CA2234763A1 - Conjugates useful in the treatment of benign prostatic hyperplasia - Google Patents

Abstract

Novel pharmaceutical compositions 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 are described. Methods of treating benign prostate hypertrophy are also disclosed.

Description

CA 02234763 1998-04-1~

TITL~ OF THE INVENTION
CONJUGATES USEFUL IN THE TREATMENT O~ BENIGN
PROSTATIC HYPERPLASIA

5 BACKGROUN~ 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) 10 remain without symptoms, others show slow progression, while others remain stable. However, some 400,000 men a year have symptoms severe enough to require ~ur~e~y. The most common surgery, transurethral resection, is effective in relieving the symptoms of BPH, although side-effects, including morbidity from the operation itself, mild 15 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 45, when the tissue begins to change, growin2 and causin2 the size of the prostate to increase. The enlarging prostate squeezes the urethra, 20 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 25 in~ection 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 serninal fluid (Nadji, M., Taber, S.Z., Castro, A., et al. (1981) Cancer 30 48:1229;Papsidero, L., Kuriyama, M., Wang, M., et al. (1981). JNCI
66:37; Qui, S.D., Young, C.Y.F., Bihartz, D.L., et al. (1990), J. Urol.
144:1550; Wang, M.C., Valenzuela, L.A., Murphy, G.P., 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 .

W O 97/14416 PCT~US96/16490 molecular mass. PSA is a protease with chymotrypsin-like specificity (Christensson, A., Laurell, C.B., 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 sperrn entrapping gel, Semeno~elin I and Semenogelin II, and fibronectin (Lilja, H. (1985). J. Clin. Invest.
76: 1899; Lilja, H., Oldbring, J., Rannevik, G . . et al . (1987). J. Clin.
Invest. 80:281; McGee, R.S., Herr, J.C. (1988). Biol. Reprod. 39:499).
The PSA mediated proteolysis of the gel-formin~ proteins generates 10 several soluble Semenogelin I and Semenogelin II fragments and soluble fibronectin fragments with liquefaction of the ejaculate and release of progressively motile spermatoza (Lilja, H., Laurell. C.B. (1984). Scand.
J. Clin. Lab. Invest. 44:447; ~hcGee, R.S., Herr, J.C. (1987). Biol.
Reprod. 37:431). Furthermore, PSA may proteolytically degrade IGFE3P-15 3 (insulin-like growth factor binding protein 3) allowing IGF to stimulate specifically the glrowth 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 20 95~o of the detected serum PSA (Christensson. A., Bjork, T., Nilsson, O., et al. (1993). J. Urol. 1~0:100-105; Lilja, H., Christensson, A., Dahlén, U. (1991). Clin. Chem. 37:1618-1625; Stenman, U.H., Leinoven, J., Alfthan, H., et al. (1991). Cancer Res. 51 :222-226) . The prostatic tissue (normal, beni~;n hyperplastic, or malignant tissue) is implicated to 25 predomin~nt1y release the mature, enzymatically active form of PSA, as this form is required for complex formation with alpha l -antichymotrypsin (Mast, A.E., F.nghild, J.J., Pizzo, S.V., et al. (1991).
Biochemistry 30:1723-1730; Perlmutter, D.H., Glover, G.I., Rivetna, M., et al. (1990). Proc. Natl. Acad. Sci. USA 87:3753-3757). Therefore, in 30 the microenvironrnent of prostatic PSA secreting cells, the PSA is believed to be processed and secreted in its mature enzymatically active fonn not complexed to any inhibitory molecule. PSA also forrns stable complexes with alpha 2 - macroglobulin, but as this results in encapsulation of PSA and complete loss of the PSA epitopes, the in vivo -CA 02234763 1998-04-1~

significance of this complex formation is unclear. A free, noncomplexed form of PSA constitutes a minor fraction of the serum PSA
(Christensson, A., Bjork, T., Nilsson, O., et al. (1993). J. Urol. 150:100-105; Lilja, H., Christensson, A., Dahlén, U. (1991). Clin. Chem.
37:1618-1625). The size of this form of serum PSA is ~simil~r to that of PSA in seminal fluid (Lilja, H., Christensson, A., Dahlén, 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 lcDa forrn of PSA (Christensson, A., Bjork, T., Nilsson, O., et al. (1993). J.
IJrol. 150:100-105; Lilja, H., Christensson, A., Dahlén, 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). AbovenormalserumconcentrationsofPSAhave also been reported in benign prostatic hyperplasia and subsequent to surgical trauma of the prostate (Lilja, H., Christensson, A., Dahlén, 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 30 novel ph~ c eutical 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.

CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 Another object of this invention is to provide a method of treating benign prostatic hyperplasia which comprises ~-lmini~tration of the novel ph~ ceutical composition. ,.
S 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 10 pharmaceutical agent are described. Methods of treating such conditions of the prostate are also disclosed.
BRIEF DESCRIPTION OF THE FIGURES

15 FIGURES 1 and lA: 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 ("CS") are shown (numbered in order of the relative affinity of a site towards PSA
hydrolysis) and the protein fragments are numbered sequentially starting 20 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 25 various times. The results are shown in Table 2. All of the oligopeptides were tested as trifluoroacetate salts.

FIGURES 3, 3A and 3B: Cleavage A~inity of Synthetic Oligopeptides:
Synthetic oligopeptides were prepared and the oligopeptides were 30 digested with enzymatically active free PSA for four (4) 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 35 oligopeptide, the free base was tested.

CA 02234763 1998-04-1~

S

FIGURE 4: Cytotoxicity Data of Non-cleavable Oligopeptide-Doxorubicin Conjugates:
The data of the figure shows comparative cytotoxicity of doxorubicin and S 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 0.3, LM, while the acetylated oligopeptide modified doxorubicin has an Ec50 that has been reduced by greater than 300 fold. This conjugate had no HPLC detectable 10 cont~min~tion with unmodified doxorubicin. The oligopeptide alone had no detectable cell killing activity.

FIGURES S and 5A: Cleavage Affinity of Oligopeptides in Conjugation with Doxorubicin by Free PSA In Vitro:
lS Qligopeptides-doxorubicin conjugates were prepared and the conjugates were digested with enzymatically active free PSA for four (4) hours. The percentage conjugate that is enzymatically cleaved in the oligopeptide in this period of time is listed. The results are shown in Table S. Table Sa shows the amount of time (in minutes) required for 50% cleavage of the 20 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 25 Doxorubicin in Cell Conditioned Media:
Oligopeptides-doxorubicin conjugates were reacted for four (4) 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 30 cleaved in the oligopeptide in this period of time is listed. The results are shown in Table 6.

FIGUPE 7: Cytotoxicity Data of Cleavable Oligopeptide-Doxorubicin Conjugates:

CA 02234763 1998-04-1~
W O 97/14416 PCT~US96/16490 The data in Table 7 shows cytotoxicity (as EC50) 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 o~ 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.

I~)ETAILED DESCRIPTION OF THE INVENTION
The present invention relates to phannaceutical 10 compositions that comprise conjugates that contain oligopeptides, which are specifically recognized by the free prostate specific antigen (PSA) and are capable of being proteolyhcally cleaved by the enzymatic activity of the free prostate speci~lc antigen, and pharm~ceutical agents covalently ked to such oligopeptides directly or through a linker unit, or 15 pharmaceutically acceptable salts thereof. In particular, this invention is directed to such conjugates wherein the pharmaceutical a~ent 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 olicomers that comprise an amino acid sequence selected from:
a) AsnLysIleSerTyrGlnlSer (SEQ.ID.NO.: 13), 25 b) LysIleSerTyrGlnlSer (SEQ.ID.NO.: 14), c) GlyGluAsnGlyValGlnLysAspValSerGlnXaaSerIleTyrlSerGlnThrGlu (SEQ.ID.NO.: 15), 30 d) GlyLysGlyIleSerSerGlnTyrlSerAsnThrGluGluArgLeu (SEQ.ID.NO.: 2), e) AsnLysIleSerTyrTyrlSer (SEQ.ID.NO.: 127), CA 02234763 1998-04-1~

f) AsnLysAlaSerTyrGlnlSer (SEQ.ID.NO.: 128), g) SerTyrGlnlSerSer (SEQ.ID.NO.: 129);
5 h) LysTyrGlnlSerSer (SEQ.ID.NO.: 140);
i) hArgTyrGlnlSerSer (SEQ.ID.NO.: 141);

j) hArgChaGlnlSerSer (SEQ.ID.NO.: 185); and k) TyrGlnlSerSer (SEQ.ID.NO.: 186);

wherein hArg is homoarginine, Cha is cyclohexyl~l~nine and Xaa is any natural amino acid.

In an embodiment of the instant invention, the oligopeptides include oligomers that comprise an arnino acid sequence that is selected from:
20 a) AsnLysIleSerTyrGlnlSerSer (SEQ.ID.NO.: 16), b) AsnLysIleSerTyrGlnlSerAla (SEQ.ID.NO.: 130), c) AsnLysIleSerTyrGlnlSerSerSer (SEQ.ID.NO.: 17), d) AlaAsnLysIleSerTyrGlnlSerSerSer (SEQ.ID.NO.: 18), e) LysIleSerTyrGlnlSerSerSerThrGlu (SEQ.ID.NO.: 19), 30 f) GlyGluAsnGlyValGlnLysAspValSerGlnArgSerIleTyrlSerGlnThrGlu (SEQ.ID.NO.: 4), g) GlyGluAsnGlyValGlnLysAspValSerGlnSerSerIleTyrlSerGlnThrGlu (SEQ.ID.NO.: 5), CA 02234763 1998-04-1~

h) AlaAsnLysIleSerTyrTyrlSer (SEQ.ID.NO.: 131), i) AlaAsnLysAlaSerTyrGlnlSer (SEQ.ID.NO.: 132), s j) SerTyrGlnlSerSerThr (SEQ.ID.NO.: 133), k) SerTyrGlnlSerSerSer (SEQ.ID.NO.: 134), 10 1) LysTyrGlnlSerSerSer (SEQ.ID.NO.: 142), m) hArgTyrGlnlSerSerSer (SEQ.ID.NO.: 143), and n) SerTyrGlnlSerSerLeu (SEQ.ID.NO.: 135);
1~
or the phArm~ceutically acceptable salt thereof.

In a more preferred embodiment of the instant invention, the oligopeptides include oligomers that comprise an amino acid sequence 20 that is selected from:
a) AsnLysIleSerTyrGln ISerSerSerThr (SEQ.ID.NO.: 10), b) AlaAsnLysIleSerTyrGlnlSerAla (SEQ.ID.NO.: 136), c) AsnLysIleSerTyrGlnlSerSerSerThrGlu (SEQ.ID.NO.:3 ), d) AlaAsnLysIleSerTyrGlnlSerSerSerThrGlu (SEQ.ID.NO.: 11), 30 e) GlyGluAsnGlyValGlnLysAspValSerGlnArgSerIleTyrlSerGlnThrGlu (SEQ.ID.NO.: 4), f) AlaAsnLysIleSerTyrTyrlSerSer (SEQ.ID.NO.: 137), CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 -- 9 _ g) AlaAsnLysIleSerTyrTyrlSerAla (SEQ.ID.NO.: 138), h) AlaAsnLysAlaSerTyrGlnlSerAla (SEQ.ID.NO.: 139), 5 i) AlaSerTyrGlnlSerSerLeu (SEQ.ID.NO.: 94);
or the pharmaceutically acceptable salt thereof.

In a further embodiment of the instant invention, the 10 oligopeptides include oligomers that comprise an amino acid sequence that is selected from:

a) GlyArgLysAlaAsnLysIleSerTyrGlnlSerSerSerThrGluGluArgArg LeuHisTyr GlyGluAsnGly (SEQ.ID.NO.: 6).

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 20 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:
25 AsnLysIleSerTyrGlnlSerSerSerThr (SEQ.ID.NO.:10) 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 30 acid, or both terminus amino acids are modified. Such modifications include, but are not limited to, acylation of the amine group at the N-terminll~ and formation of an amide to replace the carboxylic acid at the C-terminn~. Addition of such moieties may be performed during solid-phase synthesis of the oligomer; thus, attachment of the C-terminll~

._ amino acid to a solid phase resin may be through an amine which results in an amide moiety upon acidic cleavage of the oligomer from ~e resin.
Thus the ~ollowing compounds are considered "oligomers that comprise an amino acid sequence" as used hereinabove and are meant to be 5 illustrative and are not limiting:

AlaAsnLysIleSerTyrGlnlSerSerSerThrGlu-amide (SEQ.ID.NO.: 11) Ac-AlaAsnLysIleSerTyrGlnlSerSerSerThrLeu (SEQ.ID.NO.: 70) lQ Ac-AlaAsnLysIleSerTyrGlnlSerSerSerThrGlu-amide (SEQ.ID.NO.: 11) Ac-AlaAsnLysIleSerTyrGlnlSerSerSerThrLeu-amide (SEQ.ID.NO.: 70) Ac-AlaAsnLysIleSerTyrGlnlSerAlaSerThrGlu-amide (SEQ.ID.NO.: 73) Ac-AlaAsnLysIleSerTyrGlnlSerSerLysThrGlu-amide (SEQ.ID.NO.: 74) Ac-AlaAsnLysIleSerTyrGlnlSerSerThrGlu-amide (SEQ.ID.NO.: 75) 15 Ac-AlaAsnLysIleSerTyrGlnlSerSerGlnThrGlu-amide (SEQ.ID.NO.: 78) Ac-AlaAsnLysIleSerTyrGlnlSerAlaLysThrGlu-amide (SEQ.ID.NO.:79) Ac-AlaAsnLysIleSerTyrGlnlSerThrGlu-arnide (SEQ.ID.NO.: 81) Ac-AlaAsnLysSerTyrGlnlSerSerThrGlu-amide (SEQ.ID.NO.: 82) Ac-AlaAsnLysAlaSerTyrGlnlSerAlaSerThrGlu-~mide (SEQ.ID.NO.:
20 84) Ac-AlaAsnGluIleSerTyrGlnlSerAlaSerThrGlu-amide (SEQ.ID.NO.: 85) Ac-AsnEysIleSerTyrGlnlSerSer-amide (SEQ.ID.NO.: 16) Ac-LysIleSerTyrGlnlSerSer-arnide (SEQ.ID.NO.: 86) Ac-SerTyrGlnlSerSerThrGlu-amide (SEQ.ID.NO.: 87) 25 Ac-AlaSerTyrGlnlSerSerThrGlu-amide (SEQ.ID.NO.: 89) Ac-AlaAsnLysIleSerTyrTyrlSerSerSerThrGlu-amide (SEQ.ID.NO.: 92) Ac-AlaAsnLysIleSerTyrTyrlSerAlaSerThrGlu-amide (SEQ.ID.NO.: 93) Ac-AlaSerTyrGlnlSerSerLeu-amide (SEQ.ID.NO.: 94) Ac-AlaAsnSerTyrGlnlSerSerSerThrGlu-amide ~SEQ.ID.NO.: 95) 30 Ac-AlaSerTyrGlnlSerSerSerThrGlu-amide (SEQ.ID.NO.: 96) Ac-SerTyrGlnlSerSerSerThrGlu-amide (SEQ.ID.NO.: 97) Ac-AlaAsnLysAlaSerTyrGlnlSerAlaSerCys-amide (SEQ.ID.NO.: 98) Ac-hArg(Cha)GlnlSerNle-Acid (SEQ.ID.NO.: 147) Ac-hArghTyrGlnlSerSerNle-Acid (SEQ.ID.NO.: 148) CA 02234763 1998-04-1~

Ac-hArgh(Cha)GlnlSerSerNle-Acid (SEQ.ID.N0.: 149) Ac-AlaAspLysAlaSerTyrGlnlSerSer-Cha-NHNH2 (SEQ.ID.N0.: 150) Ac-hArgTyrGlnlSerSerPro-Acid (SEQ.ID.N0.: 151) Ac-hArgTyrGlnlSerSerHis-Acid (SEQ.ID.N0.:152 ) 5 Ac-hArgTyrGlnlSerAsn-Acid (SEQ.ID.N0.: 153) Ac-hArgTyrGlnlSerSerSerNle-Acid (SEQ.ID.N0.: 154) Ac-(Amf)TyrGlnlSerSerSerNle-Acid (SEQ.ID.N0.: 155) H2NCO-hArgTyrGlnlSerSerSerLeu-Acid (SEQ.ID.N0.: 156) Ac-AlaAspLysAlaLysTyrGlnlSerSer(Cha)-NHNH2 (SEQ.ID.N0.: 157) 10 Ac-(DPL)TyrGlnlSerSerSerNle-Acid (SEQ.ID.N0.: 158) Ac-(imi~1~701e)LysTyrGlnlSerSerLeu-Acid (SEQ.ID.N0.: 159) Ac-AlaAspLysAla(hArg)TyrGlnlSerSerLeu-Acid (SEQ.ID.N0.: 160) Ac-(p-NH2-Cha)TyrGlnlSerSerSerNle-Acid (SEQ.ID .N0.: 161) Ac(imidazolyl)LysTyrGlnlSerSerSerNle-Acid (SEQ.ID.N0.: 162) 15 Ac-hArg(Cha)GlnlSerSerSerNle-Acid (SEQ.ID.N0.: 163) Ac-hArgTyrGlnlSerSerSerhArg-Acid (SEQ.ID.N0.: 164) Ac-hArgTyrGlnlSerSerSer(MeLeu) (SEQ.ID.N0.: 188) Ac-hArgTyrGlnlSerSerSer(Ethylester-Leu) (SEQ.ID.N0.: 156) Ac-AlaAspLysAla(imidazoleLys)TyrGlnlSerSerNle-Acid (SEQ.ID.N0.:
20 165) Ac-hArg(3-Iodo-Tyr)GlnlSerSerSerNle-Acid (SEQ.ID.N0.: 166) Ac-hArg(Me2P03-Tyr)GlnlSerSerSerNle-Acid (SEQ.ID.N0.: 167) Ac-hArgTyrGlnlSerSerAsp-Acid (SEQ.ID.N0.: 168) Ac-hArg(0-Me-Tyr)GlnlSerSerSerNle-Acid (SEQ.ID.N0.: 169) 25 Ac-AlaAspLysAlaLysTyrGlnlSerSerNle-Acid (SEQ.ID.N0.: 170) Ac-hArg(Cha)GlnlSerSerSer(ethylester-Leu) (SEQ.ID.N0.: 171) Ac-(imidazolyl)Lys(Cha)GlnlSerSerSerNle-Acid (SEQ.ID.N0.: 172) Ac-hArg(Cha)GlnlSerSerSer-Acid (SEQ.ID.N0.: 173) Ac-hArg(Cha)GlnlSerSerNle-Acid (SEQ.ID.N0.: 174) 30 Ac-hArg(Cha)GlnlSerProNle-Acid (SEQ.ID.N0.: 175) and Ac-hArg(m-fluoro-Tyr)GlnlSerSerSerNle-Acid (SEQ.ID.N0.: 176), .

or the ph~nT ~ceutically acceptable salt thereof.

CA 02234763 1998-04-1~

A person of ordinary skill in the peptide chemistry art would readily appreciate that certain arnino 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 5 oligopeptide has been conserved in the modified oligopeptide. Certain llnn~tllral 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 3-iodotyrosine, 2-methyltyrosine, 3-fluorotyrosine, 3-methyltyrosine and 10 the like. Further for example, lysine may be replaced with N'-(2-imidazolyl)lysine and the like. The following list of amino acid replacements is meant to be illustrative and is not limiting:

Ori~inal Amino Acid Replacement Amino Acid(s) Ala Gly Arg Lys, Ornithine Asn Gln Asp Glu Glu Asp Gln Asn Gly Ala Ile Val, Leu, Met, Nle Leu Ile, Val, Met, Nle Lys Arg, Ornithine Met Leu, Ile, Nle, Val Ornithine Lys, Arg Phe Tyr, Trp Ser Thr Thr Ser Trp Phe, Tyr Tyr Phe, Trp Val Leu, Ile, Met, Nle CA 02234763 1998-04-1~

W O 97/14416 PCTnUS96/16490 ~ 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:

5 AsnArgIleSerTyrGlnlSer (SEQ.ID.NO.: 21) AsnLysValSerTyrGlnlSer (SEQ.ID.NO.: 22) AsnLysMetSerTyrGlnlSerSer (SEQ.ID.NO.: 23) AsnLysLeuSerTyrGln ISerSer (SEQ.ID.NO.: 24) AsnLysIleThrTyrGlnlSerSerSer (SEQ.ID.NO.: 25) 10 AsnLysIleSerPheGlnlSerSerSer (SEQ.ID.NO.: 26) AsnLysIleSerTrpGlnlSerSerSerThr (SEQ.ID.NO.: 27) AsnLysIleSerTyrAsnlSerSerSerThr (SEQ.ID.NO.: 28) AsnLysIleSerTyrGlnlThrSerSerThr (SEQ.ID.NO.: 29) AsnLysIleSerTyrGlnlSer (SEQ.ID.NO.: 30) 15 ~lnLysIleSerTyrGlnlSerSer (SEQ.ID.NO.: 31 ) AsnArgIleThrTyrGlnlSerSerSer (SEQ.ID.NO. 32) AsnArgIleSerPheGlnlSerSerSerThr (SEQ.ID.NO.: 33) AsnArgIleSerTrpGlnlSerSerSerThr (SEQ.ID.NO.: 35) AsnArgIleSerTyrGlnlThrSerSerThr (SEQ.ID.NO.: 36) 20 AsnLysIleThrTyrGlnlThrSerSerThr (SEQ.ID.NO.: 37) AsnLysLeuSerTyrGlnlThrSerSerThr (SEQ.ID.NO.: 38) GlnLysLeuSerTyrGlnlSerSerSerThr (SEQ.ID.NO.: 39) AsnArgLeuSerTyrGlnlThrSerSerThr (SEQ.ID.NO.: 40) AsnLysValSerPheGlnlSerSerSerThr (SEQ.ID.NO.: 41) 25 AsnArgValSerTrpGlnlSerSerSerThr (SEQ.ID.NO.: 42) GlnLysValSerTyrGlnlSerSerSerThr (SEQ.ID.NO.: 43) GlnLysIleSerTyrGlnlThrSerSerThr (SEQ.ID.NO.: 34) AsnLysIleSerTyrGlnlSerSerSerThr (SEQ.ID.NO.: 44);

30 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:

CA 02234763 1998-04-l~

W O 97/14416 PCT~US96/16490 GlyGluGlnGlyValGlnLysAspValSerGlnSerSerIleTyrlSerGlnThrGlu (SEQ.ID.NO.: 45), GlyGluAsnGlyLeuGlnLysAspValSerGlnSerSerIleTyrlSerGlnThrGlu 5 (SEQ.ID.NO.: 47), GlyGluAsnGlyValAsnLysAspValSerGlnSerSerIleTyrlSerGlnThrGlu (SEQ.ID.NO.: 48), GlyGluAsnGlyValGlnArgAspValSerGlnArgSerI leTyrlSerGlnThrGlu (SEQ.ID.NO.: 49), 10 GlyGluAsnGlyValGlnLysAspValSerGlnL,ysSerIleTvrlSerGlnThrGlu (SEQ.ID.NO.: 50), GlyGluAsnGlyValGlnLysAspLeuSerGlnThrSerIleTyrlSerGlnThrGlu (SEQ.ID.NO.: 51 ), GlyGluAsnGlyValGlnLysAspValSerGlnSerSerIlePhelSerGlnThrGlu 15 ~SEQ.ID.NO.: 52), GlyGluAsnGlyValGlnLysAspMetSerGlnSerSerIleTyrlThrGlnThrGlu (SEQ.ID.NO.: 53), GlyGluAsnGlyValGlnLysAspValSerGlnAr~SerIleTyrlThrGlnThrGlu (SEQ.ID.NO.: 54), 20 GlyGluAsnGlyValGlnLysAspValSerGlnSerSerIleTyrlSerGlnSerGlu (SEQ.ID.NO.: 55), GlyGluAsnGlyValGlnLysAspValSerGlnArgSerIleTyrlSerAsnThrGlu (SEQ.ID.NO.: 56), GlyLysAlaIleSerSerGlnTyrlSerAsnThrGluGluArgLeu (SEQ.ID.NO.:
25 57), GlyArgGlyIleSerSerGlnTyrlSerAsnThrGluGluArgLeu (SEQ.ID.NO.:
59), GlyLysGlyIleThrSerGlnTyrlSerAsnThrGluGluAr~Leu (SEQ.ID.NO.:
60), 30 GlyLysGlyIleSerThrGlnTyrlSerAsnThrGluGluArgLeu (SEQ.ID.NO.:
61), GlyLysGlyIleSerSerAsnTyrlSerAsnThrGluGluAr~Leu (SEQ.ID.NO.:
62), -CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 -- AlaLysGlyIleSerSerGlnTyrlSerAsnThrGluGluArgLeu (SEQ.ID.NO.: 63), GlyLysGlyIleSerSerGlnPhelSerAsnThrGluGluArgLeu (SEQ.ID.NO.:
64), 5 GlyLysGlyIleSerSerGlnTyrlThrAsnThrGluGluAr~Leu (SEQ.ID.NO.:
65), GlyLysGlyIleSerSerGlnTyrlSerAsnSerGluGluArgLeu (SEQ.ID.NO.:
58), and GlyLysGlyIleSerSerGlnTyrlSerAsnThrAspGluArgLeu (SEQ.ID.NO.:
10 46);
and the like.
The inclusion of the symbol "I" within an amino acid sequence indicates the point within that sequence where the oligopeptide is proteolytically cleaved by free PSA.
15 ~ 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"
20 stereoconfi2uration The following abbreviations are utilized in the specification and figures to denote the indicated amino acids and moieties:

hR or hArg: homoarginine hY orhTyr: homotyrosine Cha: cyclohexyl~ nine Amf: 4-aminomethylphenyl~l~nine DPL: 2-(4,6-dimethylpyrimidinyl)lysine (imidazolyl)K: N'-(2-irnidazolyl)lysine Me2PO3-Y: O-dimethylphosphotyrosine O-Me-Y: O-methyltyrosine TIC: tetrahydro-3-isoquinoline carboxylic acid MeL: 2-keto-3-amino-5-methylhexane DAP: 1,3-tli~minopropane CA 02234763 1998-04-1~

TFA: trifluoroacetic acid AA: acetic acid The method of treatment of the instant invention utilizes 5 pharm~ce~tical compositions whose ph~ eutical 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 10 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 15 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 20 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 cont~ining the PSA proteolytic cleavage site is bonded directly, or through a chemical linker, to the cytotoxic 25 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 30 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 , CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 physiological 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 5 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 10 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.
Bec~ e the conjugates utilized in the instant invention can be used for modifying a given biological response, cytotoxic agent is not 15 t~ be construed as limite~l to classical chemical therapeutic agents. For example, the cytotoxic agent may be a protein or polypeptide possessing a desired biological activity. Such proteins rnay include, for example, a toxin such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin; a protein such as tumor necrosis factor, oc-interferon, ,B-interferon, nerve 20 growth factor, platelet derived growth factor, tissue plasminogen activator; or, biological response modifiers such as, for example, lymphokines, interleukin-l ("IL-l"), interleukin-2 ("IL-2"j, interleukin-6 ("IL-6"), granulocyte macrophage colony stimulating factor ("GM-CSF"), granulocyte colony stim~ ting factor ("G-CSF"), or other growth 25 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 30 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, c~rminomycin, daunorubicin, aminopterin, methotrexate, methopterin, dichloro-methotrexate, mitomycin C, porfiromycin, 5-fluorouracil, 6-mercaptopurine, cytosine arabinoside, podophyllotoxin, or podo-phyllotoxin derivatives such as etoposide or etoposide phosphate, melphalan, vinblastine, vincristine, leurosidine, vindesine, leurosine, taxol and the like. Other useful cytotoxic agents include estramustine, S cisplatin and cyclophosph~mi~le. 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 hi~hly preferred group of cytotoxic a~ents for the present 10 invention include drugs of the following formulae:

THE METHOTREXATE GROUP OF FORMULA( 1):

H2N ~,N~,N~ R7 >=~ COR9 ~ N~ ~ CONHCHCH2C H2CO2H

(1) in which R12 is amino or hydroxy;
R7 is hydrogen or methyl;
R8 is hydrogen, fluoro, chloro, bromo or iodo;
R9 is hydroxy or a moiety which completes a salt of the carboxylic acid;

W O 97/14416 PCT~US96/16490 - THE MITOMYCIN GROUP OF FORMULA (2):
O
H2N J~ CH20CONH2 H3C~N2 o N' R10

(2) in which R 10 is hydrogen or methyl;
s THE BLEOMYCIN GROUP OF FORMULA (3) Ix H~CONH2 ~
N N HO CH3 H ~ N
H2N~O ~ )~N~NH N~

CH3 H~N CH3 )\ ~S

~ ~1 ~----o1 ~ N
HO~
OH ¦~OH
HO o OH

(3) 10 in which Rl 1 is hydroxy, amino, Cl-C3 aLkylamino, di(Cl-C3 aLkyl)amino, C4-C6 polymethylene amino, NH
+ 11 -NHCH2CH2CH2S-CH3; or-NHCH2CH2CH2CH2NH-C-NH2;

MELPHALAN OF FORMULA (4):

NH2 N(CH2CH2c1)2

(4) 6-MERCAPTOPURINE OF FORMULA (5):

N~'~NH>

(5) A CYTOSINE ARABINOSIDE OF FORMULA (6):

N O
HOH2C ~

HO OH

(6) -T~F PODOPHYLLOTOXINS OF FORMULA(7):

R13 ~o ~
0~o\
HO ~~
<o~O

( ) CH30J~oCH3 OH
S i~ which R13 is hydrogen or methyl;
R14 is methyl or thienyl;
or a phosphate salt thereof;

THE~ VINCA ALKALOID GROUP OF DRUGS OF FORMULA (8): -N~ ." R17 ~COzCH3 ~ J~,CH2CH3 CH30 F.~ OR 19 co2CH3 - (8) s in which R15 is H, CH3 or CHO; when R l 7 and R l 8 are taken singly;
R18 is H, and one of R16 and R17 is ethyl and the other is H
or OH; when R17 and R18 are taken together with the carbons to which they are attached, they form an oxirane ring in which case R l 6 is ethyl;
Rl9 is hydrogen, (Cl-C3 aLkyl)-CO, or chlorosubstituted (Cl-C3 aLkyl)-CO;

15 nIFLUORONUCLEOSIDES OF FORMULA (9):

R2l~,0~CH20H

F OH

(9) in which R21 is a base of one of the form W O 97/14416 PCT~US96/16490 -- O O R

O~N~ H2N ~¢N~ O N

N~ N~N~

in which R22 is hydrogen, methyl, bromo, fluoro, chloro or iodo;
R23 is -OH or-NH2;
R24 is hydrogen, bromo, chloro or iodo;
or, THE ANTHRACYCLINES ANTIBIOTICS OF FORMULA (10):

R3 O OH o CH3~
R6'R~ R4 (1 o) .
wherem R1 is -CH3, -CH20H, -CH20CO(CH2)3CH3, or -CH20COCH(OC2H5)2;

R3 is -OCH3, -OH or-H;
R4 is -NH~, -NHCOCF3, 4-morpholinyl, 3-cyano-4-morpholinyl, l-piperidinyl, 4-methoxy-1-piperidinyl, benzyl~mine, dibenzylamine, cyanomethylamine, or 1-cyano-2-methoxyethyl amine;
R5 is -OH -OTHP or-H; and R6 is -OH or -H provided that R6 is not -OH when RS is -OH or -OTHP.

10 l~STRAMUSTINE (1 ] ) CH OH

(CICH2CH2)2NCOO ~J
(1 1 ) CYCLOPHOSPHAMIDE (12) o~ , N(cH2cH2cl)2 I O
NH

The most highly preferred drugs are the anthracycline antiobiotic agents of Formula (10), described previously. One skilled in 20 the art understands that this structural formula includes compounds which are drugs, or are derivatives of drugs, which have acquired in the art ~lirrer~lt 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.

W O 97/14416 PCT~US96/16490 1 ~ 5 ~ ~ ~ O

~1 0 0 0 0 0 0 ~ ~ O

o X X ~ ~ ~: X ~ ~
~Z Z Z Z Z Z Z Z Z
_ o=~ ~~ ~
~ I ~ D a v ~, ~ c~ c~ c~ c~ ~
o~o ~~10 0 0 0 ~ ~ ~ ~ ~ .G

O O O O O O C~
~I C~ ~ C~ C~ C~ C~ C~

" . _ U~
. ~ D ~~ ~- G

E~ ~ e, ~ e ~ ", x ~ ~ ~

CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 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 Forrnula (10) in which Rl is -cH2oH7 R3 is 5 -OCH3, R4 is -NH2~ R5 is -OH, and R6 is -H.

The oligopeptides, peptide subunits and peptide derivatives (also terrned "peptides") incorporated in the conjugates utilized in the method of treatment of the present invention c~n be synthesized from 10 their constituent amino acids by conventional peptide synthesis techniques, preferably by solid-phase technolog~ . The peptides are then purified by reverse-phase high performance liquid chromatography (HPLC).
Standard methods of peptide synthesis are disclosed, for 15 example, in the following works: Schroeder et al., "The Peptides", Vol. I, Acadernic Press 1965; Bodansky et al., "Peptide Synthesis", Interscience Publishers, 1966; McOmie (ed.) "Protective Groups in Organic Chemistry", Plenum Press, 1973; Barany et al., "The Peptides: Analysis, Synthesis, Biology" 2, Chapter 1, Academic Press, 1980, and Stewart ef 20 al., "Solid Phase Peptide Synthesis", Second Edition, Pierce Chemical Company, 1984. The teachings of these works are hereby incorporated by reference.
The ph~r~n~ceutically acceptable salts of the compounds incorporated in the conjugates lltili~l in the method of treatment of 25 this invention include the conventional non-toxic salts of the compounds of this invention as formed, e.g., from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and 30 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, slllf~nilic, 2-acetoxy-benzoic, fumaric, CA 02234763 1998-04-1~

toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic and the like.
The conjugates lltili7e-1 in the method of treatment of the instant invention which comprise the oligopeptide cont~ining the PSA
5 cleavage site and a cytotoxic agent may .5imil~rly 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 forrned. Similarly, an amide bond may be forrned by covalently coupling 10 an amine moiety of the oligopeptide and a carboxyl moiety of the cytotoxic agent. For these purposes a reagent such as 2-(lH-benzotriazol- l-yl)- 1 ,3,3-tetramethyluronium hexafluorophosphate (known as HBTU) and l-hyroxybenzotriazole hydrate (known as HOBT), dicyclohexyl- carbodiimide (DCC), N-ethyl-N-(3-15 d~methylaminopropyl)- carbodiimide (EDC), diphenylphosphorylazide (DPPA), benzotriazol- 1 -yl-oxy-tris-(dimethylamino)phosphonium hexafluorophosphate (BOP) and the like, used in combination or singularly, may be lltili7e-1 Furthermore, the instant conjugate may be forrned by a non-20 peptidyl bond between the PSA cleavage site and a cytotoxic agent. Forexample, 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 25 imicl~7ole, a combination of DCC and DMAP, and the like may be lltili7e~1 The carboxylic acid may also be activated by forming the nitro-phenyl ester or the like and reacted in the presence of DBU (1,8-diazabicyclo[5 ,4,0]undec-7-ene.
The instant conjugate may also be formed by attachment of 30 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 forrning an amide bond. Conversely, CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 a tli~minoalkyl diradical linker unit, whereby a carbonyl moiety on the cyctotoxic agent is covalently attacted to one of the amines of the linker unit whi]e 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 10 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 synthesi~ of conjugates utilized in the method of treatment of the invention, one may need to protect or block various reactive functionalities on the starting 15 compounds and int~rmediates 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 exarllple, hydrolytic or hydrogenolytic means. Such protection and deprotection steps are conventional in organic chemistry. One 20 skilled in the art is referred to Protective Groups in Or~anic Chemistry.
McOmie, ed., Plenum Press, NY, NY (1973); and, Protective Groups in Or~anic 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 arnino-protecting groups may include, for example, Cl-Clo aLkanoyl groups such as formyl, acetyl, dichloroacetyl, propionyl, hexanoyl, 3,3-diethylhexanoyl, ~-chlorobutryl, and the like; Cl-Clo alkoxycarbonyl and C5-Cls aryloxycarbonyl groups such as tert-butoxycarbonyl, benzyloxycarbonyl, 30 allyloxycarbonyl, 4-nitrobenzyloxycarbonyl, fluorenylmethyloxycarbonyl and cinnamoyloxycarbonyl; halo-(Cl-Clo)-aLkoxycarbonyl such as 2,2,2-trichloroethoxycarbonyl; and Cl-C15 arylalkyl and aLkenyl ~roup such as benzyl, phenethyl, allyl, trityl, and the like. Other comrnonly -CA 02234763 1998-04-1~

W O 97/14416 PCTrUS96/16490 used amino-protecting groups are those in the form of en~min~s prepared with ,B-keto-esters such as methyl or ethyl acetozl~et~te.
Useful carboxy-protecting groups may include, for example, Cl-Clo aLkyl groups such as methyl, tert-butyl, decyl; halo-cl-clo alkyl S such as 2,2,2-trichloroethyl, and 2-iodoethyl; C5-C1s arylalkyl such as benzyl, 4-methoxybenzyl, 4-nitrobenzyl, triphenylmethyl, diphenyl-methyl; Cl-Clo alkanoyloxymethyl such as acetoxymethyl, propionoxymethyl and the like; and groups such as phenacyl, 4-halophenacyl, allyl, dimethylallyl, tri-(Cl-C3 aL~yl)silyl, such as 10 trimethylsilyl, ~-p-toluenesulfonylethyl, ,B-p-nitrophenyl-thioethyl, 2,4,6-trimethylbenzyl, ~-methylthioethyl, phth~limidomethyl. 2,4-dinitro-phenylsulphenyl, 2-nitrobenzhydryl and related groups.
Similarly, useful hydroxy protecting groups may include, ~or example, the formyl group, the chloroacetyl group, the benzyl group, the 15 benzhydryl 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 20 anthracycline antibiotic doxorubicin, the following Reaction Schemes illustrate the synthsis of the conjugates of the instant invention.

PCT~US96/16490 REACTION SCHEME I

~ z ~ H20H

CH30 ~ OH o dox CH30 ~ OH O
CH3- ~ 12 ~ H--oligopeptide ¦
OH
C-terrninus = H20 1 oligopeptide ¦

CH30 ~ OH ~ C-terrninus CH3- ~"~g 13 OH

W O 97/14416 PCT~US96/16490 REACTION SCHEME ll C::: [ ~ ~H20H

CH30 ~ OH o CH3~ H20H

11 OH CH30 ~ OH o CH3- ~ NH~-protect C-terminus O OHHN I oligopeptide ¦
~ ~H20H

CH30 ~ OH o CH3- ~g 14 ~' NH2 OH

W O 97/14416 PCT~US96/16490 REACTION SCHEME lll ~ H70H

CH30 ~ \ Oq~ (CH2)2C02H

CH30 0 OH o 1 1 "~

/ CH ~ (~/

OH
H O

~, ~N~I oligopeptide ¦
~J ~,J ~OH CH20H
CH30 ~ OH o CH3~ 1 5 OH

REACTION SCHEME IV

~ ~ ~ H20H

CH30 0 OH o ~' NH2 OH

H2NHN I oligopeptide ¦
C-terminus C-terrninus H 1~

O OH N--N I oligopeptide CH30 ~ OH o CH3 ~~g 16 OH

W O 97/14416 PCTrUS96/16490 REACTION SCHEME V

~ ~H3 Br CH30 0 OH o CH3~

OH
/ HS~

t C-terminus CH~ ; H I ~li9opeptide ¦

ol C-terminus CH
f' NH2 OH

-CA 02234763 1998-04-1~

W O 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 5 illustrated (S.P. ~n~ ri et al. J. Med. Chem. 28:1079-lOg8 (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 10 use of the 1,3-~ minopropane 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-~-position hydroxy moiety is reacetylated following the addition of the linker unit. Applicants have 15 discovered that the ~les~cetyl vinblastine conjugate is also efficacious and may be prepared by elimin~ting 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 20 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 25 of treatment is combined with one cytotoxic agent, such as vinblastine, while the C-terrninus 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 30 advantages over a conjugate cont~inin~ only one cytotoxic agent.

REACTION SCHEME VI

OH
~Et N2H4, reflux, MeOH

,N02CH3 H ~

I ' ""' CH2CH3 CH30 ~~ N~ OCOCH3 co2CH3 vinblastine rN~ HONO

~ ~1 CH2CH3 r CH30 N~--OH

_~,. N~
T "' CH2CH3 CH30 ~N~--OH
- OH

W O 97/14416 PCT~US96/16490 REACTION SCHEME VI (Continued) OH
N~Et C-terminus H 1. oligopeptide- R
~NJ~Jco2cH3 2. Ac20, pyridine ,~' CH2CH3 CH30 N~OH

~3' CH2CH3 CH30 N~--OCOCH3 CO oligopeptide - NH2 C-terminus wherein R is -NH2, -O-alkyl and the like W O 9~/14416 PCTAUS96/16490 REACTION SCHEME VII

OH
N~\ ~ Et g~ 'H 2. BOC-CI
H

CH30~c~\~OH
3 _ ~ '~3 I ""CH2CH3 1. Ac20, pyridine CH30~N~OH
CH3 2. aq. HCI

O~ N NH - BOC

r ~
~'1 CH2CH3 CH30--~N, ~OCOCH3

7 - 39 -- REACTION SCHEME VII (Contin R'- oligopeptide C-terminus OH

H
,~,CH2CH3 CH30~--N \~OH OCOCH3 N~ N--oligopeptide - R' H
C-terminus wherein R' is acetyl, alkyl, hydrogen or the like S

W O 97/14416 PCT~US96/16490 REACTION SCHEME VIII

OHi_t 1. oligopeptide rN~ ~ 2. Ac20, pyridine OzCH3 or C-terminus r 1 ~ 1 oiigopeptide - OCH3 ~--~'""CH2CH3 2. LiOH
CH30~CH~ ~~ 3. Ac20, pyridine r ~
CH2CH3 doxorubicin CH30 ,N \~OCOCH3 CO - oligopeptide C-terminus W O 97/14416 PCTnUS96/16490 REACTION SCHEME VIII (Continued) OH

~2CH3 ~ ~' CH2CH3 CH30 N~OCOCH3 \
- OH /~~ opeptide CH3- ~ N-terminus CH30 0 OH ~

O~H

W O 97/14416 PCT~US96/16490 The oligopeptide-cytotoxic agent conjugate utili7ed 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:
s O OH O
~H20H

CH30 0 OH o ,~
CH3~1 ~H

OH XL - oligopeptide- R

wherem:
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) phenyl~l~nine, b) leucine, c) valine, d) isoleucine, e) (2-naphthyl)~ ninç, f) cyclohexyl~l~nine, g) diphenyl~ nine, h) norvaline, i) norleucine, and CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 j) 1,2,3,4-tetrahydroiso quinoline-3-carboxylic acid;
R is hydrogen or -(C=O)R1; and S Rl is Cl-c6-aL~yl or aryl, or the ph~ ce~ltically acceptable salt thereof.

In a preferred embodiment of the instant method of 10 treatment of BPH:

oligopeptide is an oligomer that comprises an amino acid sequence selected from:
15 a) AsnLysIleSerTyrGlnlSer (SEQ.ID.NO.: 13), b) LysIleSerTyrGlnlSer (SEQ.ID.NO.: 14), c) GlyGluAsnGlyValGlnLysAspValSerGlnXaaSerIleTyrlSerGlnThrGlu 20 (SEQ.ID.NO.: 15), d) GlyLysGlyIleSerSerGlnTyrlSerAsnThrGluGluArgLeu (SEQ.ID.NO.: 2), 2~ e) AsnLysIleSerTyrTyrlSer (SEQ.ID.NO.: 127), f~ AsnLysAlaSerTyrGlnlSer (SEQ.ID.NO.: 128), g) SerTyrGlnlSerSer (SEQ.ID.NO.: 129), h) LysTyrGlnlSerSer (SEQ.ID.NO.: 140);
i) hArgTyrGlnlSerSer (SEQ.ID.NO.: 141);

j) hArgChaGlnlSerSer (SEQ.ID.NO.: 185); and k) TyrGlnlSerSer (SEQ.ID.NO.: 186);

5 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, 1~ or the ph~rm~ceutically acceptable salt thereof.

The following compounds are specific examples of the oligopeptide-cytotoxic agent conjugate utilized in the method of 20 treatment of the instant invention:

W O 97/14416 PCT~US96/16490 " ~J ~ ,H20H

CH30 ~ OH o CH ~ ~
~/NH
OH X

wherein X is:

AsnLyslleSerTyrGlnSer-- (SEQ.ID.NO.: 13), AsnLyslleSerTyrGlnSerSer--(SEQ.ID.NO.: 16), AsnLyslleSerTyrGlnSerSerSer--(SEQ.ID.NO.:17 ), AsnLyslleSerTyrGlnSerSerSerThr--(SEQ.ID.NO.:10), AsnLyslleSerTyrGlnSerSerSerThrGlu-- (SEQ.ID.NO.: 3), AlaAsnLyslleSerTyrGlnSerSerSerThrGlu-- (SEQ.ID.NO.: 1 1 ), N-terminus W O 97/14416 PCT~US96/16490 Ac- AlaAsnLyslleSerTyrGlnSerSerSerThr - (SEQ.ID.NO.:117), Ac AlaAsnLyslleSerTyrGlnSerSerSerThrLeu - (SEQ.ID.NO.:70), Ac AlaAsnLysAlaSerTyrGlnSerAlaSerThrLeu - (SEQ.ID.NO.:118), Ac AlaAsnLysAlaSerTyrGlnSerAlaSerLeu - (SEQ.ID.NO.:ll9), Ac AlaAsnLysAlaSerTyrGlnSerSerSerLeu - (SEQ.ID.NO.:120), Ac AlaAsnLysAlaSerTyrGlnSerSerLeu - (SEQ.ID.NO.:121).

Ac SerTyrGlnSerSerSerLeu - (SEQ.ID.NO.:144), Ac hArgTyrGlnSerSerSerLeu - (SEQ.ID.NO.:145).

Ac LysTyrGlnSerSerSerLeu - (SEQ.ID.NO.:124), or (Compound 4) Ac - LysTyrGlnSerSerNle - (SEQ.ID.NO.:146).
N-terminus or the ph~ ceutical1y 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'-arnine are as follows: -10 Ac-hArgTyrGln-SerSerPro-dox(3') (SEQ.ID.NO.: 151) Ac-hArgTyrGln-SerPro-dox(3') (SEQ.ID.NO.: 177) Ac-hArgTyrGln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 154) CA 02234763 1998-04-l~

Ac-AmfTyrGln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 155) H2NCO-hArgTyrGln-SerSerSerLeu-dox(3') (SEQ.ID.NO.: 156) Ac-LysTyrGln-SerSerNle-dox(3') (SEQ.ID.NO.: 146) Ac-LysTyrGln-SerLysNle-dox(3') (SEQ.ID.NO.: 178) Ac(cis-p-NH2Cha)TyrGlnSerSerNledox(3') (SEQ.ID.NO.: 161) Ac-AlaAspLysAla(hArg)TyrGln-SerSerLeu-dox(3') (SEQ.ID.NO.: 160) Ac-hArgTyrGln-SerAsn-dox(3') (SEQ.ID.NO.: 153) Ac-hArgTyrGln-SerSerHis-dox(3') (SEQ.ID.NO.: 152) Ac-(imidazolyl)LysTyrGln-SerSerLeu-dox(3') (SEQ.ID.NO.: 159) 10 Ac-(imidazolyl)LysTyrGlnSerSerSerNle-dox(3 ') (SEQ.ID.NO.: 162) Ac-hArg(Cha)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 163) Ac-hArg(Me2PO3Tyr)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 167) Ac-hArgTyrGln-SerSerSerhArg-dox(3') (SEQ.ID.NO.: 164) Ac-hArg(3-Iodo-Tyr)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 166) 1~ Ac-hArg(O-Me-Tyr)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 169) Ac-hArg(p-NH2-Phe)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 179) Ac-hArg(Cha)Gln-SerSerNle-dox(3') (SEQ.ID.NO.: 174) Ac-hArg(Cha)Gln-SerProNle-dox(3') (SEQ.ID.NO.: 175) Ac(imidazolyl)Lys(Cha)GlnSerSerSerNle-dox(3') (SEQ.ID.NO.: 172) 20 Ac-hArg(7-HO-TIC)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 180) Ac-hArg(3-Fluoro)TyrGlnSerSerSerNle-dox(3') (SEQ.ID.NO.: 176) Ac-(ornithine)TyrGln-SerSerSerNle-dox(3 ') (SEQ.ID.NO.: 181) Ac-LysAlaAlaSerSerSerLeu-dox(3') (SEQ.ID.NO.: 183) Ac-hArgh(Cha)Gln-SerSerNle-dox(3') (SEQ.ID.NO.: 149) 2~ Ac-AlaArgLysAlaSerTyrGln-SerLeu-dox(3') (SEQ.ID.NO.: 193) and Ac-(Orn)TyrGln-SerSerSerLeu-dox(3') (SEQ.ID.NO.: 194) or the ph~rm~ceutically acceptable salt thereof.

The oligopeptide-cytotoxic agent conjugate ~ltili7ed in the method of treatment of the instant invention wherein the cytotoxic agent is the preferred cytotoxic agent vinblastine or desacetylvinblastine may be described by the general formula I below:

~ CA O 2 2 3 47 6 3 19 9 ~ Pc~uss6/l6490 ~Q 9'7/14416 Et ~ r~ 3 ,,~3,.,c~J2ct~3 C~3O CH. c)R19 ~0 X/ - Oligopeptide able of be~

Specific antigen;
b t or is an amino aCid se a) phenylalanine7 b) leucin c) valine.
d) isole~lcine~ .
e) (2 naphd~YI)alanln ' f~ CyclOhexylalanm g) diphenylalanine h) no~aline~ and i) nl 2 3 4-tetrahydroisoquino XL is ~ - (cH~)n R is hydrogen or -(C=O)Rl;

Rl is Cl-C6-aLkyl or aryl;

Rl9 is hydrogen or acetyl; and n isl,2,3,40rS, 10 or the ph~ eutically acceptable salt thereof.

The following compounds are specific examples of the oligopeptide-desacetylvinblastine conjugate utilized in the method of treatment of the in~t~nt invention:
lg OH
N~\ ~Et ~COzCH3 r ~
~/ ~" CH2CH3 CH30~N~OH

O~'NH
N-terminus ~ Compound 14 Ac--LysTyrGlnSerSerSerNle--NH (SEQ.ID.NO.: 183), o 2~3 47 63 199# pcTlus96/l649o W~ ~1144l6 3 ~
C~2C~3 C~ O~J-~~
Ct~3 C-term~nus ~o / L Sp~aSer~YrGlnSerSe (SEQ 1~ ~~- 184)-comP~~Ind eatment of ~he W O 97/14416 PCT~US96/16490 OH
N~\ .~ Et ~, CO2CH3 H , ~N~
j~' CH2CH3 CH30 N~OH
3 C =O C-terminus LeuAsnLysAlaSerTyrGlnSerSerLeu (SEQ.ID.NO.: 184), OH NH
CH3~

CH30 o OH ~
Compound 10 ~ l~fCH20H

~ OH O

or the ph~ ceutically acceptable salt thereof.

It is well known in the art, and understood in the instant invention, that peptidyl therapeutic agents such as the oligopeptide-cytotoxic agent conjugates preferably have the terminal amino moiety of any oligopeptide substituent protected with a suitable protecting group, lO such as acetyl, benzoyl, pivaloyl and the like. Such protection of the terminal amino group reduces or çlimin~t~s the enzymatic degradation of such peptidyl therapeutic agents by the action of exogenous amino CA 02234763 1998-04-l~

peptidases which are present in the blood plasma of warm blooded ~nim~1 e.
The oligopeptide-cytotoxic agent conjugates 1Itili7e~1 in the method of treatment of the instant invention are ~lmini~ctered to the 5 patient in the form of a ph~rrn~ceutical composition which comprises a conjugate of Formula (I) and a ph~rm~ceutically 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 ~nim~l including, for example, a human, equine, procine, 10 bovine, murine, canine, feline, or other m~mm~l, as well as an avian or other warrn-blooded ~nim~l. The preferred mode of ~-lminictration is parenterally, particularly by the intravenous, intramuscular, subcutaneous, intraperitoneal, or intralymphatic route. Such fo~n~ tions can be prepared using carriers, diluents or excipients f~mili~r to one 15- skilled in the art. In this regard, ~ee, ~g. Remin~ton's Pharmaceutical ~ciences, 16th ed., 1980, Mack Publishing Company, edited by Osol et al. Such compositions may include proteins, such as serum proteins, for example, h~ n serum albumin, buffers or buffering substances such as phosphates, other salts, or electrolytes, and the like. Suitable diluents 20 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 25 composition can include about 0.05 to about .20 percent by weight of an antioxidant such as sodium metabisulfite or sodium bisulfite.
For intravenous ~flminictration, the composition preferably will be prepared so that the amount ~f1minictered to the patient will be from about .01 to about 1 g of the conjugate. Preferably, the amount 30 ~ minietered 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 m~nner in which the conjugate is ~rlmini~etered~ the age, weight and condition of the patient as well as other factors to be determined by the treating physician. Thus, the amount ~rlmini~tered to any given patient must be determined on an individual basis.
One skilled in the art will appreciate that although specific 5 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.
FXAMPLES

EXAMPLE I

15- Identif~cation of the Semenogelin PSA Mediated Cleavage Site:
Liquefaction of the seminal gel parallels proteolytic fragmentation of semenogelin I [Lilja, H., Laurell, C.B., (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, H., (1985) J. Clin. Invest. 76, 1899-1903]. Uti1i7.in~ the published seguence of semenogelin I ~Lilja, H., Abr~h~mcson, P.A., Lundwall, A., (1989) J. of Biol. Chem. 264, 1894-1900] (Figure 1) we de.cigne-l polymerase chain reaction primers to clone the semenogelin cDNA from a commercially available prostatic cDNA library (Clone-tech, Palo Alto, CA.). The purified semenogelin cDNA was placed into the bacterial expression vector pTAC [Linemeyer, D.L., Kelly, L.J., Minke, J.G., Gimenez-Gallego, G., 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 30 semenogelin proteinThe bacterially expressed semenogelin protein was pllnf;e~l on an anti-tubulin antibody column. The purified semenogelin I
pl~tt~ 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 rnM Tris pH 8.0, 25 rnM NaCl, CA 02234763 1998-04-1~

- 54 ~
0.5 mM 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, P., (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 identi~led by variable time digestion, a l O minute digestion reaction was performed. The affinity of PSA for the S 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 st~inin~ intensity of each PSA generated peptide fragment. These intensities had approximate 15- ratios of 3: 1 :0.6:0.3.

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 colurnns using an aqueous 0.1 % trifluoroacetic acidlacetonitrile 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 ~l~ared by this method are shown in Figure 2.

CA 02234763 1998-04-1~

Assessment of the Recognition of Oligopeptides by Free PSA:
The oligopeptides prepared as described in Example 2 were individually S dissolved in PSA digestion buffer (1'~ mM tris(hydroxymethyl)-aminomethane pH8.0, 25 mM NaCl, 0.5 mM Cacl2) 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 NaCl. The reaction is quenched after various reaction 10 times by the addition of trifluoroacetic acid (TFA) to a ~mal 1%
(volume/volume). Alternatively the reaction is quenched with 10mM
ZnCl2. The quenched reaction was analyzed by HPLC on a reversed-phase C 18 column using an aqueous 0.1 %TFA/acetonitrile gradient. The results of the assessment are shown in Figure 2. Other oligopeptides 1~ ~re~aLed 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 signi~lcant loss of PSA mediated peptide hydrolysis, while the presence of a glutamic 20 acid residue at the carboxyl end of the peptide appears not to be essential to recognition by PSA.

25 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 synthesi~ or commercially available (Sigma)) in DMSO was added 30 HBTU and HOBT along with diisopropylethyl~mine and the reaction mixture was stirred overnight. The crude reaction mixture was pnrifie~l directly by preparative HPLC on a reversed-phase C-18 columrl using a 0.1% trifluoroacetic acid (TFA) in acetonitrile/0. 1% TFA in water gradient.

W O 97/14416 PCT~US96/16490 Table 3 ~ ~H20H

CH30 0 OH o CH3- ~
¦' NHR
OH

Compound R MS (parention) 12a H-Ala- 615 12b N-Ac-Ala- 657 12c N-Ac-Ala-Ala-Ala- 799.5 12d N-Ac-Ala-Gly-Pro-Thr-Gly-Ala-Ser- 1199 Ala-(SEQ.ID.NO.: 12) In vitro Assay of Cytotoxicity of Peptidyl Derivatives of Doxorubicin:
The cytotoxicities of ~e 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 15 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 cont~ininp: various concentrations of a given conjugate (final plate well volume of 200,u1). The cells were CA 02234763 1998-04-1~

W O 97/14416 PCTrUS96/16490 incubated for 3 days at 37~C and then 20~1 of Alamar Blue was added to the assay well. The cells were further incllb~te~ 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 5 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).

Assessment of Enzymaticnlly Active PSA from LNCaP Cells Enzymatic activity was demonstrated by incubating LNCaP serum free 15 media (concentrated approximately 200 fold) with recombinant Sememogelin I protein. Approximately 0.5 ,~Lg of immllnologically reactive PSA in concentrated conditioned media [deterrnined by HYBRIDTECH (Tandem E) elisa] was mixed with approximately 3 ,ug of recombinant Semenogelin I and incubated for 4 hours at 37~C. At the 20 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 25 produce detectable levels of circulating PSA.

Preparation of Cleavable Oligopeptide-Doxorubicin Conjugates:
30 The derivatives of doxorubicin wherein an oligopeptide which is proteolytically cleaved by free PSA is covalently attached to the arnine of the sugar moiety of the doxorubicin were prepared using the following general reaction: To a mixture of doxorubicin (Sigma) and ~e corresponding peptide (prepared by solid phase synthesis as described in Example 2) in DMSO was added HBTU and HOBT along with diisopropylethylamine and the reaction rnixture stirred overnight. The crude reaction mixture was puri~led directly by preparative HPLC on a reversed-phase C-18 colurnn using a 0.1 ~o trifluoroacetic acid (TFA) in S acetonitrile/0. 1% TFA in water gradient. When reactive arnine moieties were present on ~e peptide, such a functionality was typically protected as the fluorenylmethyloxycarbonyl adduct, which was removed by treatment with a secondary arnine, such as piperidine and the like, subsequent to conjugation with doxirubicin. The instant conjugates have 10 a structure of the general formula --~ ~ X i H20H

CH30 0 OH o NH
OH ¦ (O=C)peptide(NH) ¦
\

Ac and may be represented by the phrase "Ac-peptide-DOX (3')."
Conjugates which were prepared by the above general method or by the synthetic route described in Exarnple 8, but utili7ing the a~lo~liate 15 starting amino acid residues which are readily available cornrnercially or by synthetic techniques well known in the art, are listed in Tables 5, 5a and 7 in Figures 5, SA and 7.

Ac-I~ys-Tyr-Gln-Ser-Ser-Ser-Leu-Dox-Acetate CA 02234763 1998-04-l~

W O 97/14416 PCTrUS96/16490 _ 59 _ r Step A: Ac-Lys(Fmoc)-Gln-Ser(Bzl)-Ser(Bzl)-Ser(Bzl)-Leu-PAM
Resin (1).
Starting with 0.5 mmol (0.67g) Boc-Leu-PAM resin, the protected peptide was synthesized on a 430A ABI peptide synthesizer.
S The protocol used a 4 fold excess (2 m~nol) 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 (1).

Step B: Ac-Lys(Fmoc)-Tyr-Gln-Ser-Ser-Ser-Leu-OH (2) lS The protected peptide resin (1), 1.3 g, was treated with HF
(20 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 lliLuLated with H2O. The insoluble product (2) was filtered and dried (0.46g).

Step C: Ac-Lys(Fmoc)-Tyr-Gln-Ser-Ser-Ser-Leu-Dox (3) The above prepared intermediate (2), 0.46g, (0.43 mmol) was dissolved in DMF (15 ml) and doxorubicin hydrochloride, 125 mg (0.215 mrnol), added followed by 60 ,ul of triethyl~mine (0.430 mmol).
The stirred solution was cooled (0~C) and 92 ~11 of diphenylphosphoryl azide (0.43 mmol) added. After 5 minutes, an additional 92 ~11 of DPPA
was added and the pH adjusted to ~7.5 (pH paper) with TEA. After 1 hour, an additional 92 ,ul of DPPA was added, pH adjusted to ~7.5, 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 (3).
Step D: Ac-Lys-C~ln-Tyr-Ser-Ser-Ser-Leu-Dox (4).

W O 97/14416 PCT~US96/16490 The above product (3) was dissolved in DMF (20 ml), cooled (0~C) and 10 rnl of piperidine ~-lde~l The solution was concentrated to dryness and purified by preparative HPLC. Buffer A =
15% acetic acid-H20; B = 15% acetic acid-methanol. The crude product 5 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 l5~Lm, 300A). A step gradient of 10% B to 60% B was used at a flow rate of 75 mVmin (uv = 260 nm). Homogeneous product fractions were pooled, concentrated and freeze-dried from H2O to yield 125 mg of 10 purified product (4).

Deacetylvinblastinyl-Leu-Asn-Lys-Ala-Ser-Try-Gln-Ser-Ser-Ser-Leu-1~ NH2-Acetate (5) (SEQ.ID.NO. 184) Step A: NH2 - Leu-Asn-Lys(Fmoc)-Ala-Ser-Tyr-Gln-Ser-Ser-Ser-Leu-Arnide (6) Starting with 0.~ mmol of p-methylbenzhydrylamine resin 20 (MBHA), the protected peptide, NH2-Leu-Asn-Lys(Fmoc)-Ala-Ser(OBzl)-Tyr(BrZ)-Gln-Ser(OBzl)-Ser(OBzl)-Ser(OBzl)-Leu-MBHA, intermetli~tt 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-25 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 30 diisopropylethylarnine. 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 (7~ ml) was concentrated to dryness, dissolved in a 1:1 mixture of acetonitrile-H2O and freeze-dried to give 750 mg of crude product. A

CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 portion (200 mg) was pllrifi~l by preparative HPLC on a C-18 reverse phase support (Waters"u-Bondapak). Buffer A = 15% acetic acid-H20;
B = 15~o acetic acid-methanol. For the purification, the crude product was suspended in 400 ml of 10% B/90% A buffer, filtered and the filtrate S loaded onto the column. A step gradient of 10% B to 55~o B was used at a flow rate of 75 ml/min. Homogeneous product fractions were pooled, concentrated and freeze-dried from H2O to yield (6).

Step B: Deacetylvinblastin Monohvdrazide (7) lg of vinblastine sulfate was converted to the aII~ine form by extraction in methylene chloride and saturated sodium bicarbonate. The methylene chloride layer was washed with H2O, dried over anhydrous MgSO4 and concentrated to dryness. The vinblastine was then dissolved in anhydrous ethanol (20 ml) and anhydrous hydrazine added (20 rnl).
1~ 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 (7) was isolated. [Ref: K.S.P. Bhllsh~n~ Rao et al., J. Med.
Chem. (1985), 28:1079.]
Step C: Deacetylvinblastine Acid Azide (8).
Deacetylvinblastine monohydrazide (7) (48 mg, 0.0624 mmol) was dissolved in DMF (3 ml), cooled (-15~ C) and acidified to ~
2.5 (pH paper) with HCl/dioxane. Isoamylnitrite (10 ~11) was added 25 followed by an additional 10 ~Ll after 10 min. HPLC analysis indicated complete conversion of the hydrazide to azide after S min. The azide was m~int~in~.d in solution at -15~ C until ready for use.

Step D: Deacetylvinblastinyl-Leu-Asn-Lys-Ala-Ser-Try-Gln-Ser-Ser-Ser-Leu-NH~-Acetate (~) The oligopeptide product (6) from Step A, 32 mg (0.0225 rnmol), was dissolved in DMF (1 ml) and cooled (-15~ C). To this solution was added a 1.5 ml DMF solution (0.031 mmol) of desacetylvinblastine acid azide (8). The pH was adjusted to ~ 7.5 (pH

W O 97/14416 PCT~US96/16490 paper) wit~ triethylamine and the reaction stirred at -5~ C (2 hr), and 0~ C
for 18 hr. To the reaction was added H2O (2 rnl) and the solution evaporated to dryness. The intermediate was dissolved in DMF (4 ml), cooled (0~ 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 H2O to yield (5).

Deacetylvinblastinyl-Leu-Asn-Lys-Ala-Ser-Try-Gln-Ser-Ser-Ser-Leu--Dox-Acetate (10).
Step A: Deacetylvinblastinyl-Leu-Asn-Lys(Fmoc)-Ala-15- Ser-Try-Gln-Ser-Ser-Ser-Leu-Dox ~Acetate (9) The oligopeptide product (6) 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 (8) prepared as described in E~arnple 9, Step C. The pH was adjusted to ~ 7.5 (pH paper) with triethyl~mine and the reaction stirred at -15~ C for 90 mins.

After stirring 18 hours at 0-5~ C, the reaction was concentrated to dryness and the crude residue was dissolved in DMF (10 ml) and filtered. Doxorubicin hydrochloride, 62 mg (0.106 rnrnol), was added to the filtrate followed by 30 ,ul of triethylamine. The stirred solution was cooled (0~C) and 27 ,ul of diphenylphosphoryl azide (DPPA, 0.134 mmol) ~ l After S minutes, an additional 27 ,ul of DPPA was added and the pH adjusted to ~7.5 (pH paper) with TEA. After 1 hour, an additional 27 ~11 of DPPA was added, pH adjusted to -7.5, 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 (9).

Step B: Deacetylvinblastinyl-Leu-Asn-Lys-Ala-Ser-Try-Gln-Ser-Ser-Ser-Leu--Dox ~Acetate (10).

CA 02234763 1998-04-1~

The above intermediate product (9) was dissolved in DMF
(20 ml), cooled (0~C) and 10 ml of piperidine added. The solution was concentrated to dryness and purified by preparative HPLC. Buffer A =
15% acetic acid-H2O; 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, ~-Bondapak). A step gradient of 10% B to 60% B was used at a flow rate of 75 ml/min (uv = 260 nm). Semi-pure product was further purified on C-18 (Waters, Prep Pak) using Buffer A = 0.13M pH 3.0 10 triethylammonium phosphate and Buffer B = acetonitrile. A step 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% H2O (1% acetic lS acid). The product fractions were concentrated and freeze dried from H2O to yield the purified product (10).

~XAMPLE 1 1 20 Ac-Lys-Tyr-Gln-Ser-Ser-Ser-Nle-NH-(CH2)3 NH-deacetylvinblastine amide (14) Step A: Deacetylvinblastine-3-aminopropyl amide (11) To a cooled (-15~ C) a DMF solution (3 ml, 0.0624 mmol) of 25 deacetylvinblastine acid azide (synthesis described in Example 9, Step C) was added 1''0 ,ul of 1,3-~i~minopropane in DMF (2 rnl). The reaction was stirred at - 10~ C for 1 hr, filtered and concentrated to dryness to yield (11).

30 Step B: Deacetylvinblastine-3-aminopropylamide-norleucine amide (12) To a DMF solution (1 ml) of Boc-Nle (22 mg, 0.095 mmol) was added 318 ,ul of a lM solution of HOBT (in NMP) followed by 280 ,ul of a lM solution of DCC (in NMP). After 30 min., intennç~ te (11) (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: CH2cl2 (20 ml). After 5 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 ~c TFA-CH3CN. The crude product was loaded in 100% A buffer (100 ml) 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-Lys(Fmoc)-Tyr-Gln-Ser-Ser-Ser-Nle-OH ( 13) The above interrnediate was prepared as described in Example 9, Step Afor the preparation of Ac-Lys(Fmoc)-Tyr-Gln-Ser-Ser-lS Ser-Leu-OH.

Step D: Ac-Lys-Tyr-Gln-Ser-Ser-Ser-Nle-NH-(CH2)3 NH-deacetylvinblastine amide (14) The oligopeptide product (13), (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 (0~ C) and 17 ,~Ll of diphenylphosphoryl azide (0.08 mmol) added. After 5 min. an additional 17 ,ul of DPPA was added and the pH
adjusted to ~ 7.5 (pH paper) with triethylamine. After 2 hr. additional (13), 35 mg, was added in DMF (0.5 ml) and 17 ~LI of DPPA. The pH
was m~in~ined 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 (0~ 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. Purificahon ~, .
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).

Assessment of the Recognition of Oligopeptide-Doxorubicin Conjugates by Free PSA:
The conjugates prepared as described in Examples 7-9 were individually 10 dissolved in PSA digestion buffer (12 mM tris(hydroxymethyl)-aminomethane pH8.0, 25 mM NaCl, 0.5 mM CaCl2) and the solution added to PSA at a molar ration of 100 to 1. Alternatively, the PSA
digestion buffer lltili7e-1 is 50 mM tris(hydroxymethyl)-aminomethane pH7.4, 140 mM NaCl. The reaction is quenched after various reaction 15 times by the addition of trifluoroacetic acid (TFA) to a final 1%
(volume/volume). Alternatively the reaction is quenched with lOmM
ZnC12. The quenched reaction was analyzed by HPLC on a reversed-phase C18 column using an aqueous 0.1 %TFA/acetonitrile gradient. The results of the assessment are shown in Tables S and Sa of Figure 5.

Assessment of the Cleavage of Oligopeptide-Doxorubicin Conjugates in Cell Conditioned Media:
25 Cell conditioned serum-free a-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(~) Cent~ epTM concentrator with a 10,000 molecular weight cutoff. The 30 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.

CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 100 ~L portions of concentrated conditioned media was mixed with 35 ,~Lg of a oligopeptide-doxorubicin conjugate prepared as described in Example 7 and the mixture was incubated at 37~C for 0, 4 and 24 hour time points. The reactions were stopped by the addition of ZnC12 (to a 5 O.OlM final concentration) and analyzed by HPLC on a reversed-phase C 18 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.

In vitro Assay of Cytotoxicity of Peptidyl Derivatives of Doxorubicin:
The cytotoxicities of the cleaveable oligopeptide-doxorubicin conjl~g~tes, 15 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 20 conjugate (final plate well volume of 200~1). The cells were incubated for 3 days at 37~C, 20, L1 of Alamar Blue is added to the assay well. The cells were further incubated and the assay plates were read on a EL-3 10 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 25 various concentration of conjugate tested was then calculated versus control (no conjugate) cultures. Cytotoxicities of the conjugates were also compared to the cytotoxicity of umnodi~led do~orubicin and unmodified oligopeptide assessed in the same assay. Results of this assay are shown in Table 7 of Figure 7.

CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 F,~AMPLE 15 In vivo Ef~icacy of Peptidyl -Cytotoxic Agent Conjugates LNCaP.FGC or DuPRO- 1 cells are trypsinized, resuspended in the 5 growth medium and centifuged for 6 mins. at 200xg. The cells are resuspended in serum-free ~--MEM and counted. The a~r(j~liate volume of this solution cont~ining the desired number of cells is then transferred to a conical centrifuge tube, centrifuged as before and resuspended in the ~ypropliate volume of a cold 1:1 mixture of a--MEM-10 Matrigel. The suspension is kept on ice until the ~nim~l~ 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 5x105 DuPRO cells or 1.5x107 LNCaP.FGC cells.

Following inoculation with the tumor cells the mice are treated under one of two protocols:
Protocol A:
20 One day after cell inoculation the ~nim~l~ 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 maximllm non-lethal amount, but may be subsequently titrated lower. Identical doses are ~lnnini.ctered at 24 hour intervals for 5 days. After 10 days, blood 25 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 ~nim~l~' weights are determined at the beginning and end of the assay.
Protocol B:
Ten days after cell inoculation,blood samples are removed from the ~nim~l.c and serum levels of PSA are determined. ~nim~l.c are then grouped according to their PSA serum levels. At 14-15 days after cell W O 97/14416 PCT~US96/16490 inoculation, the ~nim~l~ 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 initi~lly the maximum non-lethal amount, but may be subsequently titrated lower. Identical doses are ~lmini~tered 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 ~nim~ ' weights are determined at the beginning and end of the assay.

CA 02234763 1998-04-1~

W O 97/14416 PCTnUS96/16490 SEQUENCE LISTING

(l) GENERAL INFORMATION:
(i) APPLICANT: DeFeo-Jones, Deborah Garsky, Victor M.
Jones, Raymond E.
Oliff, Allen I.
Scolnick, Edward M.
(ii) TITLE OF LNv~NllON: CONJUGATES USEFUL IN THE TREATMENT OF BENIGN
PROSTATIC HYPERPLASIA
(iii) NUMBER OF ~U~N~S: 194 (iv) CORRESPONDENCE ADDRESS:
(A) ADDRESSEE: DAVID A. MUTHARD
(B) STREET: 126 E. Lincoln Avenue, P.O BOX 2000 (C) CITY: RAHWAY
(D) STATE: NEW JERSEY
(E) COUNTRY: U.S.A.
(F) ZIP: 07065 (v) COMPUTER READABLE FORM:
(A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS
(D) SOFTWARE: PatentIn Release #l.0, Version #l.30 (vi) CURRENT APPLICATION DATA:
(A) APPLICATION NUMBER:
(B) FILING DATE:
(C) CLASSIFICATION:
(viii) AllOKN~Y/AGENT INFORMATION:
(A) NAME: Muthard, David A.
(B) REGISTRATION NUMBER: 35,297 (C) REFERENCE/DOCKET NUMBER: l9560 (ix) TELECOMMUNICATION INFORMATION:
(A) TELEPHONE: (908)594-3903 (B) TELEFAX: (908)594-4720 (2) INFORMATION FOR SEQ ID NO:l:
(i) ~Qu~N~ CHARACTERISTICS:
(A) LENGTH: 462 amino acids (B) TYPE: amino acid ~ (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO

CA 02234763 l998-04-l~

WO 97/14416 PCT~US96/16490 (v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:
Met Lys Pro Asn Ile Ile Phe Val Leu Ser Leu Leu Leu Ile Leu Glu ~ys Gln Ala Ala Val Met Gly Gln Lys Gly Gly Ser Lys Gly Arg Leu Pro Ser Glu Phe Ser Gln Phe Pro His Gly Gln Lys Gly Gln His Tyr Ser Gly Gln Lys Gly Lys Gln Gln Thr Glu Ser Lys Gly Ser Phe Ser Ile Gln Tyr Thr Tyr His Val Asp Ala Asn Asp His Asp Gln Ser Arg ~ys Ser Gln Gln Tyr Asp Leu Asn Ala Leu His Lys Thr Thr Lys Ser ~ln Arg His Leu Gly Gly Ser Gln Gln Leu Leu His Asn Lys Gln Glu Gly Arg Asp His Asp Lys Ser Lys Gly His Phe His Arg Val Val Ile His His Lys Gly Gly Lys Ala His Arg G1~ Thr Gln Asn Pro Ser Gln Asp Gln Gly Asn Ser Pro Ser Gly Lys Gly Ile Ser Ser Gln Tyr Ser ~sn Thr Glu Glu Arg Leu Trp Val His Gly Leu Ser Lys Glu Gln Thr ~er Val Ser Gly Ala Gln Lys Gly Arg Ly~ Gln Gly Gly Ser Gln Ser Ser Tyr Val Leu Gln Thr Glu Glu Leu Vz Ala Asn Lys Gln Gln Arg Glu Thr Lys Asn Ser His Gln Asn Lys Gly His Tyr Gln Asn Val Val Glu Val Arg Glu Glu His Ser Ser Lys v21 Gln Thr Ser Leu Cys Pro Ala His Gln Asp Lys Leu Gln His Gly Ser Lys Asp Ile Phe Ser Thr CA 02234763 l998-04-l~

W O 97/14416 PCTAUS96/l6490 > Gln Asp Glu Leu Leu Val Tyr Asn Lys Asn Gln His Gln Thr Lys Asn Leu Asn Gln Asp Gln Gln His Gly Arg Lys Ala Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr Glu Glu Arg Arg Leu His Tyr Gly Glu Asn Gly Val Gln Lys Asp Val Ser Gln Ser Ser Ile Tyr Ser Gln Thr Glu Glu Lys Ala Gln Gly Lys Ser Gln Lys Gln Ile Thr Ile Pro Ser Gln Glu Gln Glu His Ser Gln Lys Ala Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr Glu Glu Arg Arg Leu His Tyr Gly Glu Asn Gly Val Gln Lys Asp Val Ser Gln Arg Ser Ile Tyr Ser Gln Thr Glu Lys Leu Val Ala Gly Lys Ser Gln Ile Gln Ala Pro Asn Pro Lys Gln Glu Pro Trp His Gly Glu Asn Ala Lys Gly Glu Ser Gly Gln Ser Thr Asn Arg Glu Gln Asp Leu Leu Ser His Glu Gln Lys Gly Arg His Gln His Gly Ser His Gly Gly Leu Asp Ile Val Ile Ile Glu Gln Glu Asp Asp Ser Asp Arg His 435 440 *45 Leu Ala Gln His Leu Asn Asn Asp Arg Asn Pro Leu Phe Thr (2) INFORMATION FOR SEQ ID NO:2:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 15 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal CA 02234763 1998-04-1~

WO 97/14416 PCT~US96/16490 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:
Gly Lys Gly Ile Ser Ser Gln Tyr Ser Asn Thr Glu Glu Arg Leu l 5 l0 15 (2) INFORMATION FOR SEQ ID NO:3:
(i) ~u~:N~: CHARACTERISTICS:
(A) LENGTH: ll amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:
Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:4:
(i) ~u~ CHARACTERISTICS:
(A) LENGTH: l9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (Xi) ~QU~N~ DESCRIPTION: SEQ ID No:4:
Gly Glu Asn Gly Val Gln Lys Asp Val Se Gln Arg Ser Ile Tyr Ser l 5 l0 15 Gln Thr Glu CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 ~ (2) INFORMATION FOR SEQ ID NO:5:
_ (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l9 amino acids (B) TYPE: amino acid (C) STR~Mn~n~F..~S: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:
Gly Glu Asn Gly Val Gln Lys Asp Val Ser Gln Ser Ser Ile Tyr Ser l 5 l0 15 Gln Thr Glu (2) INFORMATION FOR SEQ ID NO:6:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 25 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) 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 l 5 l0 15 Arg Arg Leu His Tyr Gly Glu Asn Gly (2) INFORMATION FOR SEQ ID NO:7:

CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 8 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:
Ser Tyr Gln Ser Ser Ser Thr Glu l 5 (2) INFORMATION FOR SEQ ID NO:8:
(i) ~QU~ CHARACTERISTICS:
(A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) sTR~Nn~n-N~.~s: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~QU~N~ DESCRIPTION: SEQ ID NO:8:
Ile Ser Tyr Gln Ser Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:9:
( i ) ~QU~N~ CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) sTR~NnFnN~s: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
- (v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:
Lys Ile Ser Tyr Gln Ser Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:10:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 10 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D~ TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:
Asn Lys Ile Ser Tyr Gln Ser Ser Ser Th-1 5 lQ
(2) INFORMATION FOR SEQ ID NO:11:
(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal CA 02234763 l998-04-l~

W O 97/14416 PCT~US96/16490 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:
Ala Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:12:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 8 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:
Ala Gly Pro Thr Gly Ala Ser Ala (2) INFORMATION FOR SEQ ID NO:13:
(i) ~u~N~ CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) sTR~Nn~N~s single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:
Asn Lys Ile Ser Tyr Gln Ser (2) INFORMATION FOR SEQ ID NO:14:

CA 02234763 l998-04-l~

W O 97/14416 PCT~US96/16490 ~Q~N~ CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single - (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:
Lys Ile Ser Tyr Gln Ser (2) INFORMATION FOR SEQ ID NO:15:
( i ) ~U~N~'~ CHARACTERISTICS:
(A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 12 (D) OTHER INFORMATION: /note= "any natural amino acid"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:
Gly Glu Asn Gly Val Gln Lys Asp Val Ser Gln Xaa Ser Ile Tyr Ser - Gln Thr Glu (2) INFORMATION FOR SEQ ID NO:16:

~ U~N~ CHARACTERISTICS:

CA 02234763 1998-04-1~

(A) LENGTH: 8 amino acids ~B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l':
Asn Lys Ile Ser Tyr Gln Ser Ser l 5 (2) INFORMATION FOR SEQ ID NO:17:
(i) ~Qu~N~ CHARACTERISTICS:
(A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~:Qu~ DESCRIPTION: SEQ ID NO:17:
Asn Lys Ile Ser Tyr Gln Ser Ser Ser l 5 (2) INFORMATION FOR SEQ ID NO:l8:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: lû amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO

-(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Qu~N~ DESCRIPTION: SEQ ID NO:18:
Ala Asn Lys Ile Ser Tyr Gln Ser Ser Ser l 5 l0 (2) INFORMATION FOR SEQ ID NO:l9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Qu~ DESCRIPTION: SEQ ID NO:l9:
Lys Ile Ser Tyr Gln Ser Ser Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:20:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 17 amino acids (B) TYPE: amino acid (C) STR~Mn~nN~ss single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal CA 02234763 1998-04-1~

(xi) ~u~N~: DESCRIPTION: SEQ ID NO:20:
Gln Leu Asp Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr ~is Gln Ser l 5 l0 15 Ser (2) INFORMATION FOR SEQ ID NO:2l:
( i ) ~yU~N~ CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Q~N~ DESCRIPTION: SEQ ID NO:21:
Asn Arg Ile Ser Tyr Gln Ser l 5 (2) INFORMATION FOR SEQ ID NO:22:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (Xi) ~QU~N-~ DESCRIPTION: SEQ ID NO:22:
Asn Lys Val Ser Tyr Gln Ser l 5 CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 , (2) INFORMATION FOR SEQ ID NO:23:
_ (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 8 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:
Asn Lys Met Ser Tyr Gln Ser Ser l 5 (2) INFORMATION FOR SEQ ID NO:24:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 8 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:
Asn Lys Leu Ser Tyr Gln Ser Ser l 5 (2) INFORMATION FOR SEQ ID NO:25:
- (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear CA 02234763 1998-04-1~

(ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:
Asn Lys Ile Thr Tyr Gln Ser Ser Ser (2) INFORMATION FOR SEQ ID NO:26:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single _ (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:
Asn Lys Ile Ser Phe Gln Ser Ser Ser (2) INFORMATION FOR SEQ ID NO:27:
(i) ~-u~N~ CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal PC~nUS96/16490 - g3 -(xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:
~ Asn Lys Ile Ser Trp Gln Ser Ser Ser Thr l 5 l0 (2) INFORMATION FOR SEQ ID NO:28:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: lO amino acids (B) TYPE: amino acid (C~ STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:
Asn Lys Ile Ser Tyr Asn Ser Ser Ser Thr l 5 l0 (2) INFORMATION FOR SEQ ID No:29:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Qu~N~ DESCRIPTION: SEQ ID NO:29:
Asn Lys Ile Ser Tyr Gln Thr Ser Ser Thr l 5 lO
(2) INFORMATION FOR SEQ ID NO:30:
(i) ~u~ CHARACTERISTICS:

CA 02234763 1998-04-1~

(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:
Asn Lys Ile Ser Tyr Gln Ser l 5 (2) INFORMATION FOR SEQ ID NO:3l:
- (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 8 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:
Gln Lys Ile Ser Tyr Gln Ser Ser l 5 (2) INFORMATION FOR SEQ ID NO:32:
( i ) ~U ~:N~ CHARACTERISTICS:
(A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO

CA 02234763 1998-04-1~

(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:
Asn Arg Ile Thr Tyr Gln Ser Ser Ser l 5 (2) INFORMATION FOR SEQ ID NO:33:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C~ STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:
Asn Arg Ile Ser Phe Gln Ser Ser Ser Thr l 5 l0 (2) INFORMATION FOR SEQ ID NO:34:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal -W O 97tl4416 PCTtUS96/16490 (Xi) ~QU~N~ DESCRIPTION: SEQ ID No:34:
Gln Lys Ile Ser Tyr Gln Thr Ser Ser Thr l 5 l0 (2) INFORMATION FOR SEQ ID NO:35:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:
Asn Arg Ile Ser Trp Gln Ser Ser Ser Thr l 5 l0 (2) INFORMATION FOR SEQ ID NO:36:
(i) ~QU~:N~r: CHARACTERISTICS:
(A) LENGTH: lO amino acids (B) TYPE: amino acid ( C ) STR~ N I )1- 1 )N l~ S: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~ u~N~ DESCRIPTION: SEQ ID NO:36:
Asn Ary Ile Ser Tyr Gln Thr Ser Ser Thr l 5 l0 (2) INFORMATION FOR SEQ ID NO:37:
( i ) ~ ~:Q~ : CHARACTERISTICS:

CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 _ - 87 -(A) LENGTH: lO amino acids (B) TYPE: amino acid (C) STR~NnF.nN~.~S: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:
Asn Lys Ile Thr Tyr Gln Thr Ser Ser Th-l 5 l0 (2) INFORMATION FOR SEQ ID NO:38:
- (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) STR~NDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal ~xi) ~QU~N~ DESCRIPTION: SEQ ID NO:38:
Asn Lys Leu Ser Tyr Gln Thr Ser Ser Thr l 5 l0 (2) INFORMATION FOR SEQ ID NO:39:
(i) ~Qu~ CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid - (C) STR~NnFnN~S single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO

CA 02234763 1998-04-1~

- 88 ~

(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:
Gln Lys Leu Ser Tyr Gln Ser Ser Ser Thr l 5 l0 (2) INFORMATION FOR SEQ ID NO:40:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide ~iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:
Asn Arg Leu Ser Tyr Gln Thr Ser Ser Thr l 5 l0 (2) INFORMATION FOR SEQ ID NO:41:
(i) ~U~N~ CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) sTRANnEnN~s single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 (xi) ~QU~:N~ DESCRIPTION: SEQ ID NO:41:
Asn Lys Val Ser Phe Gln Ser Ser Ser Thr l 5 lO
(2) INFORMATION FOR SEQ ID NO:42:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:
Asn Arg Val Ser Trp Gln Ser Ser Ser Thr l 5 l0 (2) INFORMATION FOR SEQ ID NO:43:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) sTR~Mn~n~F~s: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~u~ DESCRIPTION: SEQ ID NO:43:
Gln Lys Val Ser Tyr Gln Ser Ser Ser Thr l 5 l0 (2) INFORMATION FOR SEQ ID NO:44:
( i ) ~U~N~L CHARACTERISTICS:

CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 ._ (A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) sTR~N~nN~ss single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:
Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr l 5 l0 (2) INFORMATION FOR SEQ ID NO:45:
- (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: pep~ide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:
Gly Glu Gln Gly Val Gln Lys Asp Val Ser Gln Ser Ser Ile Tyr Ser l 5 l0 15 Gln Thr Glu (2) INFORMATION FOR SEQ ID NO:46:
(i) ~Q~ CHARACTERISTICS:
(A) LENGTH: 15 amino acids (B) TYPE: amino acid (C) STR~N~ ss: single (D) TOPOLOGY: linear CA 02234763 1998-04-1~

(ii) MOLECULE TYPE: peptide _(iii) HYPOTHETICAL: NO
-(iv) ANTI-SENSE: NO
(v) 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 (2) INFORMATION FOR SEQ ID NO:47:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) sTRANn~nN~s: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:
Gly Glu Asn Gly Leu Gln Lys Asp Val Ser Gln Ser Ser Ile Tyr Ser Gln Thr Glu (2) INFORMATION FOR SEQ ID NO:48:
(i) ~-U~N~ CHARACTERISTICS:
(A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) ~Y~O~ CAL: NO

(iv) ANTI-SENSE: NO

W O 97/14416 PCT~US96/16490 (v) FRAGMENT TYPE: internal (xi) ~Q~N~ DESCRIPTION: SEQ ID No:48:
Gly Glu Asn Gly Val Asn Lys Asp Val Ser Gln Ser Ser Ile Tyr Ser Gln Thr Glu (2) INFORMATION FOR SEQ ID NO:49:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~U~N~ DESCRIPTION: SEQ ID NO:4g:
Gly Glu Asn Gly Val Gln Arg Asp Val Ser Gln Arg Ser Ile Tyr Ser Gln Thr Glu (2) INFORMATION FOR SEQ ID NO:50:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal CA 02234763 l998-04-l5 WO 97/14416 PCT~US96/16490 ,.
-(xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:
Gly Glu Asn Gly Val Gln Lys Asp Val Ser Gln Lys Ser Ile Tyr Ser Gln Thr Glu (2) INFORMATION FOR SEQ ID NO:51:
(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) 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 Gln Thr Glu (2) INFORMATION FOR SEQ ID NO:52:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) STR~NnFnNF~S single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide ~ (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Q~ DESCRIPTION: SEQ ID NO:52:
Gly Glu Asn Gly Val Gln Lys Asp Val Ser Gln Ser Ser Ile Phe Ser l 5 l0 15 Gln Thr Glu (2) INFORMATION FOR SEQ ID NO:53:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:
Gly Glu Asn Gly Val Gln Lys Asp ~et Se- Gln Ser Ser Ile Tyr Thr l 5 l0 15 Gln Thr Glu (2) INFORMATION FOR SEQ ID NO:54:
UU~ CHARACTERISTICS:
(A) LENGTH: l9 amino acids (B) TYPE: amino acid (C) sTR~n~nN~s single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGNENT TYPE: internal (xi) ~Q~N~ DESCRIPTION: SEQ ID NO:54:
Gly Glu Asn Gly Val Gln Lys Asp Val Ser Gln Arg Ser Ile Tyr Thr l 5 l0 15 Gln Thr G1u -CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 _ 95 _ (2) INFORMATION FOR SEQ ID NO:55:
- (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Q~N~: DESCRIPTION: SEQ ID NO:55:
Gly Glu Asn Gly Val Gln Lys Asp Val Ser Gln Ser Ser Ile Tyr Ser Gln Ser Glu (2) INFORMATION FOR SEQ ID No:56:
(i) ~:~U~N~ CHARACTERISTICS:
(A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) 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 15 Asn Thr Glu (2) INFORMATION FOR SEQ ID NO:57:

CA 02234763 l998-04-l~

W O 97/14416 PCTAUS96/l6490 - 96 - t (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 15 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv~ ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:
Gly Lys Ala Ile Ser Ser Gln Tyr Ser Asn Thr Glu Glu Arg Leu (2) INFORMATION FOR SEQ ID NO:58:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 15 amino acids (B) TYPE: amino acid (C) sTRANn~n~s single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~QU~N~: DESCRIPTION: SEQ ID No:58:
Gly Lys Gly Ile Ser Ser Gln Tyr Ser Asn Ser Glu Glu Arg Leu (2) INFORMATION FOR SEQ ID NO:59:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 15 amino acids (B) TYPE: amino acid (C) sTRA~nFnNF~s single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide CA 02234763 1998-04-1~

~ (iii) HYPOTHETICAL: NO
_(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~ UkN~h DESCRIPTION: SEQ ID NO:59:
Gly Arg Gly Ile Ser Ser Gln Tyr Ser Asn Thr Glu Glu Arg Leu l 5 l0 15 (2) INFORMATION FOR SEQ ID NO:60:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 15 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:
Gly Lys Gly Ile Thr Ser Gln Tyr Ser Asn Thr Glu Glu Arg Leu l 5 l0 15 (2) INFORMATION FOR SEQ ID NO:6l:
(i) ~yu~N~ CHARACTERISTICS:
(A) LENGTH: 15 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
-(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~yU~N~ DESCRIPTION: SEQ ID NO:61:

CA 02234763 l998-04-l5 W O 97/14416 PCT~US96/16490 _ 9~ _ Gly Lys Gly Ile Ser Thr Gln Tyr Ser Asn Thr Glu Glu Arg Leu (2) INFORMATION FOR SEQ ID NO:62:
(i) ~Q~N-~ CHARACTERISTICS:
(A) LENGTH: 15 amino acids (B) TYPE: amino acid (C) sTR~Nn~nN~ single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal ~ (xi) ~Q~ DESCRIPTION: SEQ ID NO:62:
Gly Lys Gly Ile Ser Ser Asn Tyr Ser Asn Thr Glu Glu Arg Leu (2) INFORMATION FOR SEQ ID NO:63:
(i) ~QU~N~ CHARACTERISTICS:
(A) LENGTH: 15 amino acids (B) TYPE: amino acid (C) sTRAN~nN~s single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) 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 (2) INFORMATION FOR SEQ ID NO:6~:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 15 amino acids CA 02234763 1998-04-1~

W O 97/14416 PCTrUS96/16490 _ 99 _ (B) TYPE: amino acid (C) STRANDEDNESS: single _ (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) 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 (2) INFORMATION FOR SEQ ID NO:65:
(i) SEQUENCE CHARACTERISTICS:
~ (A) LENGTH: 15 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:
Gly Lys Gly Ile Ser Ser Gln Tyr Thr Asn Ser Glu Glu Arg Leu (2) INFORMATION FOR SEQ ID NO:66:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 15 amino acids (B) TYPE: amino acid (C) sTRANn~nN~s: single - (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO

CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 (iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:
Gly Lys Gly Ile Ser Ser Gln Tyr Ser Asn Ser Glu Glu Arg Leu l 5 l0 15 (2) INFORMATION FOR SEQ ID NO:67:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 25 amino acids (B) TYPE: amino acid (C) sTRANn~nN~s: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67:
Ser Gln Lys Ala Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr Glu Glu l 5 lG 15 Arg Arg Leu His Tyr Gly Glu Asn Gly (2) INFORMATION FOR SEQ ID NO:68:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 8 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 - (xi) ~Qu~ DESCRIPTION: SEQ ID NO:68:
Ile Ser Tyr Gln Ser Ser Ser Thr l 5 (2) INFORMATION FOR SEQ ID NO:69:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: sinyle (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~QU~N~ DESCRIPTION: SEQ ID NO:69:
Ala Asn Lys Ile Ser Tyr Gln Ser Ser (2) INFORMATION FOR SEQ ID NO:70:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
~ (v) FRAGMENT TYPE: internal - (xi) ~yu~ DESCRIPTION: SEQ ID NO:70:
Ala Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr Leu l 5 l0 (2) INFORMATION FOR SEQ ID NO:7l:

W O 97/14416 PCT~US96/16490 U~ CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STR~Nn~nN~.~s: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7l:
Ala Asn Gly Ile Ser Tyr Gln Ser Ser Ser Thr Glu l 5 l0 (2)-INFORMATION FOR SEQ ID No:72:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~U~N~ DESCRIPTION: SEQ ID NO:72:
Ala Asn Pro Ile Ser Tyr Gln Ser Ser Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID No:73:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRAN~:~N~SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide .

,(iii) HYPOTHETICAL: NO
-(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:
Ala Asn Lys Ile Ser Tyr Gln Ser Ala Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:74:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRAN~:~N~:SS: single (D) TOPOLOGY: linear _(ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~QU~N~: DESCRIPTION: SEQ ID NO:74:
Ala Asn Lys Ile Ser Tyr Gln Ser Ser Lys Thr Glu (2) INFORMATION FOR SEQ ID NO:75:
:Qu~ CHARACTERISTICS:
(A) LENGTH: 11 amino acids (B) TYPE: amino acid (C) sTR~Nn~nN~s: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
-(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal -CA 02234763 1998-04-1~

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:75:
Ala Asn Lys Ile Ser Tyr Gln Ser Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:76:
(i) ~QU~ CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) sTR~Nn~nN~ss: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/REY: Peptide (B) LOCATION: 5 (D) OTHER INFORMATION: /label= d-serine /note= "unnatural configuration of the amino acid~

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:76:
Ala Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID No:77:
(i) ~yU~N~: CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) sTRANn~n~s single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICA~: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide CA 02234763 1998-04-1~

(B) LOCATION: 4 (D) OTHER INFORMATION: /label= d-isoleucine /note= "unnatural amino acid stereochemical configuration~

(xi) SEQUENCE DESCRIPTION: SEQ ID No:77:
Ala Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:78:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRANDBDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Q~N~ DESCRIPTION: SEQ ID NO:78:
Ala Asn Lys Ile Ser Tyr Gln Ser Ser Gln Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:79:
Qu~N~ CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHE~ICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:79:
Ala Asn Lys Ile Ser Tyr Gln Ser Ala Lys Thr Glu l 5 l0 CA 02234763 l998-04-l~

W O 97/14416 PCT~US96/16490 (2) INFORMATION FOR SEQ ID NO:80:
(i) ~Qu~ CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRPN~ S: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 3 (D) OTHER INFORMATION: /label= d-lysine /note= "unnatural amino acid stereochemical con~iguration"

(xi) ~UU~N~ DESCRIPTION: SEQ ID NO:80:
Ala Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:81:
(i) ~Q~ CHARACTERISTICS:
(A) LENGTH: 10 amino acids (B) TYPE: amino acid (C) sTRANn~nN~s: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Qu~ DESCRIPTION: SEQ ID NO:81:
Ala Asn Lys Ile Ser Tyr Gln Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:82:

CA 02234763 1998-04-1~

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l0 amino acids _ (B) TYPE: amino acid (C) STRANn~nMF~ single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:82:
Ala Asn Lys Ser Tyr Gln Ser Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:83:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:83:
Ala Asn Lys Ile Tyr Gln Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:84:
(i) ~Qu~ CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) sTR~Nn~nM~s single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide CA 02234763 1998-04-1~

~iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:84:
Ala Asn Lys Ala Ser Tyr Gln Ser Ala Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:85:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid ( C ) STRAN~ ~Fc:s: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~yU~N~ DESCRIPTION: SEQ ID NO:85:
Ala Asn Glu Ile Ser Tyr Gln Ser Ala Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:86:
(i) ~yu~ CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~OU~ DESCRIPTION: SEQ ID NO:86:
Lys Ile Ser Tyr Gln Ser Ser l 5 (2) INFORMATION FOR SEQ ID NO:87:
~i) ~u~:N~ CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:87:
Ser Tyr Gln Ser Ser Thr Glu l 5 (2) INFORMATION FOR SEQ ID NO:88:
(i) ~Q~N~ CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STR~ :.SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~u~ DESCRIPTION: SEQ ID NO:88:
Ser Tyr Gln Ser Ser Thr Leu l 5 (2) INFORMATION FOR SEQ ID NO:89:

CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 8 amino acids (B) TYPE: amino acid (C) STRA~v~N~S: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:89:
Ala Ser Tyr Gln Ser Ser Thr Glu l 5 (2) INFORMATION FOR SEQ ID NO:90:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:90:
Glu Ile Ser Tyr Gln Ser Ser Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:9l:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide CA 02234763 1998-04-1~

WO 97/14416 PCT~US96/16490 -(iii) HYPOTHETICAL: NO
_(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9l:
Ala Asn Glu Ile Ser Tyr Gln Ser Ser Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:92:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Qu~N~ DESCRIPTION: SEQ ID NO:92:
Ala Asn Lys Ile Ser Tyr Tyr Ser Ser Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:93:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal CA 02234763 1998-04-1~

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:93:
Ala Asn Lys Ile Ser Tyr Tyr Ser Ala Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:94:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:94:
Ala Ser Tyr Gln Ser Ser Leu l 5 (2) INFORMATION FOR SEQ ID NO:95:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: l0 amino acids (B) TYPE: amino acid (C) STRA~n~n~s: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) ~Y~O'~ CAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~QU~N~ DESCRIPTION: SEQ ID NO:95:
Ala Asn Ser Tyr Gln Ser Ser Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:96:

--W O 97/14416 PCT~US96/16490 ~QU~N~ ~' CHARACTERISTICS:
(A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:96:
Ala Ser Tyr Gln Ser Ser Ser Thr Glu l 5 (2) INFORMATION FOR SEQ ID NO:97:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 8 amino acids (B) TYPE: amino acid (C) sTRA~nFnMF~s: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) ~Y~O~ CAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Q~ N~ DESCRIPTION: SEQ ID NO:97:
Ser Tyr Gln Ser Ser Ser Thr Glu l 5 (2) INFORMATION FOR SEQ ID NO:98:
(i) ~:QU~N~ CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide WO 97/14416 PCT~US96/16490 (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Q~N~ DESCRIPTION: SEQ ID NO:98:
Ala Asn Lys Ala Ser Tyr Gln Ser Ala Ser Thr Cys l 5 l0 (2) INFORMATION FOR SEQ ID NO:99:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) STRA~n~nN~.SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:99:
Gln Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:l00:
(i) ~Qu~N~ CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANn~nN~S single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) ~Y~O~l'~l'lCAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal CA 02234763 1998-04-1~

, (xi) SEQUENCE DESCRIPTION: SEQ ID NO:100:
Tyr Gln Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:101:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:101:
Ser Gln Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:102:
(i) ~Q~:N~: CHARACTERISTICS:
(A) LENGTH: 10 amino acids (B) TYPE: amino acid (C) sTR~Nn~N~s single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:102:
Ala Asn Lys Ile Ser Gln Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:103:

CA 02234763 l998-04-l5 (i) ~yu~N~ CHARACTERISTICS:
(A) LENGTH: 11 amino acids (B) TYPE: amino acid (C) sTRANn~nN~ss single (D) TOPOLOGY: linear (ii) MOLECU~E TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 3 (~) OTHER INFORMATION: /label= unnatural /note= "ornithine"

(xi) s~Qu~N~: DESCRIPTION: SEQ ID NO:103:
Ala Asn Xaa Ile Ser Tyr Gln Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:104:
(i) ~yu~N~ CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) ~Y~O~ CAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /label= unna~ural /note= "3,4-dichloroph~n~l~n~n~

(xi) ~Q~ DESCRIPTION: SEQ ID NO:104:
Ser Xaa Gln Ser Ser Thr Glu CA 02234763 l998-04-l~

(2) INFORMATION FOR SEQ ID NO:105:
(i) ~Q~ CHAR~CTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /label= unnatural /note= "(3-pyridinyl)alanine"

(xi) ~QU~N~ DESCRIPTION: SEQ ID NO:105:
Ser Xaa Gln Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:106:
(i) ~Qu~ CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) sTR~Nn~nM~s: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:106:
Ser Lys Gln Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:107:
(i) ~Qu~ CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid CA 02234763 1998-04-1~
W O 97/14416 PCT~US96/16490 (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:107:
Ser Tyr Gln Ser Ser Ser Leu l 5 (2) INFORMATION FOR SEQ ID NO:l08:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids _ (B) TYPE: amino acid (C) STR~ N~S: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: l (D) OTHER INFORMATION: /label= unnatural /note= "epsilon aminocaproic acid"

(xi) ~u~ ~ DESCRIPTION: SEQ ID NO:108:
Xaa Tyr Gln Ser Ser Ser Leu l 5 (2) INFORMATION FOR SEQ ID NO:l09:
(i) ~Qu~ CHARACTERISTICS:
(A) LENGTH: ll amino acids (B) TYPE: amino acid ~C) STRANDEDNESS: single (D) TOPOLOGY: linear CA 02234763 1998-04-1~

(ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 4 (D) OTHER INFORMATION: /label= unnatural /note= "N-methylisoleucine"

(xi) ~QU~N~ DESCRIPTION: SEQ ID NO:l09:
Ala Asn Lys Xaa Ser Tyr Gln Ser Ser Thr Glu l 5 l0 (2) INFORMATION FOR SEQ ID NO:ll0:
_ (i) ~QU~N~ CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRA~ ~SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Qukw~ DESCRIPTION: SEQ ID NO:ll0:
Ser Tyr Gln Ser Ser Thr Glu l 5 (2) INFORMATION FOR SEQ ID NO:lll:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRAN~N~SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO

CA 02234763 l998-04-l~

WO 97/14416 PCT~US96/16490 (iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:111:
Tyr Gln Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:112:
(i) SEQUENCE CHAP~ACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI~SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:112:
Ser Tyr Lys Ser Ser Thr Glu l 5 (2) INFORMATION FOR SEQ ID NO:113:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B~ TYPE: amino acid (C) STRANDEDNESS: sin~le (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:113:

CA 02234763 l998-04-l~

Ser Tyr Tyr Ser Ser Thr Glu (2) INFORMATION FOR SEQ ID NO:114:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:114:
Ser Tyr Gln Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:115:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~OU~N~: DESCRIPTION: SEQ ID NO:115:
Ser Tyr Gln Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:116:
(i) ~Qu~N~: CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid CA 02234763 l998-04-l~

W O 97/14416 PCTrUS96/16490 ..

(C) sTR~Nn~nNF~s single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide (B~ LOCATION: 1 (D) OTHER INFORMATION: /label= unnatural /note= "2,3-diaminopropionic acid"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:116:
Xaa Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:117:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 11 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:117:
Ala Asn Lys Ile Ser Tyr Gln Ser Ser Ser Thr (2) INFORMATION FOR SEQ ID NO:118:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRAN~ss: single (D) TOPOLOGY: linear CA 02234763 1998-04-1~

,(ii) MOLECULE TYPE: peptide r( iii ) HYPOTHETICAL: NO
-(iv) ANTI-sENsE: NO
(v) FRAGMENT TYPE: internal (xi) ~OU~ DESCRIPTION: SEQ ID NO:118:
Ala Asn Lys Ala Ser Tyr Gln Ser Ala Ser Thr Leu l 5 l0 (2) INFORMATION FOR SEQ ID NO:ll9:
(i) ~:QU~N~: CHARACTERISTICS:
(A) LENGTH: ll amino acids (B) TYPE: amino acid (C) STRAN~:~N~SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~OU~N~ DESCRIPTION: SEQ ID NO:ll9:
Ala Asn Lys Ala Ser Tyr Gln Ser Ala Ser Leu l 5 l0 (2) INFORMATION FOR SEQ ID NO:120:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ll amino acids (B) TYPE: amino acid -(C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal CA 02234763 l998-04-l~

WO 97/l4416 PCTAUS96/l6490 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:120:
Ala Asn Lys Ala Ser Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:121:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 10 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:121:
Ala Asn Lys Ala Ser Tyr Gln Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:122:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /label= d-leucine /note= "unnatural amino acid stereochemical con~iguration"

CA 02234763 l998-04-l~

WO 97/14416 PCTnJS96/16490 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:122:
Ser Tyr Gln Ser Ser Thr Leu (2) INFORMATION FOR SEQ ID NO:123:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 11 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:123:
Ala Asn Lys Ala Ser Tyr Ala Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:124:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:124:
Lys Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:125:
Qu~: CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid CA 02234763 l998-04-l~

(C) STRAN~:~N~SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:125:
Ser Tyr Gln Ser Ser Lys Leu (2) INFORMATION FOR SEQ ID NO:126:
(i) ~QU~N~ CHARACTERISTICS:
(A) LENGTH: 7 amino acids ~ (B) TYPE: amino acid (C) STRA~V:~N~SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /label= d-leucine /note= "unnatural amino acid stereochemical con~iguration"

(xi) ~ U~N~: DESCRIPTION: SEQ ID NO:126:
Ser Tyr Gln Ser Ser Lys Leu (2) INFORMATION FOR SEQ ID NO:127:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear CA 02234763 1998-04-1~

WO 97/14416 PCT~US96/16490 (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~U~N~ DESCRIPTION: SEQ ID NO:127:
Asn Lys Ile Ser Tyr Tyr Ser (2) INFORMATION FOR SEQ ID NO:128:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) sTRANn~nN~s single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (Xi) ~:~U~:N~'~ DESCRIPTION: SEQ ID NO:128:
Asn Lys Ala Ser Tyr Gln Ser (2) INFORMATION FOR SEQ ID NO:129:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) sTRANnF~nN~s: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal CA 02234763 l998-04-l~

W O 97/14416 PCTrUS96/16490 (xi) ~Qu~N~ DESCRIPTION: SEQ ID NO:129:
Ser Tyr Gln Ser Ser (2) INFORMATION FOR SEQ ID NO:130:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 8 amino acids (B) TYPE: amino acid (C) STR~NDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) ~Y~Ol~h~llCAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:130:
Asn Lys Ile Ser Tyr Gln Ser Ala (2) INFORMATION FOR SEQ ID NO:131:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 8 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:131:
Ala Asn Lys Ile Ser Tyr Tyr Ser CA 02234763 l998-04-l~

WO 97/14416 PCT~US96/16490 (2) INFORMATION FOR SEQ ID NO:132:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 8 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:132:
Ala Asn Lys Ala Ser Tyr Gln Ser (2) INFORMATION FOR SEQ ID NO:133:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:133:
Ser Tyr Gln Ser Ser Thr (2) INFORMATION FOR SEQ ID NO:134:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRPNn~nN~S single (D) TOPOLOGY: linear CA 02234763 l998-04-l~

W O 97/14416 PCT~US96/16490 (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13~:
Ser Tyr Gln Ser Ser Ser (2) INFORMATION FOR SEQ ID NO:135:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:135:
Ser Tyr Gln Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:136:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) sTRAN~nN~s single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal CA 02234763 l998-04-l5 (xi) ~QU~N~ DESCRIPTION: SEQ ID NO:136:
Ala Asn Lys Ile Ser Tyr Gln Ser Ala (2) INFORMATION FOR SEQ ID NO:137:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:137:
Ala Asn Lys Ile Ser Tyr Tyr Ser Ser (2) INFORMATION FOR SEQ ID NO:138:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~u~ DESCRIPTION: SEQ ID NO:138:
Ala Asn Lys Ile Ser Tyr Tyr Ser Ala CA 02234763 1998-04-1~

(2) INFORMATION FOR SEQ ID NO:139:
(i) ~Qu~ CHARACTERISTICS:
(A) LENGTH: 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:139:
Ala Asn Lys Ala Ser Tyr Gln Ser Ala (2) INFORMATION FOR SEQ ID NO:140:
(i) ~:Qu~N~ CHARACTERISTICS:
(A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~yU~N~: DESCRIPTION: SEQ ID NO:140:
Lys Tyr Gln Ser Ser (2) INFORMATION FOR SEQ ID NO:141:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) STR~Nn~nN~.~S: single (D) TOPOLOGY: linear CA 02234763 l998-04-l~

WO 97/14416 PCT~US96/16490 (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
_ (iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /label= homoarginine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:141:
Xaa Tyr Gln Ser Ser (2) INFORMATION FOR SEQ ID NO:142:
(i) SEQUENCE CHARACTERISTICS:
- (A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Qu~N~ DESCRIPTION: SEQ ID NO:142:
Lys Tyr Gln Ser Ser Ser (2) INFORMATION FOR SEQ ID NO:143:
(i) ~QU~N~'~ CHARACTERISTICS:
A (A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO

CA 02234763 l998-04-l~

W O 97/14416 PCT~US96/16490 ~iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide (B~ LOCATION: 1 (D) OTHER INFORMATION: /label= homoarginine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:143:
Xaa Tyr Gln Ser Ser Ser (2) INFORMATION FOR SEQ ID NO:144:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (xi) ~Q~:~ DESCRIPTION: SEQ ID NO:144:
Ser Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:145:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal CA 02234763 l998-04-l~

t (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /label= homoarginine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:145:
Xaa Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:146:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(v) FRAGMENT TYPE: internal (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /label= norleucine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:146:
Lys Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:147:

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= "cyclohexyl~l ~n;n~"

CA 02234763 l998-04-l~
W O 97/14416 PCT~US96/16490 (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 5 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:147:
Xaa Xaa Gln Ser Leu (2) INFORMATION FOR SEQ ID NO:148:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/REY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product=~"homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= ~homotyrosine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 6 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:148:
Xaa Xaa Gln Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:149:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear CA 02234763 l998-04-l~

W 0 97/14416 PCT~US96/16490 (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= "cyclohexylhomoalanine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 6 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:149:
Xaa Xaa Gln Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:150:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 10 amino acids (B) TYPE: amino acid (C) STRAN~N~SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 10 (D) OTHER INFORMATION: /product= '~cyclohexylalanine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:150:
Ala Asn Lys Ala Ser Tyr Gln Ser Ser Xaa (2) INFORMATION FOR SEQ ID NO:151:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide CA 02234763 l998-04-l~
W O 97/14416 PCT~US96/16490 (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:151:
Xaa Tyr Gln Ser Ser Pro (2) INFORMATION FOR SEQ ID NO:152:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iX) FEATURE:
(A) NAME/REY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:152:
Xaa Tyr Gln Ser Ser His (2) INFORMATION FOR SEQ ID NO:153:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= ''homoarginine' (xi) SEQUENCE DESCRIPTION: SEQ ID NO:153:
Xaa Tyr Gln Ser Asn CA 02234763 l998-04-l~

WO 97/14416 PCT~US96/16490 (2) INFORMATION FOR SEQ ID NO:154:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANv~N~SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:154:
Xaa Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:155:
(i) ~Qu~ CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product=
"4-aminomethylphenylalanine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 - (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:155:
Xaa Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:156:

CA 02234763 l998-04-l~
WO 97/14416 PCT~US96/16490 (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:156:
Xaa Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:157:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 10 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 10 (D) OTHER INFORMATION: /product= "cyclohexylalanine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:157:
Ala Asn Lys Ala Lys Tyr Gln Ser Ser Xaa (2) INFORMATION FOR SEQ ID NO:158:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:

CA 02234763 l998-04-l~

(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product=
"2(4,6-dimethylpyrimidine)lysine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:158:
Xaa Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:159:
(i) ~Qu~N~ CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "N'-(2-imidazolyl)lysine"

(xi) ~:Qu~ DESCRIPTION: SEQ ID NO:159:
Xaa Tyr Gln Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:160:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 10 amino acids (B) TYPE: amino acid (C) STRA~N~SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 5 (D) OTHER INFORMATION: /product= ~homoarginine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:160:

CA 02234763 l998-04-l~
W 0 97/14416 PCT~US96/l6490 Ala Asn Lys Ala Xaa Tyr Gln Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:161:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product=
"(4-~minocyclohexyl)alanine (ix) FEATURE:
(A) NAME/KEY: Peptide _ (B) LOCATION: 7 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:161:
Xaa Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:162:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STR~MD~nM~.~S: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "N'-(2-imidazolyl)lysine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /product= ~'norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:162:

CA 02234763 1998-04-1~

W O 97/14416 PCT~US96/16490 Xaa Tyr Gln Ser Ser Ser Leu l 5 (2) INFORMATION FOR SEQ ID NO:163:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: l (D) OTHER INFORMATION: /product= "homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= "cyclohexylalanine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:l63:
Xaa Xaa Gln Ser Ser Ser Leu l 5 (2) INFORMATION FOR SEQ ID NO:164:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
- (A) NAME/KEY: Peptide (B) LOCATION: l (D) OTHER INFORMATION: /product= ~'homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /product= "homoarginine"

CA 02234763 1998-04-1~
W O 97/14416 PCTtUS96tl6490 (xi) ~Ou~N~: DESCRIPTION: SEQ ID NO:164:
Xaa Tyr Gln Ser Ser Ser Xaa (2) INFORMATION FOR SEQ ID NO:165:
(i) ~Qu~:N~: CHARACTERISTICS:
(A) LENGTH: lû amino acids tB) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (~) LOCATION: 5 (D) OTHER INFORMATION: /product= "N'-(2-imidazolyl)lysine"
(ix) FEATURE:
_ (A) NAME/KEY: Peptide (B) LOCATION: 10 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:165:
Ala Asn Lys Ala Xaa Tyr Gln Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:166:
(i) ~Ou~ CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= ~3-iodotyrosine"
(ix) FEATURE:
(A) NAME/KEY: Peptide --CA 02234763 1998-04-1~

WO 97/14416 PCT~US96/16490 (B) LOCATION: 7 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:l66:
Xaa Xaa Gln Ser Ser Ser Leu l 5 (2) INFoRMATIoN FOR SEQ ID NO:167:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: l _ (D) OTHER INFORMATION: /product= "homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product=
"O-dimethylphosphotyrosine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /product= "norleuci~e"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:167:
Xaa Xaa Gln Ser Ser Ser Leu l 5 (2) INFORMATION FOR SEQ ID NO:l68:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid - (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: l _ CA 02234763 l998-04-l~
W O 97/14416 PCT~US96/16490 (D) OTHER INFORMATION: /product= "homoarginine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:168:
Xaa Tyr Gln Ser Ser Asp (2) INFORMATION FOR SEQ ID NO:169:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= ~homoarginine~
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= "O-methyltyrosine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /product= ~'norleucine~

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:169:
Xaa Xaa Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:170:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 10 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 10 (D) OTHER INFORMATION: /product= ~norleucine"

, CA 02234763 l998-04-l~

WO 97/14416 PCTAJS96/l6490 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:170:
Ala Asn Lys Ala Lys Tyr Gln Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:171:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRAN~N~SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"
(ix) FEATURE:
_ (A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= "cyclohexylalanine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:171:
Xaa Xaa Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:172:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGT~: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= 'IN'-(2-imidazolyl)lysine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= ~'cyclohexyl~l~n;ne"

(ix) FEATURE:
(A) NAME/KEY: Peptide CA 02234763 l998-04-l~

(B) LOCATION: 7 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:172:
Xaa Xaa Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:173:
(i) ~Q~N~ CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRAN~N~SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 _ (D) OTHER INFORMATION: /product= "homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= 'Icyclohexylalanine'' (xi) SEQUENCE DESCRIPTION: SEQ ID NO:173:
Xaa Xaa Gln Ser Ser Ser (2) INFORMATION FOR SEQ ID NO:174:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 ~D) OTHER INFORMATION: /product= "homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= "cyclohexylalanine"

, CA 02234763 l998-04-l~

(ix) FEATURE:
(A) NAMB/KEY: Peptide (B) LOCATION: 6 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:174:
Xaa Xaa Gln Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:175:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix~ FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= ~homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= "cyclohexylalanine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 6 (D) OTHER INFORMATION: /product= ~norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:175:
Xaa Xaa Gln Ser Pro Leu (2) INFORMATION FOR SEQ ID NO:176:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid ' (C) STRANDEDNESS: single A ( D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:

CA 02234763 l998-04-l~
WO 97/14416 PCT~US96/16490 (A) NAMEtKEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= "3-~luorotyrosine'~
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:176:
Xaa Xaa Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:177:
(i) SEQUENCE CHARACTERISTICS:
_ (A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) STRA~v:v~SS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:177:
Xaa Tyr Gln Ser Pro (2) INFORMATION FOR SEQ ID NO:178:
(i) SEQUENCE CHAR~CTERISTICS:
(A) LENGTH: 6 amino acids tB) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 6 CA 02234763 l998-04-l~

(D) OTHER INFORMATION: /product= ~norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:178:
Lys Tyr Gln Ser Lys Leu (2) INFORMATION FOR SEQ ID NO:179:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= "~-aminophenylalanine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /product= "norleucine"

(xi) ~;Qu~;N~:~; DESCRIPTION: SEQ ID NO:179:
Xaa Xaa Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:180:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide ~.
(ix) FEATURE.
(A) NAME/KEY: Peptide (B) LOCATION:
(D) OTHER INFORMATION: /product= "homoarginine"

CA 02234763 l998-04-l~

(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product=
"7-HO-tetrahydroiso~uinoline CO2H"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:180:
Xaa Xaa Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:181:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single _ (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "ornithine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:181:
Xaa Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:182:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide CA 02234763 l998-04-l~

~ (xi) SEQUENCE DESCRIPTION: SEQ ID NO:182:
Lys Ala Ala Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:183:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 7 (D) OTHER INFORMATION: /product= ~norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:183:
Lys Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:184:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 11 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:184:
Leu Asn Lys Ala Ser Tyr Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:185:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide CA 02234763 1998-04-1~
W O 97/14416 PCTtUS96tl6490 (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 2 (D) OTHER INFORMATION: /product= "cyclohexylalanine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:185:
Xaa Xaa Gln Ser Ser (2) INFORMATION FOR SEQ ID NO:186:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) STRA~N~SS: single _ (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:186:
Tyr Gln Ser Ser (2) INFORMATION FOR SEQ ID NO:187:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:187:
Xaa Tyr Gln Ser CA 02234763 l998-04-l~

WO 97/14416 PCT~US96/16490 ..
(2) INFORMATION FOR SEQ ID NO:188:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "homoarginine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:188:
Xaa Tyr Gln Ser Ser Ser (2) INFORMATION FOR SEQ ID NO:189:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 5 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 5 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:189:
Gln Ser Ser Ser Leu v (2) INFORMATION FOR SEQ ID NO:190:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide CA 02234763 l998-04-l~
WO 97/14416 PCTnJS96/l6490 (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product=
"7-HO-tetrahydro-3-iso~uinoline CO2H"
(ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 6 (D) OTHER INFORMATION: /product= "norleucine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:190:
Xaa Gln Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:l91:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 11 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single _ (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l91:
Ala Asn Lys Ala Ser Tyr Ala Ser Ser Ser Leu (2) INFORMATION FOR SEQ ID NO:192:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:192:
Ser Tyr Gln Ser Ser Lys Leu (2) INFORMATION FOR SEQ ID NO:193:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 9 amino acids (B) TYPE: amino acid , CA 02234763 l998-04-l5 W O 97/14416 PCT~US96/16490 (C) STRAN~N~SS: single ~ (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) S~U~N~ DESCRIPTION: SEQ ID NO:193:
Ala Asn Lys Ala Ser Tyr Gln Ser Leu (2) INFORMATION FOR SEQ ID NO:194:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids ~B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Peptide (B) LOCATION: 1 (D) OTHER INFORMATION: /product= "ornithine"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:194:
Xaa Tyr Gln Ser Ser Ser Leu

Claims (28)

WHAT IS CLAIMED IS:

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, i) cyclophosphamide, 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) mitomycin C, i) porfiromycin, j) 5-fluorouracil, k) 6-mercaptopurine, l) cytosine arabinoside, m) podophyllotoxin, n) etoposide, o) etoposide phosphate, p) melphalan, q) vinblastine, r) vincristine, s) leurosidine, t) vindesine, u) estramustine, v) cisplatin, w) cyclophosphamide, x) leurosine, and y) taxol;

or the pharmaceutically acceptable salt thereof.

5. 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 5 wherein the conjugate is of the formula I:

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-tetrahydroisoquinoline-3-carboxylic acid;
R is hydrogen or -(C=O)R1; and R1 is C1-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) AsnLysIleSerTyrGln~Ser (SEQ.ID.NO.: 13), b) LysIleSerTyrGln~Ser (SEQ.ID.NO.: 14), c) GlyGluAsnGlyValGlnLysAspValSerGlnXaaSerIleTyr~SerGlnThrGlu (SEQ.ID.NO.: 15), d) GlyLysGlyIleSerSerGlnTyr~SerAsnThrGluGluArgLeu (SEQ.ID.NO.: 2), e) AsnLysIleSerTyrTyr~Ser (SEQ.ID.NO.: 127), f) AsnLysAlaSerTyrGln~Ser (SEQ.ID.NO.: 128), g) SerTyrGln~SerSer (SEQ.ID.NO.: 129), h) LysTyrGln~SerSer (SEQ.ID.NO.: 140);
i) hArgTyrGln~SerSer (SEQ.ID.NO.: 141);
j) hArgChaGln~SerSer (SEQ.ID.NO.: 185); and k) TyrGln~SerSer (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:

wherein X is:
AsnLyslleSerTyrGlnSer-- (SEQ.ID.NO.: 13), AsnLyslleSerTyrGlnSerSer-- (SEQ.ID.NO.: 16), AsnLyslleSerTyrGlnSerSerSer-- (SEQ.ID.NO. 17), AsnLyslleSerTyrGlnSerSerSerThr-- (SEQ.ID.NO.:10), AsnLyslleSerTyrGlnSerSerSerThrGlu-- (SEQ.ID.NO.: 3), AlaAsnLyslleSerTyrGlnSerSerSerThrGlu-- (SEQ.ID.NO.: 11), N-terminus Ac-AlaAsnLyslleSerTyrGlnSerSerSerThr- (SEQ.ID.NO.: 117), Ac-AlaAsnLyslleSerTyrGlnSerSerSerThrLeu- (SEQ.ID.NO.: 70), Ac-AlaAsnLysAlaSerTyrGlnSerAlaSerThrLeu- (SEQ.ID.NO.: 118), Ac-AlaAsnLysAlaSerTyrGlnSerAlaSerLeu- (SEQ.ID.NO.: 119), Ac-AlaAsnLysAlaSerTyrGlnSerSerSerLeu- (SEQ.ID.NO.: 120), Ac-AlaAsnLysAlaSerTyrGlnSerSerLeu- (SEQ.ID.NO.: 121), Ac-SerTyrGlnSerSerSerLeu- (SEQ.ID.NO.: 144), Ac-hArgTyrGlnSerSerSerLeu- (SEQ.ID.NO.: 145), Ac-LysTyrGlnSerSerSerLeu- (SEQ.ID.NO.: 124), or Ac-LysTyrGlnSerSerNle- (SEQ.ID.NO.: 146), N-terminus or the pharmaceutically acceptable salt thereof.

10. The method of treatment according to Claim 7 wherein the conjugate is selected from:

Ac-hArgTyraln-SerSerPro-dox(3') (SEQ.ID.NO. : 151) Ac-hArgTyrGln-SerPro-dox(3') (SEQ.ID.NO.: 177) Ac-hArgTyrGln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 154) Ac-AmfTyrGln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 155) H2NCO-hArgTyrGln-SerSerSerLeu-dox(3') (SEQ.ID.NO.: 156) Ac-LysTyrGln-SerSerNle-dox(3') (SEQ.ID.NO.: 146) Ac-LysTyrGln-SerLysNle-dox(3') (SEQ.ID.NO.: 178) Ac(cis-p-NH2Cha)TyrGlnSerSerNledox(3') (SEQ.ID.NO.: 161) Ac-AlaAspLysAla(hArg)TyrGln-SerSerLeu-dox(3') (SEQ.ID.NO.: 160) Ac-hArgTyrGln-SerAsn-dox(3') (SEQ.ID.NO.: 153) Ac-hArgTyrGln-SerSerHis-dox(3') (SEQ.ID.NO.: 152) Ac-(imidazolyl)LysTyrGln-SerSerLeu-dox(3') (SEQ.ID.NO.: 159) Ac-(imidazolyl)LysTyrGlnSerSerSerNle-dox(3') (SEQ.ID.NO.: 162) Ac-hArg(Cha)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 163) Ac-hArg(Me2P03Tyr)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 167) Ac-hArgTyrGln-SerSerSerhArg-dox(3') (SEQ.ID.NO.: 164) Ac-hArg(3-Iodo-Tyr)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 166) Ac-hArg(O-Me-Tyr)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 169) Ac-hArg(p-NH2-Phe)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 179) Ac-hArg(Cha)Gln-SerSerNle-dox(3') (SEQ.ID.NO.: 174) Ac-hArg(Cha)Gln-SerProNle-dox(3') (SEQ.ID.NO.: 175) Ac(imidazolyl)Lys(Cha)GlnSerSerSerNle-dox(3') (SEQ.ID.NO.: 172) Ac-hArg(7-HO-TIC)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 180) Ac-hArg(3-Fluoro)TyrGlnSerSerSerNle-dox(3') (SEQ.ID.NO.: 176) Ac-(ornithine)TyrGln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 181) Ac-LysAlaAlaSerSerSerLeu-dox(3') (SEQ.ID.NO.: 183) Ac-hArgh(Cha)Gln-SerSerNle-dox(3') (SEQ.ID.NO.: 149) Ac-AlaArgLysAlaSerTyrGln-SerLeu-dox(3') (SEQ.ID.NO.: 193) and Ac-(Orn)TyrGln-SerSerSerLeu-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 II:

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-tetrahydroisoquinoline-3-carboxylic acid; or XL is - NH- (CH2)n - NH -R is hydrogen or-(C=O)R1;
R1 is C1-C6-aLkyl or aryl;

R19 is hydrogen or acetyl; and n is 1,2,3,4 or 5, or the pharmaceutically acceptable salt thereof.

12. The method of treatment according to Claim 6 wherein the conjugate is of the formula IIa:

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, or the pharmaceutically acceptable salt thereof.

13. The method of treatment according to Claim 11 wherein the conjugate is selected from:

and or the pharmaceutically acceptable salt thereof.

14. 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 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.

15. 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 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. The method of treatment according to Claim 15 wherein the conjugate is or the pharmaceutically acceptable salt thereof.

17. 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.

18. 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, or the pharmaceutically acceptable salt thereof.

19. The composition according to Claim 18 wherein the conjugate is of the formula I:

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-tetrahydroisoquinoline-3-carboxylic acid;

R is hydrogen or -(C=O)R1; and R1 is C1-C6-alkyl or aryl, or the pharmaceutically acceptable salt thereof.

20. The composition according to Claim 18 wherein the conjugate is selected from:

wherein X is:

AsnLyslleSerTyrGlnSer-- (SEQ.ID.NO.: 13), AsnLyslleSerTyrGlnSerSer-- (SEQ.ID.NO.: 16), AsnLyslleSerTyrGlnSerSerSer-- (SEQ.ID.NO.: 17), AsnLyslleSerTyrGlnSerSerSerThr-- (SEQ.ID.NO.:10), AsnLyslleSerTyrGlnSerSerSerThrGlu-- (SEQ.ID.NO.: 3), AlaAsnLyslleSerTyrGlnSerSerSerThrGlu-- (SEQ.ID.NO.: 11), N-terminus Ac~AlaAsnLysIleSerTyrGlnSerSerSerThr~ (SEQ.ID.NO.: 117), Ac~AlaAsnLysIleSerTyrGlnSerSerSerThrLeu~ (SEQ.ID.NO.: 70), Ac~AlaAsnLysAlaSerTyrGlnSerAlaSerThrLeu~ (SEQ.ID.NO.: 118), Ac~AlaAsnLysAlaSerTyrGlnSerAlaSerLeu~ (SEQ.ID.NO.: 119), Ac~AlaAsnLysAlaSerTyrGlnSerSerSerLeu~ (SEQ.ID.NO.: 120), Ac~AlaAsnLysAlaSerTyrGlnSerSerLeu~ (SEQ.ID.NO.: 121), Ac~SerTyrGlnSerSerSerLeu~ (SEQ.ID.NO.: 144), Ac~hArgTyrGlnSerSerSerLeu~ (SEQ.ID.NO.: 145), Ac~LysTyrGlnSerSerSerLeu~ (SEQ.ID.NO.: 124), or or the pharmaceutically acceptable salt thereof.

21. The composition according to Claim 18 wherein the conjugate is selected from:
Ac-hArgTyrGln-SerSerPro-dox(3') (SEQ.ID.NO.: 151) Ac-hArgTyrGln-SerPro-dox(3') (SEQ.ID.NO.: 177) Ac-hArgTyrGln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 154) Ac-AmfTyrGln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 155) H2NCO-hArgTyrGln-SerSerSerLeu-dox(3') (SEQ.ID.NO.: 156) Ac-LysTyrGln-SerSerNle-dox(3') (SEQ.ID.NO.: 146) Ac-LysTyrGln-SerLysNle-dox(3') (SEQ.ID.NO.: 178) Ac(cis-p-NH2Cha)TyrGlnSerSerNledox(3') (SEQ.ID.NO.: 161) Ac-AlaAspLysAla(hArg)TyrGln-SerSerLeu-dox(3') (SEQ.ID.NO.: 160) Ac-hArgTyrGln-SerAsn-dox(3') (SEQ.ID.NO.: 153) Ac-hArgTyrGln-SerSerHis-dox(3') (SEQ.ID.NO.: 152) Ac-(imidazolyl)LysTyrGln-SerSerLeu-dox(3') (SEQ.ID.NO.: 159) Ac-(imidazolyl)LysTyrGlnSerSerSerNle-dox(3') (SEQ.ID.NO.: 162) Ac-hArg(Cha)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 163) Ac-hArg(Me2PO3Tyr)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 167) Ac-hArgTyrGln-SerSerSerhArg-dox(3') (SEQ.ID.NO.: 164) Ac-hArg(3-Iodo-Tyr)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 166) Ac-hArg(O-Me-Tyr)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 169) Ac-hArg(p-NH2-Phe)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 179) Ac-hArg(Cha)Gln-SerSerNle-dox(3') (SEQ.ID.NO.: 174) Ac-hArg(Cha)Gln-SerProNle-dox(3') (SEQ.ID.NO.: 175) Ac(imidazolyl)Lys(Cha)GlnSerSerSerNle-dox(3') (SEQ.ID.NO.: 172) Ac-hArg(7-HO-TIC)Gln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 180) Ac-hArg(3-Fluoro)TyrGlnSerSerSerNle-dox(3') (SEQ.ID.NO.: 176) Ac-(ornithine)TyrGln-SerSerSerNle-dox(3') (SEQ.ID.NO.: 181) Ac-LysAlaAlaSerSerSerLeu-dox(3') (SEQ.ID.NO.: 183) Ac-hArgh(Cha)Gln-SerSerNle-dox(3') (SEQ.ID.NO.: 149) Ac-AlaArgLysAlaSerTyrGln-SerLeu-dox(3') (SEQ.ID.NO.: 193) and Ac-(Orn)TyrGln-SerSerSerLeu-dox(3') (SEQ.ID.NO.: 194) or the pharmaceutically acceptable salt thereof.

22. The composition according to Claim 18 wherein the conjugate is of the formula II:

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-tetrahydroisoquinoline-3-carboxylic acid; or XL is -NH-(CH2)n-NH-R is hydrogen or-(C=O)R1;
R1 is C1-C6-aLkyl or aryl;
R19 is hydrogen or acetyl; and n is 1, 2, 3, 4 or 5, or the pharmaceutically acceptable salt thereof.

23. The composition according to Claim 22 wherein the conjugate is:

or the pharmaceutically acceptable salt thereof.

24. The composition according to Claim 18 wherein the conjugate is of the formula II:

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, or the pharmaceutically acceptable salt thereof.

25. The composition according to Claim 18 wherein the conjugate is:

or the pharmaceutically acceptable salt thereof.

26. A pharmaceutical composition useful for treating an 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, 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.

27. 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 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.

28. The composition according to Claim 27 wherein the conjugate is or the pharmaceutically acceptable salt thereof.