS erSerNle-dox(3 t) (SEQ.ID.NO.: 155) H2NCO-hArgTyrG~n..serserSerLeu.dox(3f) (SEQ.ID.No.: 156) Ac-LysTyrGln-SerSerNle..dox (SEQ.ID.NO.: 146) Ac-LysTyrGln-SerLysNle-.dox(3') (SEQ. ID 178) Ac(cis-p-NH2Cha)TyrGnSerSerNledo(3') (SEQ.ID.No.: 161) Ac-AlaAspLysAla(h1Jrg)TyrGln..SerS erLeu-dox (SEQ.ID. NO.: 160) Ac-hArgaTyrGln-SerAsn-dox(3') (SEQ.ID.NO.: 153) Ac-hArgTyrGln-S erS erHis-dox(3') (SEQ.ID.NO.: 152) Ac-(imidazolyl)LysTyrGlnSerSerLeu-dox(3') (SEQ.ID.No.: 159) Ac-(imidazolyl)LysTyrGlnSerS erSerNle-dox(3') (SEQ.ID.N~o: 162) Ac-hArg,(Cha)Gln..SerSerSerNle-dox(3I) (SEQ.ID.NO.: 163) Ac-hArg,(Me2PO3Tyr)oln..SerS erSerNle-dox(3') (SEQ.ID.NO.: 167) Ac-flArgyrGIn-SerSerSerh~rgdox(3) (SEQ.JD.NO. 14) Ac-h~a(j-loao-yr)JlnSerSerSerNle-dox(3') (SEQ.ID.NO. 166t) Ac-hArg(O-Me-Tyr)Gln-SerSerSerNle-.dox(3?) (SEQJID.NO.: 169) Achr~-H-h)l-Sre~rl-o(' (SEQ.ID.NO.: 179) Ac-hArg(Cha)Gln-SerSerNe-dox(3') (SEQ.ID.NO.: 174) Ac-hArg(Cha)Gln-SerProNle..dox(3') (SEQ. JD.NO.: 175) Ac(imidazolyl)Lys(Cha)GlnSerS erSerNle-dox(3') (SEQ.ID.No.: 172) Ac-hArg(7HOrTIC)Gln..SerS erS erNle-dox(3') (SEQ.ID.NO.: 180) Ac.-hArg(3-Fluoro)TyrG~nS erSerS erNle-dox(3') (SEQ.ID.NO.: 176) Ac-(ornithine)TyrGln..SerSerS erNle-dox(3') (SEQ.ID.NO.: 181) Ac-LysAlaAlaSerSerSerLeu-.dox(3') (SEQ.ID.NO.: 182) Ac-hArgh(Cha)Gln-S erS erNle-dox(3') (SEQ.ID.NO.: 149) Ac-AlaArgLysAlaSerTyrG~n-SerLeu-do(3 2 (SEQ.ID.NO.: 193) and Ac-(Orn)TyrGlw.SerSerSerLeu.dox(3r) (SEQ.ID.NO.: 194) or the pharmaceutically acceptable salt thereof.

The oligopeptide-cytotoxic agent conjugate utilized in the method of treatment of the instant invention wherein the cytotoxic agent is the preferred cytotoxic agent vinbiastine or desacetylvinbiastine may be described by the general formula I below: WO 97/14416 PCTIUS96/16490 -48

CH

2

CH

3 'rOR1 9

CH

3 0' XL oligopeptide R wherein: oligopeptide is an oligopeptide which is specifically recognized by the free prostate specific antigen (PSA) and is capable of being proteolytically cleaved by the enzymatic activity of the free prostate specific antigen; XL is absent or is an amino acid selected from: a) phenylalanine, b) leucine, c) valine, d) isoleucine, e) (2-naphthyl)alanine, f) cyclohexylalanine, g) diphenylalanine, h) norvaline, and i) norleucine, and j) 1 2 3 4 -tetrahydroisoquinoline-3-carboxylic acid; or XL is NH -(CH2)n -NH WO 97/14416 PCT/US96/16490 -49- R is hydrogen or -(C=O)R1; R1 is C1-C6-alkyl or aryl;

R

19 is hydrogen or acetyl; and n is 1,2, 3, 4 or or the pharmaceutically acceptable salt thereof.

The following compounds are specific examples of the oligopeptide-desacetylvinblastine conjugate utilized in the method of treatment of the instant invention:

'OH

u NH N-terminus Ac- LysTyrGlnSerSerSerNe

NH

Compound 14 (SEQ.ID.NO.: 183), WO 97/14416 PCT/US961649 0

OH

CH

2

CH

3 C-terminus LeuAsnLysAlaSerTyrGInSerSerSerLeu

NH

2 Compound 5 (SEQ.ID.NO.: 184), or the pharmaceutically acceptable salt thereof.

The following compounds is a specific example of the polycytotoxic agent conjugates utilized in the method of treatment of the instant invention: WO 97/14416 PCT/US96/16 4 90 -51-

OH

,LEt CH3 CHaO SC=O C-terminus LeuAsnLysAlaSerTyrGInSerSerLeu (SEQ.D.NO.: 184), OH (SEQ.ID.NO.: 184),

CH

3 Compound

CH

2

OH

0 or the pharmaceutically acceptable salt thereof.

It is well known in the art, and understood in the instant invention, that peptidyl therapeutic agents such as the oligopeptidecytotoxic agent conjugates preferably have the terminal amino moiety of any oligopeptide substituent protected with a suitable protecting group, such as acetyl, benzoyl, pivaloyl and the like. Such protection of the terminal amino group reduces or eliminates the enzymatic degradation of such peptidyl therapeutic agents by the action of exogenous amino WO 97/14416 PCT/US96/16490 -52peptidases which are present in the blood plasma of warm blooded animals.

The oligopeptide-cytotoxic agent conjugates utilized in the method of treatment of the instant invention are administered to the patient in the form of a pharmaceutical composition which comprises a conjugate of Formula and a pharmaceutically acceptable carrier, excipient or diluent therefor. As used, "pharmaceutically acceptable" refers to those agents which are useful in the treatment or diagnosis of a warm-blooded animal including, for example, a human, equine, procine, bovine, murine, canine, feline, or other mammal, as well as an avian or other warm-blooded animal. The preferred mode of administration is parenterally, particularly by the intravenous, intramuscular, subcutaneous, intraperitoneal, or intralymphatic route. Such formulations can be prepared using carriers, diluents or excipients familiar to one skilled in the art. In this regard, See, e Remington's Pharmaceutical Sciences, 16th ed., 1980, Mack Publishing Company, edited by Osol et al. Such compositions may include proteins, such as serum proteins, for example, human serum albumin, buffers or buffering substances such as phosphates, other salts, or electrolytes, and the like. Suitable diluents may include, for example, sterile water, isotonic saline, dilute aqueous dextrose, a polyhydric alcohol or mixtures of such alcohols, for example, glycerin, propylene glycol, polyethylene glycol and the like. The compositions may contain preservatives such as phenethyl alcohol, methyl and propyl parabens, thimerosal, and the like. If desired, the composition can include about 0.05 to about .20 percent by weight of an antioxidant such as sodium metabisulfite or sodium bisulfite.

For intravenous administration, the composition preferably will be prepared so that the amount administered to the patient will be from about .01 to about 1 g of the conjugate. Preferably, the amount administered will be in the range of about .2 g to about 1 g of the conjugate. The conjugates of the invention are effective over a wide dosage range depending on factors such as the disease state to be treated or the biological effect to be modified, the manner in which the conjugate is administered, the age, weight and condition of the patient as well as WO 97/14416 PCT/US96/16490 -53other factors to be determined by the treating physician. Thus, the amount administered to any given patient must be determined on an individual basis.

One skilled in the art will appreciate that although specific reagents and reaction conditions are outlined in the following examples, modification can be made which are meant to be encompassed by the spirit and scope of the invention. The following preparations and examples, therefore, are provided to further illustrate the invention, and are not limiting.

EXAMPLES

EXAMPLE 1 Identification of the Semenogelin PSA Mediated Cleavage Site: Liquefaction of the seminal gel parallels proteolytic fragmentation of semenogelin I [Lilja, Laurell, (1984) Scand. J. Clin. Lab. Inves.

44, 447-452]. It is believed that the proteolytic fragmentation of semenogelin is mainly due to the proteolytic activity of prostate-specific antigen [Lilja, (1985) J. Clin. Invest. 76, 1899-1903]. Utilizing the published sequence of semenogelin I [Lilja, Abrahamsson,

P.A.,

Lundwall, (1989) J. of Biol. Chem. 264, 1894-1900] (Figure 1) we designed polymerase chain reaction primers to clone the semenogelin cDNA from a commercially available prostatic cDNA library (Clonetech, Palo Alto, The purified semenogelin cDNA was placed into the bacterial expression vector pTAC [Linemeyer, Kelly,

L.J.,

Minke, Gimenez-Gallego, DeSalvo, J. and Thomas,

K.A.,

(1987) Bio/Technology 5, 960-965]. The semenogelin cDNA was designed so that a tubulin epitope was placed at the carboxyl end of semenogelin protein.. The bacterially expressed semenogelin protein was purified on an anti-tubulin antibody column. The purified semenogelin

I

protein was mixed with commercially prepared prostate-specific antigen (PSA) (York Biologicals International, Stony Brook, NY) in an 100 to 1 molar ratio (semenogelin I/PSA) in 12 mM Tris pH 8.0, 25 mM NaCI, WO 97/14416 PCT/US96/1 6 4 9 0 -54mM CaC12, and incubated for various times. The digest was fractionated by polyacrylamide gel electrophoresis and transferred by electrophoresis to ProBlott filter paper (Applied Biosystems, Inc., Foster City, CA.) in CAPS buffer [Matsudaira, (1987) J. Biol. Chem. 252, 10035-10038]. The ProBlott filter paper was stained with coomassie blue to identify the novel PSA generated semenogelin I protein fragments.

The novel fragments were cut out of the filter with a scalpel and submitted for sequence determination. After the proteolytic fragments were identified by variable time digestion, a 10 minute digestion reaction was performed. The affinity of PSA for the 5 potential cleavage sites in semenogelin I was determined to be as follows: site 349/350 site 375/376 site 289/290 site 315/316 site 159/160. The relative affinities were derived from the comassie blue staining intensity of each PSA generated peptide fragment. These intensities had approximate ratios of 3:1:0.6:0.3.

EXAMPLE 2 Preparation of Oligopeptides which Comprise the PSA Mediated Cleavage Site: Oligopeptides were prepared by solid-phase synthesis, using a double coupling protocol for the introduction of amino acids on the Applied Biosystems model 430A automated peptide synthesizer. Deprotection and removal of the oligopeptide from the resin support were achieved by treatment with liquid hydrofluoric acid. The oligopeptides were purified by preparative high pressure liquid chromatography on reverse phase C18 silica columns using an aqueous 0.1% trifluoroacetic acid/acetonitrile gradient. Identity and homogeneity of the oligopeptides were confirmed by amino acid composition analysis, high pressure liquid chromatography, and fast atom bombardment mass spectral analysis.

The oligopeptides that were prepared by this method are shown in Figure 2.

I I M WO 97/14416 PCT/US96/164 9 0 EXAMPLE 3 Assessment of the Recognition of Oligopeptides by Free PSA The oligopeptides prepared as described in Example 2 were individually dissolved in PSA digestion buffer (12 mM tris(hydroxymethyl)aminomethane pH8.0, 25 mM NaC1, 0.5 mM CaC12) and the solution added to PSA at a molar ration of 100 to 1. Alternatively, the PSA digestion buffer utilized is 50 mM tris(hydroxymethyl)-aminomethane pH7.4, 140 mM NaC1. The reaction is quenched after various reaction times by the addition of trifluoroacetic acid (TFA) to a final 1% (volume/volume). Alternatively the reaction is quenched with ZnCl2. The quenched reaction was analyzed by HPLC on a reversedphase C18 column using an aqueous 0.1 %TFA/acetonitrile gradient. The results of the assessment are shown in Figure 2. Other oligopeptides prepared as described in Example 2 were tested in the same assay wherein the reaction was quenched at 4 hours. Those results of the assessment are shown in Figure 3. The removal of an asparagine residue from the amino terminus of the oligopeptide results in a significant loss of PSA mediated peptide hydrolysis, while the presence of a glutamic acid residue at the carboxyl end of the peptide appears not to be essential to recognition by PSA.

EXAMPLE 4 Preparation of Non-cleavable Oligopeptide-Doxorubicin Conjugates: The derivatives of doxorubicin shown in Table 3 were prepared using the following general reaction: To a mixture of doxorubicin (Sigma) and the corresponding peptide (prepared by solid phase synthesis or commercially available (Sigma)) in DMSO was added HBTU and HOBT along with diisopropylethylamine and the reaction mixture was stirred overnight. The crude reaction mixture was purified directly by preparative HPLC on a reversed-phase C-18 column using a 0.1% trifluoroacetic acid (TFA) in acetonitrile/0.1% TFA in water gradient.

WO 97/14416 PCT/US96/1649 0 -56- Table 3

CH

2 0H

'OH

CH

3 0 Compound 12a 12b 12c 12d

R

H-Ala- N-Ac-Ala- N-Ac-Ala-Ala-Ala- N-Ac-Ala-Gly-Pro-Thr-Gly-Ala-Ser- Ala- MS (parent ion) 615 657 799.5 1199 (SEQ.ID.NO.: 12) EXAMPLE In vitro Assay of Cytotoxicity of Peptidyl Derivatives of Doxorubicin: The cytotoxicities of the non-cleaveable oligopeptide-doxorubicin conjugates, prepared as described in Example 4, against a line of cells which is known to be killed by unmodified doxorubicin were assessed with an Alamar Blue assay. Specifically, cell cultures of LNCaP prostate tumor cells, which are a human metastatic prostate adenocarcinoma isolated from a needle biopsy of a lymph node (LNCaP.FGC: American Type Culture Collection, ATCC CRL 1740), or DuPRO cells in 96 well plates were diluted with medium containing various concentrations of a given conjugate (final plate well volume of 200tl). The cells were WO 97/14416 PCT/US96/16490 -57incubated for 3 days at 37 0 C and then 20 l of Alamar Blue was added to the assay well. The cells were further incubated and the assay plates were read on a EL-310 ELISA reader at the dual wavelengths of 570 and 600 nm at 4 and 7 hours after addition of Alamar Blue. Relative percentage viability at the various concentration of conjugate tested was then calculated versus control (no conjugate) cultures. Cytotoxicities of unmodified doxorubicin and unmodified oligopeptide were also assessed.

Figure 3 shows the cytotoxicity data for a representative compound (Compound 12d).

EXAMPLE 6 Assessment of Enzymatically Active PSA from LNCaP Cells Enzymatic activity was demonstrated by incubating LNCaP serum free media (concentrated approximately 200 fold) with recombinant Sememogelin I protein. Approximately 0.5 pg of immunologically reactive PSA in concentrated conditioned media [determined by HYBRIDTECH (Tandem E) elisa] was mixed with approximately 3 gg of recombinant Semenogelin I and incubated for 4 hours at 37 0 C. At the end of the incubation, the digest mixture was analyzed by Western blot procedures. The results show that purified PSA from semen and PSA from LNCaP conditioned media generate identical proteolytic maps of the recombinant Semenogelin I protein. Thus, LNCaP cells produce enzymatically active PSA. LNCaP are tumorigenic in nude mice and produce detectable levels of circulating

PSA.

EXAMPLE 7 Preparation of Cleavable Oligopeptide-Doxorubicin Conjugates: The derivatives of doxorubicin wherein an oligopeptide which is proteolytically cleaved by free PSA is covalently attached to the amine of the sugar moiety of the doxorubicin were prepared using the following general reaction: To a mixture of doxorubicin (Sigma) and the corresponding peptide (prepared by solid phase synthesis as described in WO 97/14416 PCT/US96/16490 -58- Example 2) in DMSO was added HBTU and HOBT along with diisopropylethylamine and the reaction mixture stirred overnight. The crude reaction mixture was purified directly by preparative HPLC on a reversed-phase C-18 column using a 0.1% trifluoroacetic acid (TFA) in acetonitrile/0.1% TFA in water gradient. When reactive amine moieties were present on the peptide, such a functionality was typically protected as the fluorenylmethyloxycarbonyl adduct, which was removed by treatment with a secondary amine, such as piperidine and the like, subsequent to conjugation with doxirubicin. The instant conjugates have a structure of the general formula 0 OH 0

CH

2 0H CHO3 r

NH

OH

(O=C)peptide(NH) Ac and may be represented by the phrase "Ac-peptide-DOX Conjugates which were prepared by the above general method or by the synthetic route described in Example 8, but utilizing the appropriate starting amino acid residues which are readily available commercially or by synthetic techniques well known in the art, are listed in Tables 5, and 7 in Figures 5, 5A and 7.

EXAMPLE 8 Ac-Lvs-Tyr-Gln-Ser-Ser-Ser-Leu-Dox*Acetate WO 97/14416 PCT/US96/16490 -59- Step A: Ac-Lys(Fmoc)-Gln-Ser(Bzl)-Ser(Bzl)-Ser(Bzl)-Leu-PAM Resin Starting with 0.5 mmol (0.67g) Boc-Leu-PAM resin, the protected peptide was synthesized on a 430A ABI peptide synthesizer.

The protocol used a 4 fold excess (2 mmol) of each of the following protected amino acids: Boc-Ser(OBzl), Boc-Gln, Boc-Tyr(BrZ), Boc- Lys(Fmoc). Coupling was achieved using DCC and HOBT activation in methyl-2-pyrrolidinone. Acetic acid was used for the introduction of the N terminal acetyl group. Removal of the Boc group was performed using 50% TFA in methylene chloride and the TFA salt neutralized with diisopropylethylamine. At the completion of the synthesis, the peptide resin was dried to yield 1.3g of Step B: Ac-Lvs(Fmoc)-Tvr-Gln-Ser-Ser-Ser-Leu-OH (2) The protected peptide resin 1.3 g, was treated with HF ml) for 2 hrs at 0°C in the presence of anisole (2 ml). After evaporation of the HF, the residue was washed with ether, filtered and extracted with DMF. The DMF filtrate (75 ml) was concentrated to dryness and triturated with H 2 0. The insoluble product was filtered and dried (0.46g).

Step C: Ac-Lys(Fmoc)-Tvr-Gln-Se-r-Ser-Leu-Dox (3) The above prepared intermediate 0.46g, (0.43 mmol) was dissolved in DMF (15 ml) and doxorubicin hydrochloride, 125 mg (0.215 mmol), added followed by 60 tl of triethylamine (0.430 mmol).

The stirred solution was cooled and 92 pl of diphenylphosphoryl azide (0.43 mmol) added. After 5 minutes, an additional 92 pl of DPPA was added and the pH adjusted to -7.5 (pH paper) with TEA. After 1 hour, an additional 92 p of DPPA was added, pH adjusted to and the reaction stirred at 0"-5°C overnight. After 18 hours, the reaction (found to be complete by analytical HPLC) was concentrated to an oil Step D: Ac-Lys-Gln-Tyr-Ser-Ser-Ser-Leu-Dox WO 97/14416 PCT/US9616490 The above product was dissolved in DMF (20 ml), cooled (0OC) and 10 ml of piperidine added. The solution was concentrated to dryness and purified by preparative HPLC. Buffer A acetic acid-H 2 0; B 15% acetic acid-methanol. The crude product was dissolved in 300 ml of 10% B/90% A buffer, filtered and purified on a C-18 reverse phase HPLC radial compression column (Waters, Delta- Pak 15gm, 300A). A step gradient of 10% B to 60% B was used at a flow rate of 75 ml/min (uv 260 nm). Homogeneous product fractions were pooled, concentrated and freeze-dried from H 2 0 to yield 125 mg of purified product EXAMPLE 9 Deacetylvinblastinyl-Leu-Asn-Lys-Ala-Ser-Try-Gln-Ser-Ser-Ser-Leu- NH2*Acetate (SEQ.ID.NO. 184) Step A: NH2 Leu-Asn-Lys(Fmoc)-Ala-Ser-Tyr-Gln-Ser-Ser-Ser- Leu-Amide (6) Starting with 0.5 mmol of p-methylbenzhydrylamine resin (MBHA), the protected peptide, NH2-Leu-Asn-Lys(Fmoc)-Ala- Ser(OBzl)-Tyr(BrZ)-Gln-Ser(OBzl)-Ser(OBzl)-Ser(OBzl)-Leu-MBHA, intermediate was synthesized on a 430A ABI peptide synthesizer. The protocol used a 4 fold excess (2 mmol) of each of the following protected amino acids: Boc-Leu, Boc-Asn, Boc-Lys (Fmoc), Boc-Ala, Boc- Ser(OBzl), Boc-Tyr(BrZ), Boc-Gln. Coupling was achieved using DCC and HOBT activation in N-methyl-2-pyrrolidinone

(NMP).

Removal of the Boc group was performed using 50% TFA in methylene chloride and the TFA salt neutralized with diisopropylethylamine. The dried protected peptide resin (1.80g) was treated with HF (20 ml) for 2 hrs at 0° C in the presence of anisole (2 ml).

After evaporation, the residue was extracted with DMF. The DMF filtrate (75 ml) was concentrated to dryness, dissolved in a 1:1 mixture of and freeze-dried to give 750 mg of crude product. A WO 97/14416 PCT/US96/1649 0 -61portion (200 mg) was purified by preparative HPLC on a C-18 reverse phase support (Waters, g-Bondapak). Buffer A 15% acetic B 15% acetic acid-methanol. For the purification, the crude product was suspended in 400 ml of 10% B/90% A buffer, filtered and the filtrate loaded onto the column. A step gradient of 10% B to 55% B was used at a flow rate of 75 ml/min. Homogeneous product fractions were pooled, concentrated and freeze-dried from H20 to yield Step B: Deacetvivinblastin Monohydrazide (7) 1g of vinblastine sulfate was converted to the amine form by extraction in methylene chloride and saturated sodium bicarbonate. The methylene chloride layer was washed with H20, dried over anhydrous MgSO4 and concentrated to dryness. The vinblastine was then dissolved in anhydrous ethanol (20 ml) and anhydrous hydrazine added (20 ml).

The solution was heated (60° C) under an N2 atmosphere for 17 hrs. The reaction was concentrated to an oil, dissolved in methylene chloride, extracted with H20 and dried over MgSO4. After evaporation compound was isolated. [Ref: K.S.P. Bhushana Rao et al. J. Med.

Chem. (1985), 28:1079.] Step C: Deacetvlvinblastine Acid Azide Deacetylvinblastine monohydrazide (48 mg, 0.0624 mmol) was dissolved in DMF (3 ml), cooled C) and acidified to (pH paper) with HCl/dioxane. Isoamylnitrite (10 pl) was added followed by an additional 10 gl after 10 min. HPLC analysis indicated complete conversion of the hydrazide to azide after 5 min. The azide was maintained in solution at -15 C until ready for use.

Step D: Deacetylvinblastinyl-Leu-Asn-Lys-Ala-Ser-Try-Gn-Ser- Ser-Ser-Leu-NH_.Acetate The oligopeptide product from Step A, 32 mg (0.0225 mmol), was dissolved in DMF (1 ml) and cooled To this solution was added a 1.5 ml DMF solution (0.031 mmol) of desacetylvinblastine acid azide The pH was adjusted to 7.5 (pH WO 97/14416 PCT/US96/164 9 0 62paper) with triethylamine and the reaction stirred at C (2 hr), and 0" C for 18 hr. To the reaction was added H20 (2 ml) and the solution evaporated to dryness. The intermediate was dissolved in DMF (4 ml), cooled C) and 2 ml of piperidine added. The solution was concentrated to dryness and purified by preparative HPLC as described in Step A. The homogeneous fractions were pooled, concentrated and freeze-dried from H20 to yield EXAMPLE Deacetylvinblastinyl-Leu-Asn-Lys-Ala-Ser-Try-Gln-Ser-Ser-Ser-Leu-- Dox *Acetate Step A: Deacetylvinblastinyl-Leu-Asn-Lys(Fmoc)-Ala Ser-Try-Gln-Ser-Ser-Ser-Leu-Dox Acetate (9) The oligopeptide product prepared as described in Example 9, Step A, (166 mg, 0.125 mmol), was dissolved in DMSO (3 ml) and cooled to -15° C. To this solution was added a DMF solution (0.125 mmol) of desacetylvinblastine acid azide prepared as described in Example 9, Step C. The pH was adjusted to 7.5 (pH paper) with triethylamine and the reaction stirred at -15° C for 90 mins.

After stirring 18 hours at 0-50 C, the reaction was concentrated to dryness and the crude residue was dissolved in DMF ml) and filtered. Doxorubicin hydrochloride, 62 mg (0.106 mmol), was added to the filtrate followed by 30 pl of triethylamine. The stirred solution was cooled and 27 pl of diphenylphosphoryl azide (DPPA, 0.134 mmol) added. After 5 minutes, an additional 27 p1 of DPPA was added and the pH adjusted to -7.5 (pH paper) with TEA. After 1 hour, an additional 27 tl of DPPA was added, pH adjusted to and the reaction stirred at 0"-5C overnight. After 18 hours, the reaction (found to be complete by analytical HPLC) was concentrated to an oil Step B: Deacetylvinblastinyl-Leu-Asn-Lys-Ala-Ser-Try- Gln-Ser-Ser-Ser-Leu--Dox Acetate WO 97/14416 PCT/US96/1 6 4 9 0 -63- The above intermediate product was dissolved in DMF ml), cooled and 10 ml of piperidine added. The solution was concentrated to dryness and purified by preparative HPLC. Buffer A acetic acid-H20; B 15% acetic acid-methanol. The crude product was dissolved in 300 ml of 10% B/90% A buffer, filtered and purified on a C-18 reverse phase HPLC radial compression column (Waters,

A-

Bondapak). A step gradient of 10% B to 60% B was used at a flow rate of 75 ml/mm (uv 260 nm). Semi-pure product was further purified on C-18 (Waters, Prep Pak) using Buffer A 0.13M pH triethylammonium phosphate and Buffer B acetonitrile. A ste gradient of 10% B to 40% B was used at a flow rate of 75 ml/min. (uv 214 nm). Pure product fractions were pooled, diluted with H20 and desalted by applying the product onto the same column and eluting the product as the actetate salt with 90% acetonitrile/10% H20 acetic acid). The product fractions were concentrated and freeze dried from to yield the purified product EXAMPLE 11 Ac-Lys-Tyr-Gln-Ser-Ser-Ser-Nle-NH-(CH2)3

NH-

deacetylvinblastine amide (14) Step A Deacetylvinblastine-3-aminopropvl amide (11) To a cooled C) a DMF solution (3 ml, 0.0624 mmol) of deacetylvinblastine acid azide (synthesis described in Example 9, Step C) was added 120 gl of 1, 3 -diaminopropane in DMF (2 ml). The reaction was stirred at 10" C for 1 hr, filtered and concentrated to dryness to yield (11).

StepB Deacetylvinblastine-3-aminopropylamidenorleucine amide (12 To a DMF solution (1 ml) of Boc-Nle (22 mg, 0.095 mmol) was added 318 1g of a 1M solution of HOBT (in NMP) followed by 280 p1 of a 1M solution of DCC (in NMP). After 30 min., intermediate (11) WO 97/14416 PCT/US96/16490 -64- (0.0624 mmol) was added in a 3.5 ml DMF. The pH of the reaction was adjusted 7.5 with diisopropylethylamine. After stirring for 18 hrs the reaction was concentrated to an oil and the Boc protecting group removed by treating the oil with a 1:1 solution of TFA: CH2C12 (20 ml). After min. the reaction was concentrated to dryness. Purification was achieved by preparative HPLC on a C-18 reverse phase support (Waters, Delta Pak). Buffer A 0.1% TFA-H20; B= 0.1% TFA-CH3CN. The crude product was loaded in 100% A buffer (100 mi) and a step gradient of 100% A to 30% A was used at a flow rate of 75 ml/min. Homogeneous product fractions were pooled and freeze-dried to yield (12).

Step C: Ac-Ls(Fmoc)-Tr-Gln-Ser-Ser-Ser-Nle-OH (13) The above intermediate was prepared as described in Example 9, Step Afor the preparation of Ac-Lys(Fmoc)-Tyr-Gln-Ser-Ser- Ser-Leu-OH.

Step D: Ac-Lys-Tyr-Gln-Ser-Ser-Ser-Nle-NH-(CH 2 3

NH-

deacetvlvinblastine amide (14) The oligopeptide product (70 mg, 0.065 mmol) in DMF (1 ml) was combined with (41 mg, 0.05 mmol) of (12) in DMF (4 ml). The solution was cooled C) and 17 .1 of diphenylphosphoryl azide (0.08 mmol) added. After 5 min. an additional 17 ll of DPPA was added and the pH adjusted to 7.5 (pH paper) with triethylamine. After 2 hr. additional 35 mg, was added in DMF (0.5 ml) and 17 41 of DPPA. The pH was maintained at 7.5 with TEA and after 3 hr. an additional 35 mg of (13) was added in DMF (0.5 ml). The reaction was stirred at 0-5" C.

After 18 hrs, the reaction was concentrated to dryness, redissolved in DMF (9 ml), cooled C) and 3 ml of piperidine added. The solution was concentrated to dryness and purified by preparative HPLC. Buffer A 0.1% TFA-H20; B= 0.1% TFA-CH3 CN. The crude product was dissolved in 30% acetic acid H20 (100 ml) and purified on a C-18 reverse phase HPLC radial compression column (Waters, Delta Pak). A step gradient of 100% A to 70% A was used at a flow rate of 75 ml/min.

Semi-pure product fractions were pooled and freeze-dried. Purification WO 97/14416 PCT/US96/16490 to homogeneity was achieved by repurification on a C-4 support (Waters, Delta Pak) as described above. Product fractions were pooled and freeze dried to yield pure (14).

EXAMPLE 12 Assessment of the Recognition of Oligopeptide-Doxorubicin Conjugates by Free PSA The conjugates prepared as described in Examples 7-9 were individually dissolved in PSA digestion buffer (12 mM tris(hydroxymethyl)aminomethane pH8.0, 25 mM NaC1, 0.5 mM CaC12) and the solution added to PSA at a molar ration of 100 to 1. Alternatively, the PSA digestion buffer utilized is 50 mM tris(hydroxymethyl)-aminomethane pH7.4, 140 mM NaC1. The reaction is quenched after various reaction times by the addition of trifluoroacetic acid (TFA) to a final 1% (volume/volume). Alternatively the reaction is quenched with ZnC12. The quenched reaction was analyzed by HPLC on a reversedphase C18 column using an aqueous 0.1 %TFA/acetonitrile gradient. The results of the assessment are shown in Tables 5 and 5a of Figure EXAMPLE 13 Assessment of the Cleavage of Oligopeptide-Doxorubicin Conjugates in Cell Conditioned Media Cell conditioned serum-free o-MEM media (phenol red minus) was collected 3 days after the addition of the media to either LNCaP or DuPRO (prepared as described in J. Urology, 146:915-919 (1991)) cell lines. The media was concentrated 20 fold using an Amicon® CentriprepTM concentrator with a 10,000 molecular weight cutoff. The LNCaP conditioned media contained free PSA protein at, on average, approximately 100 ng/mL concentration as determined by the Tandem®- E PSA immunodetection kit (Hybritech®). There was no detectable free PSA in the DuPRO cell conditioned media.

WO 97/14416 PCTIUS96/16490 -66- 100 gL portions of concentrated conditioned media was mixed with gg of a oligopeptide-doxorubicin conjugate prepared as described in Example 7 and the mixture was incubated at 37 0 C for 0, 4 and 24 hour time points. The reactions were stopped by the addition of ZnC12 (to a 0.01M final concentration) and analyzed by HPLC on a reversed-phase C18 column using an aqueous 0.1 %TFA/acetonitrile gradient to determine the percentage of peptide-cytotoxic agent conjugate that had been digested. The results of the assessment are shown in Table 6 of Figure 6.

EXAMPLE 14 In vitro Assay of Cytotoxicity of Peptidyl Derivatives of Doxorubicin: The cytotoxicities of the cleaveable oligopeptide-doxorubicin conjugates, prepared as described in Example 7, against a line of cells which is known to be killed by unmodified doxorubicin was assessed with an Alamar Blue assay as described in Example 5. Specifically, cell cultures of LNCaP prostate tumor cells or DuPRO cells in 96 well plates was diluted with medium containing various concentrations of a given conjugate (final plate well volume of 200g1). The cells were incubated for 3 days at 37°C, 2 0l of Alamar Blue is added to the assay well. The cells were further incubated and the assay plates were read on a EL-310 ELISA reader at the dual wavelengths of 570 and 600 nm at 4 and 7 hours after addition of Alamar Blue. Relative percentage viability at the various concentration of conjugate tested was then calculated versus control (no conjugate) cultures. Cytotoxicities of the conjugates were also compared to the cytotoxicity of unmodified doxorubicin and unmodified oligopeptide assessed in the same assay. Results of this assay are shown in Table 7 of Figure 7.

WO 97/14416 PCT/US96/16490 67- EXAMPLE In vivo Efficacy ofPeptidyl -Cytotoxic Agent Conjugates LNCaP.FGC or DuPRO-1 cells are trypsinized, resuspended in the growth medium and centifuged for 6 mins. at 2 00xg. The cells are resuspended in serum-free a-MEM and counted. The appropriate volume of this solution containing the desired number of cells is then transferred to a conical centrifuge tube, centrifuged as before and resuspended in the appropriate volume of a cold 1:1 mixture of a-MEM- Matrigel. The suspension is kept on ice until the animals are inoculated.

Male nude mice (10-12 weeks old) are restrained without anesthesia and are inoculated with 0.5 mL of cell suspension on the left flank by subcutaneous injection using a 22G needle. Mice are either given approximately 5x10 5 DuPRO cells or 1.5x10 7 LNCaP.FGC cells.

Following inoculation with the tumor cells the mice are treated under one of two protocols: Protocol A: One day after cell inoculation the animals are dosed with a 0.1-0.5 mL volume of test conjugate, doxorubicin or vehicle control (sterile water).

Dosages of the conjugate and doxorubicin are initially the maximum nonlethal amount, but may be subsequently titrated lower. Identical doses are administered at 24 hour intervals for 5 days. After 10 days, blood samples are removed from the mice and the serum level of PSA is determined. Similar serum PSA levels are determined at 5-10 day intervals. At the end of 5.5 weeks the mice are sacrificed and weights of any tumors present are measured and serum PSA again determined.The animals' weights are determined at the beginning and end of the assay.

Protocol B: Ten days after cell inoculation,blood samples are removed from the animals and serum levels of PSA are determined. Animals are then grouped according to their PSA serum levels. At 14-15 days after cell WO 97/14416 PCT/US96/16490 68inoculation, the animals are dosed with a 0.1-0.5 mL volume of test conjugate, doxorubicin or vehicle control (sterile water). Dosages of the conjugate and doxorubicin are initially the maximum non-lethal amount, but may be subsequently titrated lower. Identical doses are administered at 24 hour intervals for 5 days. Serum PSA levels are determined at 5-10 day intervals. At the end of 5.5 weeks the mice are sacrificed, weights of any tumors present are measured and serum PSA again determined. The animals' weights are determined at the beginning and end of the assay.

WO 97/14416 PCT/US96/16490 -69- SEQUENCE LISTING GENERAL INFORMATION: APPLICANT: DeFeo-Jones, Deborah Garsky, Victor M.

Jones, Raymond E.

Oliff, Allen I.

Scolnick, Edward M.

(ii) TITLE OF INVENTION: CONJUGATES USEFUL IN THE TREATMENT OF BENIGN PROSTATIC

HYPERPLASIA

(iii) NUMBER OF SEQUENCES: 194 (iv) CORRESPONDENCE

ADDRESS:

ADDRESSEE: DAVID A. MUTHARD STREET: 126 E. Lincoln Avenue, P.O. BOX 2000 CITY: RAHWAY STATE: NEW JERSEY COUNTRY: U.S.A.

ZIP: 07065 COMPUTER READABLE FORM: MEDIUM TYPE: Floppy disk COMPUTER: IBM PC compatible OPERATING SYSTEM: PC-DOS/MS-DOS SOFTWARE: PatentIn Release Version #1.30 (vi) CURRENT APPLICATION

DATA:

APPLICATION

NUMBER:

FILING DATE:

CLASSIFICATION:

(viii) ATTORNEY/AGENT

INFORMATION:

NAME: Muthard, David A.

REGISTRATION NUMBER: 35,297 REFERENCE/DOCKET NUMBER: 19560 (ix) TELECOMMUNICATION

INFORMATION:

TELEPHONE: (908)594-3903 TELEFAX: (908)594-4720 INFORMATION FOR SEQ ID NO:1: SEQUENCE

CHARACTERISTICS:

LENGTH: 462 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO WO 97/14416 PCT/US96/16490 70 FRAGMENT TYPE: internal (xi) Met SEQUENCE DESCRIPTION: SEQ ID NO:i: Lys Pro Asn Ile Ile Phe Val Leu Ser 1 5 Lys Gin Ala Ala Val Pro Ser Ser Gly Giu Gin Se G11 Met Gly Gin Lys Gin Phe Pro His 10 Gly Giy Gly Gin Ser Lys Lys Leu Leu Leu Ile Leu Glu Gly Arc Ile Lys Gin Gly His Asp 145 Asn Ser Ser Glu Giu 225 Gin Tyr Thr Tyr Ser Arg Arg His 130 Gin Thr Val Tyr Thr 210 Val Gin His Asp 115 Lys Gly Giu Ser Val 195 Lys Arg Gin Leu 100 His Gly Asn Giu Gly 180 Leu Asn Glu Tyr Gly Asp Gly Ser Arg 165 Ala Gin Ser Glu Lys His 70 Asp Gly Lys Lys Pro 150 Leu Gin Thr His His 230 55 Val Leu Ser Ser Ala 135 Ser Trp Lys Giu Gin 215 Ser Asp Asn Gin Lys 120 His Gly Val1 Gly.

Giu 200 Asn Ser Ala Ala Gin 105 Gly Arg Lys His Arg 185 Leu Lys L~ys Asn Leu 90 Leu His Gb; Gly Gly 170 Lys Val G Iy Val Asp 75 His Leu Phe Thr Ile 155 Leu Sin A~la H1is His Lys His His Gin 140 Ser Ser Gly Asn Tyr 220 Thr Asp Thr Asn Arg 125 Asn Ser Lys Gly Lys 205 Gln Ser 40 Gin Gin Thr Glu Ser Lys Gly Ser Phe Gin *Thr Lys 110 Val Pro Gin Giu Ser 190 Gin Asn Leu Ser Lys Gin Val Ser Tyr Gin 175 Gin Gin Val aLeu 3Tyr Ser Arg Ser Giu Ile Gin Ser 160 Thr Ser Arg Val Pro 240 235 Ala His Gin Asp Lys 245 Leu Gin His Gly Ser 250 Lys Asp Ile Phe Ser Thr 255 WO 97/14416 PCT/US96/16490 Gin Asp Giu Leu Leu Vai Tyr Leu Asn Gin Asp Gin Gin Gin Ser 290 Vai Gin 305 Lys Aia Gin Giu Thr Giu Vai Ser 370 Lys Ser 385 275 Ser Lys Gin His Giu 355 Gin Gin Ser Asp Giy Ser 340 Arg Arg Ile Thr Giu Vai Ser 310 Lys Ser 325 Gin Lys Arg Leu Ser Ile Gin Aia 390 Giy Giu 405 Giu Gin Vai Ile His Giu 295 Gin Gin Aia His Tyr 375 Pro Ser Lys Ile 71 Asn 2 Giy i 280 Arg A Ser S Lys G Asn L 3 Tyr G 360 Ser G Asn P: Giy G Giy A 4: Giu GI .sys Asn ~65 Lrg Lys Gin His Gin Thr Lys 270 Aia Asn Lys Ile Ser Asn 285 ~rg :er in ys 45 iy in Ln Leu His Tyr Giy Giu Asn Giy Ile Ile 330 11ie Giu Thr Lys Ser 410 His Giu Tyr 315 Thr Ser Asn Giu Gin 395 Thr Gin Asp 300 Ser Ile Tyr Giy Lys 380 Giu Asn His Asp Gin Pro Gin Vali 365 Leu Pro Arg Giy Thr Ser Ser 350 Gin Vali Trp Giu Ser 430 Giu Gin 335 Ser Lys Aia His Gin 415 His Arg Giu 320 Giu Ser Asp Giy Giy 400 Asp Gly His Giu Asn Ala Lys Leu Gly Leu Leu Ser Asp 435 445 Leu Aia Gin His Leu Asn Asn Asp Arg Asn Pro 450 455 INFORMATION FOR SEQ ID NO:2: SEQUENCE CHARACTERISTICS: LENGTH: 15 amino acids TYPE: amino acid STRANDEDNESS: singie TOPOLOGY: iinear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal Leu Phe Thr 460

M

WO 97/14416 PCT/1S96/16490 -72- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: Gly Lys Gly Ile Ser Ser Gin Tyr Ser Asn Thr Glu Glu Arg Leu 1 5 10 INFORMATION FOR SEQ ID NO:3: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: Asn Lys Ile Ser Tyr Gin Ser Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:4: SEQUENCE CHARACTERISTICS: LENGTH: 19 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: Gly Glu Asn Gly Val Gin Lys Asp Val Ser Gin Arg Ser Ile Tyr Ser 1 5 10 Gin Thr Glu -73- *t.

INFORMATION FOR SEQ ID SEQUENCE

CHARACTERISTICS:

LENGTH: 19 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID Gly Glu Asn Gly Val Gln Lys Asp Val Ser Gln Ser Ser Ile Tyr Ser 1 5 10 Gln Thr Glu INFORMATION FOR SEQ ID NO:6: SEQUENCE

CHARACTERISTICS:

LENGTH: 25 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: Gly Arg Lys Ala Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr Glu Glu 1 5 10 Arg Arg Leu His Tyr Gly Glu Asn Gly 20 7 /L 4 -x k1

CY

II11111111 -74- INFORMATION FOR SEQ ID NO:7: SEQUENCE

CHARACTERISTICS:

LENGTH: 8 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: o' Ser Tyr Gln Ser Ser Ser Thr Glu S* 1 INFORMATION FOR SEQ ID NO:8: SEQUENCE

CHARACTERISTICS:

LENGTH: 9 amino acids So TYPE: amino acid o STRANDEDNESS: single S(D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

e (iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: Ile Ser Tyr Gln Ser Ser Ser Thr Glu 1 INFORMATION FOR SEQ ID NO:9: SEQUENCE

CHARACTERISTICS:

LENGTH: 10 amino acids TYPE: amino acid A STRANDEDNESS: single TOPOLOGY: linear ",ic -y, (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: Lys Ile Ser Tyr Gin Ser Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal

V

'K 7 x (xi) SEQUENCE DESCRIPTION: SEQ ID Asn Lys Ile Ser Tyr Gin Ser Ser Ser Thr 1 5 INFORMATION FOR SEQ ID NO:11: SEQUENCE CHARACTERISTICS: LENGTH: 12 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal -76- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11: Ala Asn Lys Ile Ser Tyr Gin Ser Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:12: SEQUENCE

CHARACTERISTICS:

LENGTH: 8 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal S (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: Ala Gly Pro Thr Gly Ala Ser Ala 1 INFORMATION FOR SEQ ID NO:13: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: s Asn Lys Ile Ser Tyr Gin Ser 1 'KAP 2: -77- INFORMATION FOR SEQ ID NO:14: SEQUENCE

CHARACTERISTICS:

LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal a.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: Lys Ile Ser Tyr Gln Ser 1 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 19 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (ix) FEATURE: NAME/KEY: Peptide LOCATION: 12 OTHER INFORMATION: /note= "any natural amino acid" (xi) SEQUENCE DESCRIPTION: SEQ ID Gly Glu Asn Gly Val Gln Lys Asp Val Ser Gln Xaa Ser Ile Tyr Ser 1 5 10 Gln Thr Glu ti

A;

N,

INFORMATION FOR SEQ ID NO:16: -78- 9*

S

S Sn

S

SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: Asn Lys Ile Ser Tyr Gin Ser Ser 1 INFORMATION FOR SEQ ID NO:17: SEQUENCE

CHARACTERISTICS:

LENGTH: 9 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: Asn Lys Ile Ser Tyr Gin Ser Ser Ser 1 INFORMATION FOR SEQ ID NO:18: SEQUENCE

CHARACTERISTICS:

LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide Vi.\ A3 1

O,

-79-

S

S.

*S

S S *5 *5.

S

(iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: Ala Asn Lys Ile Ser Tyr Gin Ser Ser Ser 1 5 INFORMATION FOR SEQ ID NO:19: SEQUENCE

CHARACTERISTICS:

LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: Lys Ile Ser Tyr Gln Ser Ser Ser Thr Glu INFORMATION FOR SEQ ID SEQUENCE

CHARACTERISTICS:

LENGTH: 17 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal i/ iil1i

I

'Ilb (xi) SEQUENCE DESCRIPTION: SEQ ID Gln Leu Asp Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr His Gln Ser Ser INFORMATION FOR SEQ ID NO:21: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear I (ii) MOLECULE TYPE: peptide iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

S(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21: Asn Arg Ile Ser Tyr Gin Ser S1 INFORMATION FOR SEQ ID NO:22: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22: Asn Lys Val Ser Tyr Gin Ser -81 INFORMATION FOR SEQ ID NO:23: SEQUENCE CHARACTERISTICS: LENGTH: 8 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal 0* (xi) Asn 1 SEQUENCE DESCRIPTION: SEQ ID NO:23: Lys Met Ser Tyr Gln Ser Ser INFORMATION FOR SEQ ID NO:24: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24: Asn Lys Leu Ser Tyr Gln Ser Ser 1 INFORMATION FOR SEQ ID SEQUENCE

CHARACTERISTICS:

LENGTH: 9 amino acids TYPE: amino acid STRANDEDNESS: single v Xf f -82so *to.

00.0 6 4.

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID Asn Lys Ile Thr Tyr Gln Ser Ser Ser 1 INFORMATION FOR SEQ ID NO:26: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26: Asn Lys Ile Ser Phe Gln Ser Ser Ser 1 INFORMATION FOR SEQ ID NO:27: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

0 -83- FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27: Asn Lys Ile Ser Trp Gin Ser Ser Ser Thr 1 5 INFORMATION FOR SEQ ID NO:28: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal a (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28: Asn Lys Ile Ser Tyr Asn Ser Ser Ser Thr 1 5 9" INFORMATION FOR SEQ ID NO:29: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29: Asn Lys Ile Ser Tyr Gin Thr Ser Ser Thr 1 5 INFORMATION FOR SEQ ID i I

VTO^''

-84-

S

ar cc a

C

9 *a S S.5 9 a

S

a..

0 SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID Asn Lys Ile Ser Tyr Gin Ser 1 INFORMATION FOR SEQ ID NO:31: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31: Gin Lys Ile Ser Tyr Gin Ser Ser 1 INFORMATION FOR SEQ ID NO:32: SEQUENCE

CHARACTERISTICS:

LENGTH: 9 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide ;;VT o' (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal

A.

y- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32: Asn Arg Ile Thr Tyr Gin Ser Ser Ser 1 INFORMATION FOR SEQ ID NO:33: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33: Asn Arg Ile Ser Phe Gin Ser Ser Ser Thr 1 5 INFORMATION FOR SEQ ID NO:34: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal -86- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34: Gin Lys Ile Ser Tyr Gin Thr Ser Ser Thr 1 5 INFORMATION FOR SEQ ID SEQUENCE

CHARACTERISTICS:

LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide o*o* (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID Asn Arg Ile Ser Trp Gin Ser Ser Ser Thr *10 INFORMATION FOR SEQ ID NO:36: SEQUENCE

CHARACTERISTICS:

LENGTH: 10 amino acids S* TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36: Asn Arg Ile Ser Tyr Gin Thr Ser Ser Thr 1 5 INFORMATION FOR SEQ ID NO:37: i/ j i -87- SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37: SAsn Lys Ile Thr Tyr Gin Thr Ser Ser Thr S1 5 INFORMATION FOR SEQ ID NO:38: SEQUENCE

CHARACTERISTICS:

LENGTH: 10 amino acids S. TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

o* (iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38: Asn Lys Leu Ser Tyr Gin Thr Ser Ser Thr 1 5 INFORMATION FOR SEQ ID NO:39: SEQUENCE

CHARACTERISTICS:

LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide -88- (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39: Gin Lys Leu Ser Tyr Gin Ser Ser Ser Thr 1 5 INFORMATION FOR SEQ ID SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal *aa.

.o S(xi) SEQUENCE DESCRIPTION: SEQ ID Asn Arg Leu Ser Tyr Gin Thr Ser Ser Thr 1 5 INFORMATION FOR SEQ ID NO:41: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal rT -89- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41: Asn Lys Val Ser Phe Gin Ser Ser Ser Thr 1 5 INFORMATION FOR SEQ ID NO:42: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42: sn Arg Val Ser Trp Gin Ser Ser Ser Thr *1 5 INFORMATION FOR SEQ ID NO:43: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43: Gin Lys Val Ser Tyr Gin Ser Ser Ser Thr INFORMATION FOR SEQ ID NO:44: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44: S* Asn Lys Ile Ser Tyr Gin Ser Ser Ser Thr 1 5 i. INFORMATION FOR SEQ ID a SEQUENCE

CHARACTERISTICS:

LENGTH: 19 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID Gly Glu Gin Gly Val Gin Lys Asp Val Ser Gin Ser Ser Ile Tyr Ser 1 5 10 Gin Thr Glu INFORMATION FOR SEQ ID NO:46: SEQUENCE

CHARACTERISTICS:

LENGTH: 15 amino acids TYPE: amino acid A STRANDEDNESS: single -91 TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46: Gly Lys Gly Ile Ser Ser Gln Tyr Ser Asn Thr Asp Glu Arg Leu 10 INFORMATION FOR SEQ ID NO:47: S(i) SEQUENCE CHARACTERISTICS: LENGTH: 19 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO FRAGMENT TYPE: internal *4a.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:47: Gly Glu Asn Gly Leu Gln Lys Asp Val Ser Gin Ser Ser Ile Tyr Ser 1 5 10 Gin Thr Glu INFORMATION FOR SEQ ID NO:48: SEQUENCE

CHARACTERISTICS:

NO

-92- (iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48: Gly Glu Asn Gly Val Asn Lys Asp Val Ser Gin Ser Ser Ile Tyr Ser 1 5 10 15 1 0 Gin Thr Glu INFORMATION FOR SEQ ID NO:49: SEQUENCE CHARACTERISTICS: LENGTH: 19 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49: Gly Glu Asn Gly Val Gln Arg Asp Val Ser Gin Arg Ser Ile Tyr Ser 1 5 10 Gin Thr Glu INFORMATION FOR SEQ ID SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

-r -93-

C

S

C

ft ft 5f *aft.

t f a.

*5 t S f

*SC*

f

C..

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID Gly Glu Asn Gly Val Gin Lys Asp Val Ser Gin Lys Ser Ile Tyr Ser 1 5 10 Gin Thr Glu INFORMATION FOR SEQ ID NO:51: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51: Gly Glu Asn Gly Val Gln Lys Asp Leu Ser Gln Thr Ser Ile Tyr Ser 1 5 10 Gin Thr Glu INFORMATION FOR SEQ ID NO:52: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal h -94- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52: Gly Glu Asn Gly Val Gin Lys Asp Val Ser Gin Ser Ser Ile Phe Ser 1 5 10 Gin Thr Glu INFORMATION FOR SEQ ID NO:53: SEQUENCE

CHARACTERISTICS:

NO

S* (iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53: Gly Glu Asn Gly Val Gln Lys Asp Met Ser Gln Ser Ser Ile Tyr Thr Gin Thr Glu INFORMATION FOR SEQ ID NO:54:

S

SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54: Gly Glu Asn Gly Val Gln Lys Asp Val Ser Gln Arg Ser Ile Tyr Thr 1 5 10 iJ, Gln Thr Glu INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 19 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID Gly Glu Asn Gly Val Gin Lys Asp Val Ser Gin Ser Ser Ile Tyr Ser S. 10 Gin Ser Glu o e INFORMATION FOR SEQ ID NO:56: SEQUENCE

CHARACTERISTICS:

LENGTH: 19 amino acids S* TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56: Gly Glu Asn Gly Val Gln Lys Asp Val Ser Gln Arg Ser Ile Tyr Ser 1 5 10 Asn Thr Glu -96- INFORMATION FOR SEQ ID NO:57: SEQUENCE

CHARACTERISTICS:

LENGTH: 15 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57: SGly Lys Ala Ile Ser Ser Gin Tyr Ser Asn Thr Glu Glu Arg Leu 10 INFORMATION FOR SEQ ID NO:58: SEQUENCE CHARACTERISTICS: LENGTH: 15 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58: Gly Lys Gly Ile Ser Ser Gin Tyr Ser Asn Ser Glu Glu Arg Leu 10 INFORMATION FOR SEQ ID NO:59: SEQUENCE

CHARACTERISTICS:

LENGTH: 15 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear 1 -97- (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59: Gly Arg Gly Ile Ser Ser Gln Tyr Ser Asn Thr Glu Glu Arg Leu 1 5 INFORMATION FOR SEQ ID SEQUENCE

CHARACTERISTICS:

LENGTH: 15 amino acids TYPE: amino acid STRANDEDNESS: single S" TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

U

ANTI-SENSE: NO FRAGMENT TYPE: internal SEQUENCE DESCRIPTION: SEQ ID Gly Lys Gly Ile Thr Ser Gln Tyr Ser Asn Thr Glu Glu Arg Leu INFORMATION FOR SEQ ID NO:61: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal i' -98- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61: Gly Lys Gly Ile Ser Thr Gln Tyr Ser Asn Thr Glu Glu Arg Leu 10 INFORMATION FOR SEQ ID NO:62: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62: SGly Lys Gly Ile Ser Ser Asn Tyr Ser Asn Thr Glu Glu Arg Leu *o 101 1 0 INFORMATION FOR SEQ ID NO:63: a SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63: Ala Lys Gly Ile Ser Ser Gln Tyr Ser Asn Thr Glu Glu Arg Leu 10 INFORMATION FOR SEQ ID NO:64: I r -99- SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64: Gly Lys Gly Ile Ser Ser Gln Phe Ser Asn Thr Glu Glu Arg Leu 1 0 1 INFORMATION FOR SEQ ID *0* SEQUENCE

CHARACTERISTICS:

LENGTH: 15 amino acids TYPE: amino acid STRANDEDNESS: single :0 TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

o FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID Gly Lys Gly Ile Ser Ser Gln Tyr Thr Asn Ser Glu Glu Arg Leu 1 5 10 1 0 INFORMATION FOR SEQ ID NO:66: SEQUENCE

CHARACTERISTICS:

LENGTH: 15 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear ,V (ii) MOLECULE TYPE: peptide i j l -100- (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66: Gly Lys Gly Ile Ser Ser Gin Tyr Ser Asn Ser Glu Glu Arg Leu 1 5 10 1 0 INFORMATION FOR SEQ ID NO:67: SEQUENCE

CHARACTERISTICS:

LENGTH: 25 amino acids TYPE: amino acid S STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67: SSer Gln Lys Ala Asn Lys Ile Ser Tyr Gin Ser Ser Ser Thr Glu Glu Arg Arg Leu His Tyr Gly Glu Asn Gly INFORMATION FOR SEQ ID NO:68: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal -101 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68: Ile Ser Tyr Gin Ser Ser Ser Thr 1 INFORMATION FOR SEQ ID NO:69: SEQUENCE

CHARACTERISTICS:

LENGTH: 9 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide J (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal *0 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69: Ala Asn Lys Ile Ser Tyr Gin Ser Ser 1 INFORMATION FOR SEQ ID SEQUENCE

CHARACTERISTICS:

LENGTH: 12 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID Ala Asn Lys Ile Ser Tyr Gin Ser Ser Ser Thr Leu 1 5 t \i 'T y -102- INFORMATION FOR SEQ ID NO:71: SEQUENCE CHARACTERISTICS: LENGTH: 12 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71: Ala Asn Gly Ile Ser Tyr Gin Ser Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:72: SEQUENCE CHARACTERISTICS: LENGTH: 12 amino acids i* TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72: Ala Asn Pro Ile Ser Tyr Gln Ser Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:73: SEQUENCE

CHARACTERISTICS:

LENGTH: 12 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear -103- 0 0 0 (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:73: Ala Asn Lys Ile Ser Tyr Gln Ser Ala Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:74: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74: Ala Asn Lys Ile Ser Tyr Gln Ser Ser Lys Thr Glu

I

INFORMATION FOR SEQ ID SEQUENCE

CHARACTERISTICS:

LENGTH: 11 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal ;v -104- (xi) SEQUENCE DESCRIPTION: SEQ ID Ala Asn Lys Ile Ser Tyr Gin Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:76: SEQUENCE

CHARACTERISTICS:

LENGTH: 12 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO ANTI-SENSE: NO FRAGMENT TYPE: internal (ix) FEATURE: NAME/KEY: Peptide LOCATION: OTHER INFORMATION: /label= d-serine /note "unnatural configuration of the amino acid" 0 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:76: *ooo Ala Asn Lys Ile Ser Tyr Gin Ser Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:77: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal 0 9'' -105- (ix) FEATURE: NAME/KEY: Peptide LOCATION: 4 OTHER INFORMATION: /label= d-isoleucine /note= "unnatural amino acid stereochemical configuration" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:77: Ala Asn Lys Ile Ser Tyr Gin Ser Ser Ser Thr Glu 1 5 a a a a..

a a a INFORMATION FOR SEQ ID NO:78: SEQUENCE CHARACTERISTICS: LENGTH: 12 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:78: Ala Asn Lys Ile Ser Tyr Gin Ser Ser Gin Thr Glu 1 5 INFORMATION FOR SEQ ID NO:79: SEQUENCE CHARACTERISTICS: LENGTH: 12 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal

C'

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:79: -106- Ala Asn Lys Ile Ser Tyr Gin Ser Ala Lys Thr Glu 1 5 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 12 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal S* (ix) FEATURE: NAME/KEY: Peptide LOCATION: 3 OTHER INFORMATION: /label= d-lysine /note= "unnatural amino acid stereochemical configuration" (xi) SEQUENCE DESCRIPTION: SEQ ID :Ala Asn Lys Ile Ser Tyr Gin Ser Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:81: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:81: Ala Asn Lys Ile Ser Tyr Gin Ser Thr Glu 1 5 -107- INFORMATION FOR SEQ ID NO:82: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal i (xi) SEQUENCE DESCRIPTION: SEQ ID NO:82: Ala Asn Lys Ser Tyr Gin Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:83: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids 9 TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:83: Ala Asn Lys Ile Tyr Gln Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:84: SEQUENCE

CHARACTERISTICS:

LENGTH: 12 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear IIIi -108- 0 0 0 0 *000 0 0* (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:84: Ala Asn Lys Ala Ser Tyr Gin Ser Ala Ser Thr Glu 1 5 INFORMATION FOR SEQ ID SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID Ala Asn Glu Ile Ser Tyr Gin Ser Ala Ser Thr Glu INFORMATION FOR SEQ ID NO:86: SEQUENCE

CHARACTERISTICS:

LENGTH:, 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal i 0 0

I

-109- (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:86: Lys Ile Ser Tyr Gin Ser Ser 1 INFORMATION FOR SEQ ID NO:87: SEQUENCE

CHARACTERISTICS:

NO

S(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:87: SSer Tyr Gin Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:88: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:88: Ser Tyr Gln Ser Ser Thr Leu 1 A 1 d' -110-

S.

S

*5*

*S.

S

*r S

S

INFORMATION FOR SEQ ID NO:89: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:89: Ala Ser Tyr Gln Ser Ser Thr Glu 1 INFORMATION FOR SEQ ID SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID Glu Ile Ser Tyr Gln Ser Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:91: SEQUENCE

CHARACTERISTICS:

LENGTH: 12 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear

K

i IC 111 (ii) (iii) (iv) (v) MOLECULE TYPE: peptide HYPOTHETICAL:

NO

ANTI-SENSE:

NO

FRAGMENT TYPE: internal p p.

p p p 9**p pp.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:91: Ala Asn Glu Ile Ser Tyr Gin Ser Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:92: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:92: Ala Asn Lys Ile Ser Tyr Tyr Ser Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:93: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal -112- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:93: Ala Asn Lys Ile Ser Tyr Tyr Ser Ala Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:94: SEQUENCE

CHARACTERISTICS:

NO

S(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal o a ooeo a a.

*aa.

a a ao a a o• a a a ***ft (xi) SEQUENCE DESCRIPTION: SEQ ID NO:94: Ala Ser Tyr Gin Ser Ser Leu 1 INFORMATION FOR SEQ ID SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID Ala Asn Ser Tyr Gin Ser Ser Ser Thr Glu 1 5 2-

,A

'Y@

-113- INFORMATION FOR SEQ ID NO:96: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal

S

S.

S

p.

p

S..

9.

B

S p 9*

B

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:96: Ala Ser Tyr Gln Ser Ser Ser Thr Glu 1 INFORMATION FOR SEQ ID NO:97: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:97: Ser Tyr Gln Ser Ser Ser Thr Glu 1 INFORMATION FOR SEQ ID NO:98: SEQUENCE

CHARACTERISTICS:

LENGTH: 12 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear i i ~i i: t\ -114- (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:98: Ala Asn Lys Ala Ser Tyr Gin Ser Ala Ser Thr Cys 1 5 INFORMATION FOR SEQ ID NO:99: SEQUENCE CHARACTERISTICS: LENGTH: 5 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

*OS*

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:99: Gin Ser Ser Thr Glu 1 INFORMATION FOR SEQ ID NO:100: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal J -115- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:100: Tyr Gin Ser Ser Thr Glu 1 INFORMATION FOR SEQ ID NO:101: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

0 (iv) ANTI-SENSE: NO o* FRAGMENT TYPE: internal 0.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:101: S" Ser Gin Ser Ser Thr Glu 1 INFORMATION FOR SEQ ID NO:102: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:102: Ala Asn Lys Ile Ser Gin Ser Ser Thr Glu 1 5 i: t D -116- INFORMATION FOR SEQ ID NO:103: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (ix) FEATURE: NAME/KEY: Peptide LOCATION: 3 OTHER INFORMATION: /label= unnatural /note= "ornithine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:103: Ala Asn Xaa Ile Ser Tyr Gln Ser Ser Thr Glu 1 5 go 0o INFORMATION FOR SEQ ID NO:104: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: /label= unnatural /note= "3, 4 -dichlorophenalanine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:104: o Ser Xaa Gln Ser Ser Thr Glu I) C -117- INFORMATION FOR SEQ ID NO:105: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: /label= unnatural /note= 3 -pyridinyl)alanine" *r a. *a *r a.0 a.

a a..

a.

a a a a.

(xi) Ser 1 SEQUENCE DESCRIPTION: SEQ ID NO:105: Xaa Gln Ser Ser Thr Glu INFORMATION FOR SEQ ID NO:106: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:106: Ser Lys Gln Ser Ser Thr Glu 1 INFORMATION FOR SEQ ID NO:107: SEQUENCE

CHARACTERISTICS:

-118- LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:107: Ser Tyr Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:108: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear Vo (ii) MOLECULE TYPE: protein Ve

V

(iii) HYPOTHETICAL:

NO

oo** (iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /label= unnatural /note= "epsilon aminocaproic acid" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:108: Xaa Tyr Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:109: SEQUENCE

CHARACTERISTICS:

LENGTH: 11 amino acids TYPE: amino acid STRANDEDNESS: single V T w -119- TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (ix) FEATURE: NAME/KEY: Peptide LOCATION: 4 OTHER INFORMATION: /label= unnatural /note= "N-methylisoleucine"

S

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:109: Ala Asn Lys Xaa Ser Tyr Gln Ser Ser Thr Glu 1 5 INFORMATION FOR SEQ ID NO:110: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear S.

(iL) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:110: Ser Tyr Gln Ser Ser Thr Glu 1 INFORMATION FOR SEQ ID NO:111: SEQUENCE

CHARACTERISTICS:

LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide

I

-120a a *a a.

a.

(iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:111: Tyr Gln Ser Ser Thr Glu 1 INFORMATION FOR SEQ ID NO:112: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:112: Ser Tyr Lys Ser Ser Thr Glu *9a.

a *aa.

INFORMATION FOR SEQ ID NO:113: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal r -121 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:113: Ser Tyr Tyr Ser Ser Thr Glu 1 INFORMATION FOR SEQ ID NO:114: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal 9*

S

S.

S

9 9 9**c 9.

9 9* 55.9 S 95r

S

s *9 9 9 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:114: Ser Tyr Gin Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:115: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) Ser 1 SEQUENCE DESCRIPTION: SEQ ID NO:115: Tyr Gin Ser Ser Leu INFORMATION FOR SEQ ID NO:116: SEQUENCE CHARACTERISTICS: i i A i

T

-122- (ii) (iii) (iv) (v) LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear MOLECULE TYPE: peptide HYPOTHETICAL:

NO

ANTI-SENSE: NO FRAGMENT TYPE: internal (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /label: /note= "2, 3 -diaminopropionic acid" *r

C

unnatural (xi) Xaa 1 SEQUENCE DESCRIPTION: SEQ ID NO:116: Tyr Gln Ser Ser Ser Leu be C C

L.

C

be a.

C

bCC...

C

INFORMATION FOR SEQ ID NO:117: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid STRANDEDNESS: single (D).TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:117: Ala Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr 1 5 INFORMATION FOR SEQ ID NO:118: SEQUENCE CHARACTERISTICS: LENGTH: 12 amino acids TYPE: amino acid STRANDEDNESS: single 1'-d;r o., -123- TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:118: Ala Asn Lys Ala Ser Tyr Gln Ser Ala Ser Thr Leu 1 5 INFORMATION FOR SEQ ID NO:119: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal

U.

U

U.

U

S I Ut.

S..

U U

U

S

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:119: Ala Asn Lys Ala Ser Tyr Gln Ser Ala Ser Leu 1 5 INFORMATION FOR SEQ ID NO:120: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

_-0 -124- FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:120: Ala Asn Lys Ala Ser Tyr Gin Ser Ser Ser Leu 1 5 INFORMATION FOR SEQ ID NO:121: SEQUENCE

CHARACTERISTICS:

LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

S(iv) ANTI-SENSE:

NO

S(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:121: Ala Asn Lys Ala Ser Tyr Gin Ser Ser Leu 1 5 INFORMATION FOR SEQ ID NO:122: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /label= d-leucine /note= "unnatural amino acid stereochemical configuration \e i ,iij -125- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:122: Ser Tyr Gin Ser Ser Thr Leu 1 INFORMATION FOR SEQ ID NO:123: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal 9 9* 9 9 9 9999 99..

9 9 *999 9 9e.

99 9 9 99 p 9 9999 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:123: Ala Asn Lys Ala Ser Tyr Ala Ser Ser Ser Leu 1 5 INFORMATION FOR SEQ ID NO:124: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:124: Lys Tyr Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:125: SEQUENCE

CHARACTERISTICS:

Y:

i: 2 ~7 i^ -126- LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:125: Ser Tyr Gln Ser Ser Lys Leu 1 9@ INFORMATION FOR SEQ ID NO:126: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide t (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /label= d-leucine /note= "unnatural amino acid stereochemical configuration" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:126: Ser Tyr Gln Ser Ser Lys Leu 1 INFORMATION FOR SEQ ID NO:127: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single -127- TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:127: Asn Lys Ile Ser Tyr Tyr Ser 1 INFORMATION FOR SEQ ID NO:128: 9e SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO S(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:128: Asn Lys Ala Ser Tyr Gin Ser 1 INFORMATION FOR SEQ ID NO:129: SEQUENCE CHARACTERISTICS: LENGTH: 5 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO -128- FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:129: Ser Tyr Gin Ser Ser 1 INFORMATION FOR SEQ ID NO:130: SEQUENCE CHARACTERISTICS: LENGTH: 8 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO FRAGMENT TYPE: internal o. (xi) SEQUENCE DESCRIPTION: SEQ ID NO:130: Asn Lys Ile Ser Tyr Gin Ser Ala 1 INFORMATION FOR SEQ ID NO:131: SEQUENCE CHARACTERISTICS: LENGTH: 8 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:131: Ala Asn Lys Ile Ser Tyr Tyr Ser (d -129- 1 INFORMATION FOR SEQ ID NO:132: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal S* (xi) SEQUENCE DESCRIPTION: SEQ ID NO:132: Ala Asn Lys Ala Ser Tyr Gln Ser 1 INFORMATION FOR SEQ ID NO:133: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:133: Ser Tyr Gln Ser Ser Thr 1 INFORMATION FOR SEQ ID NO:134: SEQUENCE

CHARACTERISTICS:

LENGTH: 6 amino acids TYPE: amino acid S,.'ir STRANDEDNESS: single

N<

k ,i -130- 0* 4**c I I

I.

I

II..

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:134: Ser Tyr Gin Ser Ser Ser 1 INFORMATION FOR SEQ ID NO:135: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:135: Ser Tyr Gln Ser Ser Leu INFORMATION FOR SEQ ID NO:136: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

i -131 FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:136: Ala Asn Lys Ile Ser Tyr Gin Ser Ala 1 INFORMATION FOR SEQ ID NO:137: SEQUENCE CHARACTERISTICS: LENGTH: 9 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide S* (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE:

NO

S..

FRAGMENT TYPE: internal S. (xi) SEQUENCE DESCRIPTION: SEQ ID NO:137:

S.

Ala Asn Lys Ile Ser Tyr Tyr Ser Ser 1 INFORMATION FOR SEQ ID NO:138: **too* SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:138: Ala Asn Lys Ile Ser Tyr Tyr Ser Ala -132- 1 INFORMATION FOR SEQ ID NO:139: SEQUENCE

CHARACTERISTICS:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal e S(xi) SEQUENCE DESCRIPTION: SEQ ID NO:139: Ala Asn Lys Ala Ser Tyr Gln Ser Ala INFORMATION FOR SEQ ID NO:140: SEQUENCE CHARACTERISTICS: to LENGTH: 5 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide S* (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:140: Lys Tyr Gln Ser Ser 1 INFORMATION FOR SEQ ID NO:141: SEQUENCE

CHARACTERISTICS:

LENGTH: 5 amino acids TYPE: amino acid S STRANDEDNESS: single 1 -133- 0 05 0 0 0

S

0 9* Sb 61 0 TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /label= homoarginine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:141: Xaa Tyr Gin Ser Ser 1 INFORMATION FOR SEQ ID NO:142: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:142: Lys Tyr Gin Ser Ser Ser 1 INFORMATION FOR SEQ ID NO:143: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide c j( i -134- (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE:

NO

FRAGMENT TYPE: internal (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /label= homoarginine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:143: Xaa Tyr Gin Ser Ser Ser eo 4* 5* *a aa o INFORMATION FOR SEQ ID NO:144: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:144: Ser Tyr Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:145: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL:

NO

(iv) ANTI-SENSE: NO -135- FRAGMENT TYPE: internal (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /label= homoarginine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:145: Xaa Tyr Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:146: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear S(ii) MOLECULE TYPE: peptide .r (iii) HYPOTHETICAL: NO ANTI-SENSE: NO FRAGMENT TYPE: internal a (ix) FEATURE: NAME/KEY: Peptide LOCATION: 6 OTHER INFORMATION: /label= norleucine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:146: Lys Tyr Gln Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:147: SEQUENCE CHARACTERISTICS: LENGTH: 5 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 -136- OTHER INFORMATION: /product= "cyclohexylalanine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:147: Xaa Xaa Gln Ser Leu a a a a INFORMATION FOR SEQ ID NO:148: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: (ix) FEATURE: NAME/KEY: Peptide LOCATION: 6 OTHER INFORMATION: /product= "homoarginine" /product= "homotyrosine" /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:148: Xaa Xaa Gln Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:149: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single -137- TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: (ix) FEATURE: NAME/KEY: Peptide LOCATION: 6 OTHER INFORMATION: /product= "homoarginine" /product= "cyclohexylhomoalanine" /product= "norleucine"

S

SOS.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:149: Xaa Xaa Gin Ser Ser Leu

S

*55*

S

INFORMATION FOR SEQ ID NO:150: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: OTHER INFORMATION: /product= "cyclohexylalanine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:150: Ala Asn Lys Ala Ser Tyr Gin Ser Ser Xaa INFORMATION FOR SEQ ID NO:151: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear -138- (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:151: Xaa Tyr Gin Ser Ser Pro 1 INFORMATION FOR SEQ ID NO:152: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide *r a a a (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:152: Xaa Tyr Gin Ser Ser His 1 INFORMATION FOR SEQ ID NO:153: SEQUENCE CHARACTERISTICS: LENGTH: 5 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:153: Xaa Tyr Gin Ser Asn U.

N;

-139- 1 INFORMATION FOR SEQ ID NO:154: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:154: Xaa Tyr Gln Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:155: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "4-aminomethylphenylalanine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:155: Xaa Tyr Gln Ser Ser Ser Leu 1 -140- INFORMATION FOR SEQ ID NO:156: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix)

FEATURE:

NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:156: Xaa Tyr Gln Ser Ser Ser Leu INFORMATION FOR SEQ ID NO:157: SEQUENCE

CHARACTERISTICS:

LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: OTHER INFORMATION: /product= "cyclohexylalanine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:157: Ala Asn Lys Ala Lys Tyr Gln Ser Ser Xaa 1 510 INFORMATION FOR SEQ ID NO:158: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide -141 (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "2(4,6-dimethylpyrimidine)lysine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:158: Xaa Tyr Gln Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:159: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide 9 S: (ix) FEATURE: *9 NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "N'-(2-imidazolyl)lysine" o*99 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:159: Xaa Tyr Gln Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:160: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: OTHER INFORMATION: /product= "homoarginine" -142- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:160: Ala Asp Lys Ala Xaa Tyr Gin Ser Ser Leu 1 5 INFORMATION FOR SEQ ID NO:161: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 S(D) OTHER INFORMATION: /product= "(4-aminocyclohexyl)alanine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 6 OTHER INFORMATION: /product= "norleucine" e (xi) SEQUENCE DESCRIPTION: SEQ ID NO:161: p Xaa Tyr Gin Ser Ser Leu 1 p INFORMATION FOR SEQ ID NO:162: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "N'-(2-imidazolylyl)sine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /product= "norleucine" li 1 -143- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:162: Xaa Tyr Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:163: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: /product= "cyclohexylalanine" (ix) FEATURE: NAME/KEY: Peptide i LOCATION: 7 OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:163: Xaa Xaa Gin Ser Ser Ser Leu 1 S(2) INFORMATION FOR SEQ ID NO:164: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 -144- OTHER INFORMATION: /product= "homoarginine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:164: Xaa Tyr Gin Ser Ser Ser Xaa 1 INFORMATION FOR SEQ ID NO:165: SEQUENCE

CHARACTERISTICS:

LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide Sa LOCATION: OTHER INFORMATION: /product= 2 -imidazolyl)lysine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:165: Ala Asn Lys Ala Xaa Tyr Gin Ser Ser Leu 1 5 i INFORMATION FOR SEQ ID NO:166: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: /product= "3-iodotyrosine" -145- (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:166: Xaa Xaa Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:167: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide a a..

a.

a (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: "O-dimethylphosphotyrosine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /product= "homoarginine" /product= /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:167: Xaa Xaa Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:168: SEQUENCE

CHARACTERISTICS:

LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: double TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: I -146- NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:168: Xaa Tyr Gin Ser Ser Asp 1 INFORMATION FOR SEQ ID NO:169: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (ix) FEATURE: NAME/KEY: Peptide o. LOCATION: 2 S* OTHER INFORMATION: /product= "O-methyltyrosine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /product= "norleucine" 9 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:169: Xaa Xaa Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:170: SEQUENCE

CHARACTERISTICS:

LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: -147- OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:170: Ala Asn Lys Ala Lys Tyr Gin Ser Ser Leu 1 5 INFORMATION FOR SEQ ID NO:171: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:

S

NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" SS (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: /product "cyclohexylalanine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:171: Xaa Xaa Gln Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:172: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= 2 -imidazolyl)lysine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: /product= "cyclohexylalanine" -148- (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:172: Xaa Xaa Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:173: SEQUENCE

CHARACTERISTICS:

LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide S.(ix)

FEATURE:

NAME/KEY: Peptide Vo LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: /product= "cyclohexylalanine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:173: 6* Xaa Xaa Gin Ser Ser Ser 1 INFORMATION FOR SEQ ID NO:174: SEQUENCE

CHARACTERISTICS:

LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (ix) FEATURE: NAME/KEY: Peptide -149- LOCATION: 2 OTHER INFORMATION: /product= "cyclohexylalanine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 6 OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:174: Xaa Xaa Gln Ser Ser Leu

S

*e INFORMATION FOR SEQ ID NO:175: SEQUENCE

CHARACTERISTICS:

S

S.

S

(ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: (ix) FEATURE: NAME/KEY: Peptide LOCATION: 6 OTHER INFORMATION: /product= "homoarginine" /product= "cyclohexylalanine" /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:175: Xaa Xaa Gln Ser Pro Leu 1 INFORMATION FOR SEQ ID NO:176: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide -150- (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: /product= 3 -fluorotyrosine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 i OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:176: Xaa Xaa Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:177: SEQUENCE CHARACTERISTICS: LENGTH: 5 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:177: Xaa Tyr Gln Ser Pro 1 INFORMATION FOR SEQ ID NO:178: SEQUENCE

CHARACTERISTICS:

LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: -151- NAME/KEY: Peptide LOCATION: 6 OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:178: Lys Tyr Gin Ser Lys Leu INFORMATION FOR SEQ ID NO:179: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide

S

S.

S

0

S

*SS*

S

*SSS

S

S S

S.

S

*.SS

S

(ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /product= "homoarginine" /product= "4-aminophenylalanine" /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:179: Xaa Xaa Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:180: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 -152- OTHER INFORMATION: /product= "homoarginine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: /product= "7-HO-tetrahydroisoquinoline CO2H" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:180: Xaa Xaa Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:181: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single S: TOPOLOGY: linear o: (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 *a-:ai OTHER INFORMATION: /product= "ornithine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:181: Xaa Tyr Gln Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:182: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide -153- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:182: Lys Ala Ala Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:183: SEQUENCE

CHARACTERISTICS:

'I

(ix) FEATURE: NAME/KEY: Peptide LOCATION: 7 OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:183: o Lys Tyr Gln Ser Ser Ser Leu *1 INFORMATION FOR SEQ ID NO:184: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:184: Leu Asn Lys Ala Ser Tyr Gln Ser Ser Leu 1 5 INFORMATION FOR SEQ ID NO:185: SEQUENCE

CHARACTERISTICS:

LENGTH: 5 amino acids TYPE: amino acid STRANDEDNESS: single S TOPOLOGY: linear -154- (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 2 OTHER INFORMATION: /product= "cyclohexylalanine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:185: Xaa Xaa Gln Ser Ser 1 INFORMATION FOR SEQ ID NO:186: S. SEQUENCE CHARACTERISTICS: LENGTH: 4 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide a (xi) SEQUENCE DESCRIPTION: SEQ ID NO:186: Tyr Gln Ser Ser 1 INFORMATION FOR SEQ ID NO:187: SEQUENCE

CHARACTERISTICS:

LENGTH: 4 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:187 -155- Xaa Tyr Gin Ser 1 INFORMATION FOR SEQ ID NO:188: SEQUENCE CHARACTERISTICS: LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "homoarginine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:188: Xaa Tyr Gin Ser Ser Ser 1 INFORMATION FOR SEQ ID NO:189: SEQUENCE CHARACTERISTICS: LENGTH: 5 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Peptide LOCATION: OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:189: Gin Ser Ser Ser Leu 1 INFORMATION FOR SEQ ID NO:190: SEQUENCE

CHARACTERISTICS:

LENGTH: 6 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide -156- (ix) FEATURE: NAME/KEY: Peptide LOCATION: 1 OTHER INFORMATION: /product= "7-HO-tetrahydro-3-isoquinoline CO2H" (ix) FEATURE: NAME/KEY: Peptide LOCATION: 6 OTHER INFORMATION: /product= "norleucine" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:190: Xaa Gln Ser Ser Ser Leu S1 o INFORMATION FOR SEQ ID NO:191: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide a a (xi) SEQUENCE DESCRIPTION: SEQ ID NO:191: o. ,Ala Asn Lys Ala Ser Tyr Ala Ser Ser Ser Leu 1 5 INFORMATION FOR SEQ ID NO:192: SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:192: Ser Tyr Gln Ser Ser Lys Leu 1 INFORMATION FOR SEQ ID NO:193: SEQUENCE

CHARACTERISTICS:

-157- LENGTH: 9 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: Peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:193: Ala Arg Lys Ala Ser Tyr Gln Ser Leu 1 INFORMATION FOR SEQ ID NO:194: OS(i) SEQUENCE

CHARACTERISTICS:

LENGTH: 7 amino acids TYPE: amino acid ()STRANDEDNESS: snl 0. TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix)

FEATURE:

AEKY Peptide A. LOCATION: 1 OTHER INFORMATION: /product= "ornithine" 0005 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:194: Xaa Tyr Gln Ser Ser Ser Leu 1C

Claims

1. A method of treating an adverse condition of the prostate which comprises administering to a mammal in need of said treatment a conjugate, said conjugate which comprises a pharmaceutical agent, effective in the treatment of said condition, attached to an oligopeptide, wherein the oligopeptide comprises a sequence of amino acids that is recognized and selectively proteolytically cleaved by free prostate specific antigen, wherein the means of attachment is directly through a covalent bond or via a linker unit, or the pharmaceutically acceptable salt thereof.

2. A method of treating benign prostatic hyperplasia which comprises administering to a mammal in need of said treatment a conjugate, said conjugate which comprises a cytotoxic agent attached to an oligopeptide, wherein the oligopeptide comprises a sequence of amino acids that is recognized and selectively proteolytically cleaved by free prostate specific antigen, wherein the means of attachment is directly through a covalent bond or via a linker unit, or the pharmaceutically acceptable salt thereof.

3. The method of treatment according to Claim 2 wherein the cytotoxic agent is a member of a class of cytotoxic agents selected from the following classes: a) anthracycline family of drugs, b) the vinca alkaloid drugs, c) the mitomycins, d) the bleomycins, e) the cytotoxic nucleosides, f) the pteridine family of drugs, g) diynenes, h) estramustine, AMENDED SHEET f9560Y ~TU9 6/16 49 0 106 Rec'd PCTI'PTO N ~OV 1997 -159- i) cyclophosphaniide, j) the podophyllotoxins, and k) the taxanes; or the pharmaceutically acceptable salt thereof.

4. The method of treatment according to Claim 2 wherein the cytotoxic agent is selected from the following cytotoxic agents: a) doxorubicin, b) carminomycin, c) daunorubicin, d) aminopterin, e) methotrexate, f) methopterin, g) dichloro-methotrexate, h) initomycin C, i) porfiromycin, j) k) 6 -mercaptopurine, 1) cytosine arabinoside, m) podophyllotoxin, n) etoposide, o) etoposide phosphate, p) melphalan, q) vinblastine, r) vincristine, s) leurosidine, t) vindesine, u) estramustine, v) cisplatin, w) cyclophosphanide, x) leurosine, and y) taxol; AMENDED. SH~EET 0 f9560Y PCT/US 96/16490 106 Rec'd PCT/PTO 7 NOV 1997 -160- or the pharmaceutically acceptable salt thereof. The method of treatment according to Claim 2 wherein the cytotoxic agent is selected from doxorubicin, vinblastine and desacetylvinblastine or a cytotoxic derivative thereof.

6. The method of treatment according to Claim 2 wherein the cytotoxic agent is selected from vinblastine and desacetylvinblastine or a cytotoxic derivative thereof.

7. The method of treatment according to Claim wherein the conjugate is of the formula I: 0 OH 0 CH2OH OH CH 3 O O OH O CH 3 0 NH OH XL oligopeptide- R wherein: oligopeptide is an oligopeptide which is specifically recognized by the free prostate specific antigen (PSA) and is capable of being proteolytically cleaved by the enzymatic activity of the free prostate specific antigen; ~ET I9560Y PCTIUS 9 6/16490 106 Rec'd PCTiPTO 17 No0 1,9 161 XL is absent or is an amino acid selected from: a) phenylalanine, b) leucine, c) valine, d) isoleucine, e) (2-naphthyl)alanine, f) cyclohexylalanine, g) diphenylalanine, h) norvaline, i) norleucine, and j) 1,2,3 4 -tetrahydroisoquinohine3carboxylic acid; R is hydrogen or 1; and R1 is ClI-C6-alkyl or aryl, or the pharmaceutically acceptable salt thereof.

8. The method of treatment according to Claim 7 wherein: oligopeptide is an oligomer that comprises an amino acid sequence selected from: a) AsnLyslleSerTyrcjlnlSer (SEQ.ID.NO.: 13),- b) LyslleSerTyrGlnlSer (SEQ.ID.No.: 14), c) Gl~us~yaGny~pa~rGna~r~~r~r~~rl (SEQ.ID.NO.: d) Gl~sllee~r~~rIe~nh~ul~ge (SEQ.ID.NO.: 2), AMENDED, SHEET 1'9560Y PE-T/US 96/1649U 106 Rec'd PCTPTO -17 NOV 1997 -162- e) AsilLyslleSerTyrTyrISer (SEQ.ID. NO.: 127), f) AsnLysAdaSerTyrGlnlSer (SEQ.JD.NO.: 128), g) SerTyrGiniSerSer (SEQ.ID.NO.: 129), h) LysTyrGinISerSer (SEQ.ID.NO.: 140); i) hArgTyrGnSer~er (SEQ.ID.NO.: 141); j) hArgChaGlnlSer~er (SEQ.ID.NO.: 185); and k) TyrGiniSerSer (SEQ.ID.NO.: 186); wherein hArg is homoarginine and Xaa is any natural amino acid; XL is absent or is an amino acid selected from: a) leucine, b) isoleucine, c) norleucine, and d) valine; and R is acetyl, pivaloyl or benzoyl, or the pharmaceutically acceptable salt thereof.

9. The method of treatment according to Claim 7 wherein the conjugate is selected from: '19560Y 106Re'd PC T?'TC 17 NO0V 199? -163- OH 'CH 2 0H OH 3 NH x wherein X is: AsnLyslleSerTyrGlnSer- AsnLyslleSerTyrG InSerSer.. Asn LyslIleSerTyrGlInSerSerSer AsnLyslleSerTyrGlnSerSerSerThr AsnLyslleSerTyra !nSerSerSerThrGlu AlaAsnLysi IeSerTyrGlnSerSerSerTh rG Iu N-terminus (SEQ.ID.NO.: 13), (SEQ.ID.NO.: 16), (SEQ.ID.NO.: 17), (SEQ.ID.NO.: 3), (SEQ.ID.NO.: 11), AENDED ,S 19560Y Pcr/us 96/16490 'A V 799, -164- Ac AlaAsnLyslleSerTyrGlnSerSerSerThr- Ac AIaAsn Lys I IeSerTyrG InSerSerSerTh rLe- Ac -AlaAsnLysAaSer~yrGInSerAaSerThrLe* Ac AlaAsnLysAlaSerTyrGlnSerAlaSerLeu Ac AlaAsnLysAlaSerTyrG InSerSerSerLeu Ac AlaAsnLysAlaSerjyrG InSerSerLeu- Ac SerTyrGlnSerSerSerLeu Ac h~gyGnere~re (S Ac LysTyrGlnSerSerSerLeu (S Ac LysTyrGInSerSerNile (S N-terminus (SEQ.ID.NO.: 117), (SEQ.ID.NO.: (SEQ.ID.NO.: 118), (SEQ.ID.NO.: 119), (SEQ.ID.NO.: 120), (SEQ.ID.NO.: 121), ~EQ.ID.NO.: 144), EQ.ID.NO.: 145), ~EQ.ID.NO.: 124), or EQ.ID.NO.: 146), or the pharmaceutically acceptable salt thereof.

10. The method of treatment according to Claim 7 wherein the conjugate is selected from: Ac-hArgTyrGnSerSerPro-dox( 3 (SEQ.ID.NO.: 151) Ac-hArgTyrGlnSerPro-dox(3') (SEQ.ID.No.: 177) Ac-hArgTyrGln-SerSerSerNledo( 3 (SEQ.ID.NO.: 154) Ac-AmffyrGln-SerSerSerNle-dox( 3 (SEQ. ID.NO.: 155) H2C-~gyGnSre~re-o(' (SEQ.ID.No.: 156) AMENDED. SHEET -165- Ac-LysTyrGln-SerSerNle-.dox(3') (SEQ.JD.NO.: 146) Ac-LysTyrGln-.SerLysNe-.dox(3') (SEQ.JD.NO.: 178) Ac(cis-p-NH2Cha)TyrG~nSerSerNledox(3') (SEQ.LD.No.: 161) Ac-A~aAspLysAla(hg)TyrnSerSereu-dox( 3 (SEQ.ID.NO.: 160) Ac-hArgTyrGln-SerAsn.dox(3I) (SEQ.ID.NO.: 153) Ac-hArgTyrG~n..SerSer~s-dox(3I) (SEQ.ID.NO.: 152) Ac-(imidazolyl)LysTyrG~n-SerSerJeu-dox(3t) (SEQ.ID.NO.: 159) Ac-(imidazolyl)LysTyffGlnSerSerSerNle-dox(3') (SEQ.ID.NO.: 162) Ac-~r(Ch)Gn-Sr~r~e~l-do(3) (EQID.NO. 163) *10 Ac-hg(Me2PO3 Tyr)Gln-SerSerSerNle.dox(3') (SEQ. ID.NO.: 167) Ac-hArgTyrGln-SerserSerh~g-dox(3') (SEQ.ID 164) Ac-hArg(3 -Iodo-Tyr)Gln-SerS erSerNle-dox(3') (SEQ.ID.NO.: 166) Ac-hArg(O-Me-yr)Gln.SerSerSerNe-dox(3I) (SEQ. ID.NO.: 169) Ac-hArg(p-NI{2.Phe)Gln..SerserSerNle..dox(3y) (SEQ .ID.NO.: 179) Ac-hArg(Cha)Gln.SerSerNle.dox(3I) (SEQ.ID.NO.: 174) Ac-hArg(Cha)Gln.SerProNle-dox(3') (SEQ. ID. NO.: 175) Ac(imidazolyl)Lys(Cha)G~nSerSerSerNle-dox(3) (SEQ.ID.NO.: 172) Ach*g7 OTI)lnSe.r*rledx(' (SEQ.ID.NO.: 180) Ac-hArg( 3 -Fluoro)TyrGlnSerSerSerNle-do( 3 (SEQ.ID.No.: 176) Ac-(ornithine)TyrGln-.SerSerSerNledox(3) (SEQ.ID.NO.: 181) Ac-LysAlaAlaSerSerSerLeu-dox(3) (SEQ.ID.NO.: 182) Ac-hArgh(Cha)GlmS erSerNle-dox(3') (S EQ.ID 149) Ac-AlaArgLysAlaSerTyrGln-Seru-dox( 3 (SEQ.ID.NO.: 193) and Ac-(Om)TyrGin-SerSerS erLeu-dox(3') (SEQ.ID.NO.: 194) or the pharmaceutically acceptable salt thereof.

11. The method of treatment according to Claim 6 wherein the conjugate is of the formula 11: '19560Y pGTIUS 96116490 106 Rec'd PCTIPTO 17 NOV 1997 -166- CH 3 0- oligopeptide R wherein: oligopeptide is an oligopeptide which is specifically recognized by the free prostate specific antigen (PSA) and is capable of being proteolytically cleaved by the enzymatic activity of the free prostate specific antigen; XL is absent or is an amino acid selected from: a) phenylalanine, b) leucine, c) valine, d) isoleucine, e) 2 -naphthyl)alanine, f) cyclohexylalanine, g) diphenylalanine, h) norvaline, i) norleucine, and j) 1, 2 3 4 -tetrahydroisoquinoline3carboxylic acid; or XL is -NH -(CH2)n -NH AMEND~ED, SHUET 19560Y PCT/US 96/16490 106 Rec'd PCT/PTO 17 NO V 1997 -167- R is hydrogen or -(C=O)R1; R1 is C1-C6-alkyl or aryl; R 19 is hydrogen or acetyl; and n is 1, 2 3, 4 or or the pharmaceutically acceptable salt thereof. 0

12. The method of treatment according to Claim 6 wherein the conjugate is of the formula Ia: OH 'OCOCH 3 oligopeptide NH 2 C-terminus C-terminus wherein: oligopeptide is an oligopeptide which is specifically recognized by the free prostate specific antigen (PSA) and is capable of being proteolytically cleaved by the enzymatic activity of the free prostate specific antigen, AMENDED SHEET 168 or the pharmaceutically acceptable salt thereof.

13. The method of treatment according to claim 11 wherein the conjugate is: OH CH 3 0 e* S 5 S S. S 555 SS S 5555 S S S 55 55 S S 5*5* N terminus Ac- LysTyrGnSerSerSerNeNH (SEQ. ID. NO.: 183),

14. The method of treatment according to claim 12 wherein the conjugate is: *S55 S S S ,2' [R:\LIBAAJ0729 Idoc:TAB -169- OH N E t N CO 2 CH 3 H N '"'"CH2CH3 CH3. N OH S- CH 3 O H :CH 3 .O 5^ 0 ee HN- LeuAsnLysAlaSerTyrGnSerSerSerLeu-C-NH 2 Compound 5 (SEQ.ID.NO.: 184), or the pharmaceutically acceptable salt thereof. A method of treating an adverse condition of the 5 prostate which comprises administering to a mammal in need of said treatment a conjugate, said conjugate which comprises two pharmaceutical agents, wherein at least one pharmaceutical agent is effective against said condition, attached to a oligopeptide, wherein the oligopeptide comprises a sequence of amino acids that is recognized and selectively proteolytically cleaved by free prostate specific antigen, wherein the means of attachment is directly through a covalent bond or via a linker unit, or the pharmaceutically acceptable salt thereof.

16. A method of treating benign prostatic hyperplasia which comprises administering to a mammal in need of said treatment a conjugate, said conjugate which comprises two cytotoxic agents attached to an oligopeptide, wherein the oligopeptide comprises a sequence of -170- amino acids that is recognized and selectively proteolytically cleaved by free prostate specific antigen, wherein the means of attachment is directly through a covalent bond or via a linker unit, or the pharmaceutically acceptable salt thereof.

17. The method of treatment according to Claim 16 wherein the conjugate is a a 9* a a.. *aa a a. H CH 3 O CH 2 CH 3 "OH "i3 C=O C-terminus LeuAsnLysAlaSerTyrGInSerSerLeu OH (SEQ.ID.NO.: 184), I CH 3 Compound or the pharmaceutically acceptable salt thereof. L C^ -171-

18. A pharmaceutical composition useful for treating an adverse condition of the prostate comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a conjugate, said conjugate which comprises a pharmaceutical agent, effective in the treatment of said condition, attached to a oligopeptide, wherein the oligopeptide comprises a sequence of amino acids that is recognized and selectively proteolytically cleaved by free prostate specific antigen, wherein the means of attachment is directly through a covalent bond or via a linker unit, or the pharmaceutically acceptable salt thereof. 1 9. A pharmaceutical composition useful for treating benign prostatic hyperplasia comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a conjugate, said conjugate which comprises a cytotoxic agent attached to a oligopeptide, wherein the oligopeptide comprises a sequence of amino acids that is recognized and selectively proteolytically cleaved by free prostate specific antigen, wherein the means of attachment is directly through a covalent bond or via a linker unit, o S or the pharmaceutically acceptable salt thereof. The composition according to Claim 19 wherein the conjugate is of the formula I: y1 XC :.A 172 o OH 0 CH 2 0H OH CH 3 0 0 OH 0 OH 3 0 TONH OH XL Oligopeptide R wherein: oligopeptide is an oligopeptide which is specifically recognized by a a. the free prostate specific antigen (PSA) and is capable of being proteolytically cleaved by the enzymatic activity of the free prostate *aa. specific antigen; 10 XL is absent or is an amino acid selected from: a a) phenylalanine, b) leucine, c) valine, d) isoleucine, e) 2 -naphthyl)alanine, f) cyclohexylalanine, g) diphenylalanine, h) norvaline, i) norleucine, and j) 1,2,3 4 -tetrahydroisoquinoline-3-carboxylic acid; R is hydrogen or 1; and 0 90~000 0 0* 0 S .0 S. a 0 0 **0S 09 S 000 *0 0e *0 *050 OS Sq -b0 0 0 0S0~ 0000 0 1* *500 S *@SS0~ 173 R I is C1I-C6-alkyl or aryl, or the pharmaceutically acceptable salt thereof.

21. The composition according to Claim 19 wherein the conjugate is selected from: O OH 0 CH 2 0H 11 bH CH 3 0 0 0H60 OH 3 0 NH OH X wherein X is: AsnLysIIeSerTyrG3InSer AsnLyslleSerTyrGnSerSer... AsnLyslleSerTyrGlnSerSerSer AsnLyslleSerTyrGlnSerSerSerThr AsnLysIIeSerTyrGInSerSerSerThr~lu AlaAsnLyslIIeSerTyrGInSerSerSerThr.~lu- N-terminus (SEQ.ID.NO.: 13), (SEQ.ID.NO.: 16), (SEQ.ID.NO.: 17), (SEQ.ID.NO.:1O), (SEQ.ID.NO.: 3), (SEQ.JD.NO.: 11),

174- Ac ~AlaAsnLysleSerTyrGInSerSerSerThr-. Ac -AIaAsn Lys IIeSe ryryInSe rSerSerTh e+ Ac -AlaAsnLysAlaSerTyrGlniSerAlaSerThrLe&... Ac -AlaAsnLysAlaSerTyrGInSerAlaSereu... Ac AlaAsnLysAlaSerTyrG InSerSerSerLeu Ac AIaAsnLysAIaSerTyrGnSerSerLeu- Ac SerTyrGlnSerSerSerLeu Ac hArgTyrGlnSerSerSerLeu.... (SEQ.ID.NO.: 117), (SEQ.ID.NO.: (SEQ.ID.NO.: 118), (SEQ.ID.NO.: 119), (SEQ.ID.NO.: 120), (SEQ.ID.NO.: 121), 0 0 0e 0 0* 00 0 00 os S 0 0 S 00.. S 0300 *5 S @0S S. 0@ 00 0065 0e *0 0* 0 0 0 a OS.. OS'S 5 0 0000 0 005. 0 5 3EQ.ID.NO.: 144), 3EQ.ID.NO.: 145), Ac LysTyrGnSerSerSerLeu (SEQ.ID.NO.: 124), or Ac- LysTyrGinSerSerNte (SEQ.ID.NO.: 146), t N-terminus or the pharmaceutically acceptable salt thereof. 2 2. The composition according to Claim 19 wherein the conjugate is selected from: Ac-hArgTyrGnSerSerPro-dox(3') (SEQ.ID 151) Ac-hrgTyrGn-SerPro-.dox(3') (SEQ.ID.NO.: 177) Ac-hArgTyrGnSerSerS erNle-dox(3') (SEQ.ID 154) Ac-AmffyrGln-SerSerSerNle-dox(3 t) (SEQ.ID.NO.: 155) H2NCO-hsgTyrGln-SerSerSerLeu-dox(3') (SEQ.ID.No.: 156) Ac-LysTyrGn.SerSerNle-dox(31) (SEQ .ID.NO.: 146) 175 Ac-LysTyrGln-SerLysNle-.dox(3') (SEQ.JD.NO.: 178) Ac (cis-p-NH2Cha)TyrGlnSerSerNledox(3 t) (SEQ.ID.NO.: 161) AcAas~s~~~gTr~-e~re-o(' (SEQ.ID.NO.: 160) Ac-hArgTyrG~n-SerAsn..dox(3') (SEQ.ID.NO.: 153) Ac-hArgTyrG~nSerSeris-dox(3') (SEQ.ID.NO.: 152) Ac-(imidazoly1)LysTyrG~n.SerSerjeu-dox(3.) (SEQ.ID. NO.: 159) erSer~le-dox(3 t) (SEQ.ID.NO.: 162) Ac-hArg(Cha)Gln-serSerserNle.dox(3') (SEQ. ID 163) Ac-hArg(Me2PO3 Tyr)Gln-SerSerSerNle-dox(3') (SEQ.ID 167) Ac-hArgTyrG~n-SerSerSerh.Jg-dox(3') (SEQ.JD.NO.: 164) Ac-hArg(3 -Iodo-Tyr)Gln-SerSerSerNle-dox(3') (SEQ.ID 166) Ac-hArg(O-Me-.Tyr)Gln.S erSerSerNle-dox(3y) (SEQ. ID.NO.: 169) Ac-hArg(p-NH2-Phe)GlnSerSerSerNle-dox(3') (SEQ.ID 179) Ac-hArg(Cha)Gln-SerSerNe-.dox(3 t) (SEQ.ID.NO.: 174) Ac-hArg(Cha)Gln-SerProNle-dox(3') (SEQ.ID.NO.: 175) fl. Ac(imidazolyl)Lys(Cha)GlnSerS erSerNle-dox(3') (SEQ .ID.NO.: 172) Achr(-OTCGnSre~rl-o(' (SEQ.ID.NO.: 180) Ac-hArg( 3 -Fluoro)TyrG~nserSerSerNle-dox(3I) (SEQ.ID.NO.: 176) Ac-(ornithine)TyrCGln-.SerSerSerNledox(3y) (SEQ. ID NO.: 181) 20 Ac-LysAlaAlaSerSerSer~eu-.dox(3') (SEQ.ID.NO.I

182.) Ac-hArgh(Cha)Gln-SerSerNle-dox(3 t) (SEQ.ID 149) Ac-AlaArgLysAlaSerTyrG~n-Seru-.dox(3y) (SEQ.ID.NO.: 193) and Ac- (Orn)TyrGln-SerSerSerLeu-.dox(3') (SEQ.ID. NO.: 194) or the pharmaceutically acceptable salt thereof. 2 3. The composition according to Claim 19 wherein the conjugate is of the formula II: ,19560Y PUT/US 9A/16490 -176- 106 Rec'd PCTIPTO -7 NOV 1W OH N "H N HO CH 3 E II 0 XL -oligopeptide R wherein: oligopeptide is an oligopeptide which is specifically recognized by the free prostate specific antigen (PSA) and is capable of being proteolytically cleaved by the enzymatic activity of the free prostate specific antigen; XL is absent or is an amino acid selected from: a) phenylalanine, b) leucine, c) valine, d) isoleucine, e) 2 -naphthyl)alanine, f) cyclohexylalanine, g) diphenylalanine, h) norvaline, i) norleucine, and j) l, 2 3 4 -tetrahydroisoquinoline-3-.carboxylic acid; or XL is NH -(CH2)n -NH AMENDED, SH 177 R is hydrogen or 1; R I is C I-C6-alkyl or aryl; R 19 is hydrogen or acetyl; and n is 1, 2, 3, 4 or or the pharmaceutically acceptable salt thereof. conugte s:24. The composition according to Claim 2 3 wherein the 0 0 a. 0 I,,Et CH 3 0 N-terminus Ac- LysTyrGinSerSerSerfie NH or the pharmaceutically acceptable salt thereof. Compound 14 (SEQJID.NO.: 183), 2 5. The composition according to Claim 19 wherein the conjugate is of the formula 11: I 'I r -178- OH OCOCH 3 CH 3 CO oligopeptide NH 2 C-terinus C-terminus wherein: 5 oligopeptide is an oligopeptide which is specifically recognized by the free prostate specific antigen (PSA) and is capable of being proteolytically cleaved by the enzymatic activity of the free prostate specific antigen, or the pharmaceutically acceptable salt thereof. 26. The composition according to Claim 19 wherein the conjugate is: ',jhJ 3 :i _i -179- OH N Et CH 3 N-teri0Cnus H .N O LeuAsnLysAlaSerTyrGInSerSerSerLeu NH 2 Compound 5 (SEQ.ID.NO.: 184), or the pharmaceutically acceptable salt thereof. S27. A pharmaceutical composition useful for treating an 5 adverse condition of the prostate which comprises a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a conjugate, said conjugate which comprises two pharmaceutical agents, o wherein at least one pharmaceutical agent is effective against said condition, attached to an oligopeptide, wherein the oligopeptide comprises a sequence of amino acids that is recognized and selectively proteolytically cleaved by free prostate specific antigen, wherein the means of attachment is directly through a covalent bond or via a linker unit, or the pharmaceutically acceptable salt thereof. 28. A pharmaceutical composition useful for treating benign prostatic hyperplasia comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a conjugate, said 3 u -180- conjugate which comprises two cytotoxic agents attached to an oligopeptide, wherein the oligopeptide comprises a sequence of amino acids that is recognized and selectively proteolytically cleaved by free prostate specific antigen, wherein the means of attachment is a covalent bond or a chemical linker, or the pharmaceutically acceptable salt thereof. 29. The composition according to Claim 28 wherein the conjugate is a a. S S. S a a a. *5*a S a S a. a a CH 3 0 a a '3 0-=0 C-terminus LeuAsnLysAaSerryrGlnSer SerLeu (SEQ.ID.NO.: 184), OH \N OH 3 0 CH 3 0 0 OH 0 Compound 10 1 Ai I I OH 6 or the pharmaceutically acceptable salt thereof. I, 181 A pharmaceutical composition useful for treating an adverse condition of the prostate, substantially as hereinbefore described with reference to any one of the Examples. 31. Use of a conjugate which comprises a pharmaceutical agent, attached to an oligopeptide, wherein the oligopeptide comprises a sequence of amino acids that is recognised and selectively proteolyptically cleaved by free prostate specific antigen and wherein the means of attachment is directly through a covalent bond or via a linker unit, or the pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating an adverse condition of the prostate. 32. Use of a conjugate which comprises a pharmaceutical agent, attached to an oligopeptide, wherein the oligopeptide comprises a sequence of amino acids that is recognised and selectively proteolyptically cleaved by free prostate specific antigen and wherein the means of attachment is directly through a covalent bond or via a linker unit, or the pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating benign prostatic hyperplasia. 15 33. A medicament for treating an adverse condition of the prostate when prepared by the use of claim 31. 34. A medicament for treating benign prostatic hyperplasia when prepared by the use of claim 32. Dated 8 June, 1999 20 Merck Co., Inc. Patent Attorneys for the Applicant/Nominated Person SPRUSON FERGUSON B ~;r [R:\LIBAA]07291.doc:TAB