PEPTIDES IN CANCER RESEARCH Cancer Research

PEPTIDE-BASED CANCER THERAPEUTICS

This brochure discusses the potential use of as anti- cancer drugs highlighting current scenario and future prospects. Some peptides are also used as diagnostic tools for cancer detection. G-protein-coupled receptors are most important targets in drug development. Many of them are overexpressed in tumor cells. Amongst them, the GnRH receptor is the target of a consider- able number of GnRH agonists and antagonists used in cancer management. GnRH (-releasing hormone) or LHRH (-releasing hormone) is a decapeptide pro- duced in in the hypothalamus and released in a pulsatile fashion into the pituitary portal circulation. Prolonged non-pulsatile administration of LHRH leads to down-regulation of LH and FSH secretion, followed by a suppression of gonadal steroid synthesis. For this reason, longer-acting GnRH agonists as well as antago- nists are used for the treatment of hormone-dependent breast and prostate cancers. Most neuroendocrine tumors show a marked overexpression of receptors, especially of sst2, which instigated the development of somatostatin agonists as . These compounds also play an important role in diagnosis. Bombesin/gastrin-releasing receptors can be overex- pressed in malignant cells. Antagonists of these peptides inhibit tumor growth. Active immunization by peptide vaccines is another promising strategy to fight cancer.

2 Introduction penetrate tumor tissue. Their proteolytic Cancer is characterized by uncontrolled degradation can be conveniently prevented division of cells and the ability of these by chemical modifications such as incor- cells to invade other tissues leading to the poration of D-amino acids or cyclization. formation of tumor mass, to vasculariza- Properties of bicyclic peptides are even tion and, finally, to metastasis (spread of better and comparable to those of antibody cancer to other parts of the body). Though drugs. angiogenesis (growth of new blood vessels The peptide drugs currently available on the from existing vessels) is a normal and vital market can be classified as analogs and process during growth and development, it antagonists of peptide hormones or tumor is also a fundamental step in the transition targeting agents carrying radionuclides. of tumors from a dormant state to a malig- nant one. So, angiogenesis inhibitors have LHRH (GnRH) Agonists and Antagonists been used to suppress tumor cell growth. The first example for the introduction of Chemotherapy is one of the classical ap- peptide drugs into cancer therapy is the proaches to treat cancer, a cytotoxic agent use of LHRH (luteinizing hormone-releasing is delivered to the cancer cells. The main hormone) analogs. Schally et al. developed problem with conventional chemotherapy the first GnRH agonists which later were is its inability to administer the correct applied in the treatment of prostate and amount of drug directly to cancer cells with- breast cancer. Since then, peptides such as out affecting normal cells. Drug resistance, altered biodistribution, biotransformation and premature clearance are also common problems. Targeted chemotherapy and drug MODIFICATION AT delivery techniques are emerging as a pow- erful method to circumvent such problems. This will allow the selective and effec- POSITION 6 WITH tive localization of drugs at pre-defined targets (e.g. overexpressed receptors) while A D-AMINO ACID restricting its access to normal cell thus maximizing therapeutic index and reduc- ing toxicity. The discovery of further recep- YIELDS POTENT tors abnormally expressed in cancer cells and tumor-related peptides and proteins is expected to lead to a ‘new wave’ of more LONG-ACTING effective and selective anti-cancer drugs in the future. LHRH AGONISTS The “biologics” approach to cancer therapy includes application of proteins, monoclo- nal antibodies and peptides. Monoclonal , leuprolide, , , antibodies (mAb) and large protein ligands and have been developed and have two major limitations compared to approved in cancer therapy. Depot formu- peptides: poor delivery to tumors due to lations of these peptides allow for a more their large size and a dose-limiting toxicity efficacious and convenient treatment of pa- in liver and bone marrow due to nonspecific tients with . Administration uptake into the reticuloendothelial system. of these peptides effects a down-regulation The use of such macromolecules has there- of GnRH receptors in the pituitary, leading fore been restricted to vascular targets to an inhibition of follicle-stimulating hor- present on the luminal side of the tumor mone (FSH) and LH release, and a concomi- vessel endothelium and to hematological tant decrease in testosterone production. malignancies. The introduction of LHRH antagonists as Peptides possess many advantages such as resulted in therapeutic improve- small size, ease of synthesis and modi- ment over agonists as they cause an imme- fication, they are biocompatible and can diate and dose-related inhibition of LH and

3 Cancer Research

PEPTIDE SEQUENCE INDICATION GnRH Pyr-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH₂ none in cancer therapy GnRH Agonists BUSERELIN Pyr-His-Trp-Ser-Tyr-D-Ser(tBu)-Leu-Arg-Pro-NHEt Prostate cancer GOSERELIN Pyr-His-Trp-Ser-Tyr-D-Ser(tBu)-Leu-Arg-Pro-Azagly-NH₂ Prostate and breast cancer HISTRELIN Pyr-His-Trp-Ser-Tyr-D-His(Bzl)-Leu-Arg-Pro-NHEt Prostate and breast cancer LEUPROLIDE Pyr-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt Prostate and breast cancer TRIPTORELIN Pyr-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH₂ Prostate and breast cancer GnRH Antagonists Ac-D-2-Nal-D-4-Cpa-D-3-Pal-Ser-N-Me-Tyr-D-Asn- Prostate cancer Leu-Lys(isopropyl)-Pro-D-Ala-NH₂ CETRORELIX Ac-D-2-Nal-4-chloro-D-Phe-β-(3-pyridyl)-D-Ala-Ser- Prostate and breast cancer Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH₂ DEGARELX Ac-D-2-Nal-D-4-Cpa-D-3-Pal-Ser-4-amino-Phe(L-4,5- Prostate cancer dihydroorotyl)-4-ureido-D-Phe-Leu-Lys(isopropyl)-Pro- D-Ala-NH₂ OZARELIX Ac-D-2-Nal-D-4-Cpa-D-3-Pal-Ser-N-Me-Tyr-D-Hci- Prostate cancer Nle-Arg-Pro-D-Ala-NH₂ TEVERELIX Ac-D-2-Nal-D-4-Cpa-D-3-Pal-Ser-Tyr-D-Hci-Leu- Prostate cancer Lys(isopropyl)-Pro-D-Ala-NH₂

Table1. LHRH agonists and new generation antagonists available in the market.

FSH by competitive blockade of the LHRH Most neuroendocrine tumors (NETs) feature receptors. To date, many potent GnRH an- a strong overexpression of somatostatin tagonists are available for therapeutic use receptors, mainly of subtype 2 (sst2). Cur- in patients suffering from prostate cancer. A rently, five subtypes list of such agonists and antagonists avail- (sst) are known (sst1-5). The density of able in the market can be found in Table 1 these receptors on tumor tissue is vastly higher than on healthy tissue. Therefore, Somatostatin Analogs in Cancer Therapy sst are attractive targets for delivery of Apart from the use of peptidic LHRH ago- radionuclides employing appropriately nists and antagonists for treating cancer, modified somatostatin analogs. Introduced somatostatin analogs are the only approved in the late 1980s by Sandoz, [111In-DTPA]- cancer therapeutic peptides in the market. octreotide (pentetreotide, Octreoscan®), Potent agonists of somatostatin (SRIF) in- rapidly became the gold standard for cluding octreotide (sandostatin) have been diagnosis of sst-positive NETs. Numer- developed for the treatment of acromegaly, ous peptide-based tumor-imaging agents gigantism and thyrotropinoma associated targeting sst have been developed over the with syndrome, and diarrhea in past decades. Octreoscan® and NeoTect® patients with vasoactive intestinal peptide- (technetium-99m-labeled depreotide,

secreting tumors (VIPomas). , cyclo(MePhe-Tyr-D-Trp-Lys-Val-Hcy(CH2CO-

another long-acting analog of somatostatin, β-Dap-Lys-Cys-Lys-NH2)) are the only is used in the management of acromegaly radiopeptide tracers on the market ap- and symptoms caused by neuroendocrine proved by the FDA. An octreotide scan or tumors. octreoscan is a scintigraphic method used

4 THE FIVE KNOWN SOMATOSTATIN RECEPTORS ARE ATTRACTIVE TARGETS FOR TUMOR DIAGNOSIS AND THERAPY

to find and other types of tumors cancerous cells relies on vaccines consist- and to localize sarcoidosis. DTPA-Octreo- ing of peptides derived from the amino acid tide, after radiolabeling with indium-111, sequence of candidate tumor-associated or is injected into a vein and travels through specific antigens. Tumor cells express anti- the bloodstream. The radioactive octreotide gens known as tumor-associated antigens attaches to tumor cells that have receptors (TAAs) that can be recognized by the T-cells for somatostatin. A radiation-measuring of the host’s immune system. A consider- device detects the radioactive octreotide, able number of TAAs could be identified and and generates images showing the precise characterized. TAAs can be injected into location of the tumor in the body. cancer patients in an attempt to induce a The principle also works in cancer therapy. systemic immune response that may result Peptide receptor radionuclide therapy in the destruction of the cancer cells. Any (PRRT) combines appropriately modified oc- protein/peptide produced in a tumor cell treotide with a radionuclide, which will bind that has mal structure due to mutation to carcinoid tumor cells with overexpressed can act as a tumor antigen. Such abnormal somatostatin receptors. Once bound, the proteins are produced due to mutations targeted radiation will kill the malignant in the corresponding gene. Hence, clinical cells the peptide is bound to. The complex between radionuclide and peptide has to be stable, especially if the radiopeptide is used in therapy. Cyclic chelators as DOTA bind (radio)nuclides as 68Ga, 90Y, or 177Lu more tightly, so (Tyr3)-DOTA- octreotide (DOTATOC, edotreotide) can be used in diagnosis and therapy of NETs. This also holds true for the C-terminal acid, DOTA- (DOTATATE). Somatostatin agonists vary in receptor selectivity: Lanreotide shows high affin- ity for sst2 and somewhat less to sst5. , another SRIF agonist, binds less selectively and thus mimics the natural ligand more closely.

Peptide Vaccines Active immunization seems to be the most Figure1. Different treatment options of cancer using promising strategy to treat cancer though peptides. Peptides can be used as: anti-cancer drug, many approaches based on the employ- cytotoxic drug carrier, vaccine, hormone, radionuclide ment of immune cells or immune molecules carrier and drug target (cancer drugs can be targeted have been followed. This method of treating towards tumor associated peptides or peptide receptors. (J. Thundimadathil, J. Amino Acids 2012, 13 (2012))

5 Cancer Research

studies have been initiated to assess the neuroendocrine cells might be responsible therapeutic potential of active immuniza- for the development and progression of tion or vaccination with TAA peptides in prostate cancer to independence. patients with metastatic cancer. So far, GRP is widely distributed in lung and gas- only a limited number of TAA peptides, trointestinal tracts. It is produced in small mostly those recognized by CD8 (+) T-cells cell lung cancer (SCLC), breast, prostatic, in melanoma patients, have been clinically and pancreatic cancer, and functions as a tested. Several melanoma TAAs have been growth factor. The involvement of bombe- identified and are being evaluated as pep- sin-like peptides in the pathogenesis of a tide-based cancer vaccines in clinical trials wide range of human tumors, their function around the world. as autocrine/paracrine tumoral growth Recent advances in the field of molecular factors, and the high incidence of BN/ biology have enabled the rapid identifica- GRP receptors in various human cancers tion of dozens of candidate TAAs for several prompted the design and synthesis of BN/ important human cancers GRP receptor (GRPR) antagonists such as RC-3095, RC-3940-II, and RC-3950. Current Status and Future of Peptide Currently, many researchers are focus- Based Anti-Cancer Agents ing on the development of GHRH (growth The application of peptides as direct thera- hormone releasing hormone - a hypothal- peutic agents, in targeted drug delivery amic polypeptide) antagonists as potential and as diagnostic tools in cancer biology anti-cancer therapeutics since GHRH is is growing. Among many improvements in produced by various human tumors, includ- targeted and controlled delivery of thera- ing prostate cancer, and seems to exert peutics, specifically binding peptides have an autocrine/paracrine stimulatory effect emerged as the most valuable non-immu- on them. Another promising approach for nogenic approach to target cancer cells. the therapy of prostate cancer consists of Various cancer treatment options using the use of cytotoxic analogues of GnRH, peptides are summarized in Figure 1. bombesin, and somatostatin, which can be The RGD peptide iRGD (CRGDKGPDC) is targeted to receptors for these peptides in able to specifically recognize and penetrate prostate cancers and their metastases. For cancerous tumors but not normal tissues. example, a potential drug candidate, AEZS- The development of similar peptides with 108 consists of a peptide LHRH, coupled to extraordinary tumor-penetrating properties the chemotherapeutic agent doxorubicin to will definitely make substantial improve- directly target cells that express GnRH re- ments in cancer treatment in future. Chloro- ceptors, specifically, prostate cancer cells. toxin (Bachem product 4044876, a 36 amino acid peptide isolated from scorpion venom) There is a tremendous effort to discover has a higher affinity for glioma cells than angiogenesis inhibitors, based on polypep- for non-neoplastic and normal brain cells tides as the safest and least toxic therapy This preferential binding has allowed the for diseases associated with abnormal development of new methods for the treat- angiogenesis. A number of ongoing clini- ment and diagnosis of brain cancer. Anti- cal trials in this area focus on peptides angiogenesis as a therapeutic approach derived from: extracellular matrix proteins, led to renewed interest in cilengitide. This growth factors and growth factor receptors, integrin inhibitor, a cyclic RGD peptide, is coagulation cascade proteins, chemokines, being evaluated as non-small-cell lung can- Type I Thrombospondin domain containing cer therapeutic in clinical trials. proteins and serpins. Bombesin/gastrin-releasing peptide (BN/ GRP) peptides were shown to bind selec- Peptides will make tively to the G-protein–coupled receptors a huge impact in the area of cancer on the cell surface, stimulating the growth diagnosis and of various malignancies in murine and therapy in the near human cancer models. Thus, it has been future. proposed that the secretion of BN/GRP by

6 REFERENCES

L.J. Hofland et al. L. Vujanovic and L.H. Butterfield P. Whelan Somatostatin analogs: clinical appli- Melanoma cancer vaccines and Triptorelin embonate: a 6-month cation in relation to human soma- anti-tumor T cell responses. formulation for prostate cancer. tostatin receptor subtypes. J. Cell. Biochem. 102, 301-310 (2007) Expert Opin. Pharmacother. 11, Biochem. Pharmacol. 50, 287-297 C. Doehn et al. 2929-2932 (2010) (1995) for prostate cancer. D. Wild et al. J.C. Reubi et al. Expert Opin. Investig. Drugs 18, 851- First clinical evidence that imaging Somatostatin receptors and their 860 (2009) with somatostatin receptor antago- subtypes in human tumors and in F. Goel et al. nists is feasible. peritumoral vessels. LHRH agonists for adjuvant therapy J. Nucl. Med. 52, 1412-1417 (2011) Metabolism 45, 39-41 (1996) of early breast cancer in premeno- B.L.R. Kam et al. A.V. Schally pausal women. Lutetium-labelled peptides for Luteinizing hormone-releasing hor- Cochrane Database Syst. Rev. therapy of neuroendocrine tumours. mone analogs: their impact on the CD004562 (2009) Eur. J. Nucl. Med. Mol. Imaging 39 control of tumorigenesis. A. Hackshaw Suppl. 1, S103-112 (2012) Peptides 20, 1247-1262 (1999) Luteinizing hormone-releasing K.P. Koopmans and A.W. Glaude- W.A. Breeman et al. hormone (LHRH) agonists in the mans Somatostatin receptor-mediated treatment of breast cancer. Rationale for the use of radiola- imaging and therapy: basic science, Expert Opin. Pharmacother. 10, belled peptides in diagnosis and current knowledge, limitations and 2633-2639 (2009) therapy. future perspectives. F. Klug et al. Eur. J. Nucl. Med. Mol. Imaging 39 Eur. J. Nucl. Med. 28, 1421-1429 Characterization of MHC ligands for Suppl 1, S4-10 (2012) (2001) peptide based tumor vaccination. P. Limonta et al. S.V. Le et al. Curr. Pharm. Des. 15, 3221-3236 GnRH receptors in cancer: from cell PAC1 and PACAP expression, signal- (2009) biology to novel targeted therapeutic ing, and effect on the growth of G. Mezö and M. Manea strategies. HCT8, human colonic tumor cells. Luteinizing hormone-releasing hor- Endocr. Rev. 33, 784-811 (2012) Regul. Pept. 109, 115-125 (2002) mone antagonists. P.W. Moody et al. C. Borghouts et al. Expert Opin. Ther. Pat. 19, 1771-1785 Pituitary adenylate cyclase-acti- Current strategies for the develop- (2009) vating polypeptide causes ment of peptide-based anti-cancer R.M. Myers et al. phosphorylation of the epidermal therapeutics. Cancer, chemistry, and the cell: growth factor receptor in lung can- J. Pept. Sci. 11, 713-726 (2005) molecules that interact with the cer cells. L.K. Kvols and E.A. Woltering neurotensin receptors. J. Pharmacol. Exp. Ther. 341, 873-881 Role of somatostatin analogs in the ACS Chem. Biol. 4, 503-525 (2009) (2012) clinical management of non-neuro- E.D. Deeks P.K. Nanda et al. endocrine solid tumors. Histrelin: in advanced prostate Bombesin analogues for gastrin- Anticancer Drugs 17, 601-608 (2006) cancer. releasing peptide receptor imaging. R.T. Dorsam and J.S. Gutkind Drugs 70, 623-630 (2010) Nucl. Med. Biol. 39, 461-471 (2012) G-protein-coupled receptors and F. Hohla and A.V. Schally J. Thundimadathil cancer. Targeting gastrin releasing pep- Cancer treatment using peptides: Nat. Rev. Cancer 7, 79-94 (2007) tide receptors: New options for the Current therapies and future pros- S. Fister et al. therapy and diagnosis of cancer. pects. Gonadotropin-releasing hormone Cell Cycle 9, 1738-1741 (2010) J. Amino Acids 2012, 13 (2012) type II antagonists induce apoptotic P.J. Pommerville and J.G. de Boer F. Barbieri et al. cell death in human endometrial GnRH antagonists in the treatment Peptide receptor targeting in cancer: and ovarian cancer cells in vitro and of advanced prostate cancer. the somatostatin paradigm. in vivo. Can. J. Urol. 17, 5063-5070 (2010) Int. J. Pept. 2013, 926295 (2013) Cancer Res. 67, 1750-1756 (2007)

7 Cancer Research

R.A. Feelders et al. A. Accardo et al. B. Tang et al. Pasireotide, a multi-somatostatin Receptor binding peptides for Vasoactive intestinal peptide recep- receptor ligand with potential ef- target-selective delivery of nanopar- tor-based imaging and treatment of ficacy for treatment of pituitary and ticles encapsulated drugs. tumors (Review). neuroendocrine tumors. Int. J. Nanomedicine 9, 1537-1557 Int. J. Oncol. 44, 1023-1031 (2014) Drugs Today (Barc.) 49, 89-103 (2013) (2014) Z. Blumenfeld and A. Evron E. Harford-Wright et al. V. Ambrosini et al. Preserving fertility when choosing The potential for substance P an- The use of -68 labeled soma- chemotherapy regimens - the role tagonists as anti-cancer agents in tostatin receptors in PET/CT imaging. of gonadotropin-releasing hormone brain tumours. PET Clin. 9, 323-329 (2014) agonists. Recent Pat. CNS Drug. Discov. 8, 13- R. Baldelli et al. Expert Opin. Pharmacother. 16, 23 (2013) Somatostatin analogs therapy in 1009-1020 (2015) O. Keskin and S. Yalcin gastroenteropancreatic neuroendo- L. Dardevet et al. A review of the use of somatostatin crine tumors: current aspects and Chlorotoxin: a helpful natural scor- analogs in oncology. new perspectives. pion peptide to diagnose glioma and Onco Targets Ther. 6, 471-483 (2013) Front. Endocrinol. (Lausanne) 5, 7 fight tumor invasion. F.G. Rick et al. (2014) Toxins (Basel) 7, 1079-1101 (2015) Agonists of luteinizing hormone-re- N.J. Carter and S.J. Keam M. Muñoz and R. Coveñas leasing hormone in prostate cancer. Degarelix: a review of its use in pa- Targeting NK-1 Receptors to Prevent Expert Opin. Pharmacother. 14, 2237- tients with prostate cancer. and Treat Pancreatic Cancer: a New 2247 (2013) Drugs 74, 699-712 (2014) Therapeutic Approach. N.D. Shore et al. K. Mander et al. Cancers (Basel) 7, 1215-1232 (2015) New considerations for ADT in Advancing drug therapy for brain Y. Nishimura et al. advanced prostate cancer and the tumours: a current review of the pro- Cancer immunotherapy using novel emerging role of GnRH antagonists. inflammatory peptide Substance P tumor-associated antigenic pep- Prostate Cancer Prostatic. Dis. 16, and its antagonists as anti-cancer tides identified by genome-wide 7-15 (2013) agents. cDNA microarray analyses. U.W. Tunn et al. Recent Pat. CNS Drug Discov. 9, 110- Cancer Sci. 106, 505-511 (2015) Six-month depot 121 (2014) J. Yang et al. formulations in advanced prostate C. Morgat et al. Composite peptide-based vaccines cancer: a clinical evaluation. Targeting neuropeptide receptors for cancer immunotherapy (Review). Clin. Interv. Aging 8, 457-464 (2013) for cancer imaging and therapy: per- Int. J. Mol. Med. 35, 17-23 (2015) R. Varshney et al. spectives with bombesin, neuroten- (68)Ga-labeled bombesin analogs sin, and neuropeptide-Y receptors. for receptor-mediated imaging. J. Nucl. Med. 55, 1650-1657 (2014) Recent Results Cancer Res. 194, 221- J.W. Moul 256 (2013) Utility of LHRH antagonists for ad- Z. Yu et al. vanced prostate cancer. An update of radiolabeled bombesin Can. J. Urol. 21, 22-27 (2014) analogs for gastrin-releasing pep- R. Coveñas and M. Muñoz tide receptor targeting. Cancer progression and substance P. Curr. Pharm. Des. 19, 3329-3341 Histol. Histopathol. 29, 881-890 (2013) (2014) O. Abdel-Rahman et al. M. Muñoz and R. Coveñas Somatostatin receptor expression in Involvement of substance P and the hepatocellular carcinoma: prognos- NK-1 receptor in pancreatic cancer. tic and therapeutic considerations World J. Gastroenterol. 20, 2321- Endocr. Relat. Cancer 21, R485-493 2334 (2014) (2014)

8 PEPTIDES FOR CANCER RESEARCH OFFERED BY BACHEM A choice of our products. Besides peptides, Bachem offers enzyme substrates, inhibitors, amino acid derivatives and other compounds for numerous applications in cancer research.

9 Cancer Research

(Des-Gly10,D-Ser(tBu)6,Pro-NHEt9)- (Des-Gly10,D-His2,D-Ser(tBu)6,Pro- BUSERELIN AND LHRH (Buserelin) NHEt9)-LHRH ((D-His2)-Buserelin) 4028150 4018942 IMPURITIES

(D-Ser(tBu)6,Azagly10)-LHRH (D-Ser4,D-Ser(tBu)6,Azagly10)-LHRH GOSERELIN AND (Goserelin) ((D-Ser4)-Goserelin) IMPURITIES 4038252 4035304

(D-Ser(tBu)6,Azagly10)-LHRH (D-Ser6,Azagly10)-LHRH (Goserelin (free base)) ((D-Ser6)-Goserelin) 4073798 4047922

(D-His2,D-Ser(tBu)6,Azagly10)-LHRH (D-Tyr5,D-Ser(tBu)6,Azagly10)-LHRH ((D-His2)-Goserelin) ((D-Tyr5)-Goserelin) 4035303 4035302

6 7 10 10 6 9 (D-Ser(tBu) ,D-Leu ,Azagly )-LHRH (Des-Gly ,D-Ser(tBu) ,Pro-NHNH2 )- ((D-Leu7)-Goserelin) LHRH ((Des-carboxamide)-Goserelin) 4037082 4039400

(Ser(Ac)4,D-Ser(tBu)6,Azagly10)-LHRH ((Ser(Ac)4)-Goserelin) 4054732

(Ser(tBu)6,Azagly10)-LHRH ((Ser(tBu)6)- Goserelin) 4035307

10 (Des-Gly10,D-His(Bzl)6,Pro-NHEt9)- (Des-Gly10,D-Tyr5,D-His(Bzl)6,Pro- HISTRELIN, LHRH (Histrelin) NHEt9)-LHRH ((D-Tyr5)-Histrelin) 4035442 4028763 ANALOGS AND

FRAGMENTS (Des-Gly10,D-His2,D-His(Bzl)6,Pro- (D-His(Bzl)6)-LHRH (1-7) (free acid) NHEt9)-LHRH ((D-His2)-Histrelin) (Histrelin (1-7)) 4028762 4029428

(Des-Gly10,D-Ser4,D-His(Bzl)6,Pro- NHEt9)-LHRH ((D-Ser4)-Histrelin) 4025431

(Des-Gly10,D-Leu6,Pro-NHEt9)-LHRH (Des-Gly10,D-His2,D-Leu6,Pro-NHEt9)- LEUPROLIDE AND (Leuprolide) LHRH ((D-His2)-Leuprolide) 4033014 4028311 IMPURITIES

10 6 13 7 10 2 4 6 (Des-Gly ,D-Leu ,[ C6]Leu ,Pro- (Des-Gly ,D-His ,D-Ser ,D-Leu ,Pro- NHEt9)-LHRH NHEt9)-LHRH ((D-His2,D-Ser4)- 13 7 (([ C6]Leu )-Leuprolide) Leuprolide) 4048762 4048380 13

((D-Leu6)-LHRH (1-8) (free acid) ((Des- (Des-Gly10,D-Leu6,D-Leu7,Pro-NHEt9)- Pro-NHEt9)-Leuprolide) LHRH ((D-Leu7)-Leuprolide) 4037360 4028310

(D-Leu6,Pro-NHEt9)-LHRH (4-9) (Des-Gly10,D-Leu6,Orn8,Pro-NHEt9)- (Leuprolide (4-9)) LHRH ((Orn8)-Leuprolide) 4027694 4065128 SYlLRP-NHEt

11 Cancer Research

(Des-Gly10,D-Pyr1,D-Leu6,Pro-NHEt9)- (Des-Gly10,Des-Ser4,D-Leu6,Pro- LEUPROLIDE AND LHRH ((D-Pyr1)-Leuprolide) NHEt9)-LHRH ((Des-Ser4)-Leuprolide) 4046113 4065127 IMPURITIES

(CONTINUED) (Des-Gly10,D-Ser4,D-Leu6,Pro-NHEt9)- (Des-Gly10,Leu6,Pro-NHEt9)-LHRH LHRH ((D-Ser4)-Leuprolide) ((Leu6)-Leuprolide) 4033803 4033802

(Des-Gly10,D-Trp3,D-Leu6,Pro-NHEt9)- (Des-Gly10,Ser(Ac)4,D-Leu6,Pro-NHEt9)- LHRH ((D-Trp3)-Leuprolide) LHRH ((Ser(Ac)4)-Leuprolide) 4058815 4034724

(Des-Gly10,D-Tyr5,D-Leu6,Pro-NHEt9)- (Des-Pyr1,Des-Gly10,D-Leu6,Pro- LHRH ((D-Tyr5)-Leuprolide) NHEt9)-LHRH ((Des-Pyr1)-Leuprolide) 4028312 4034726

(D-Trp6)-LHRH (Trp6)-LHRH TRIPTORELIN, (Triptorelin Acetate salt) ((Trp6)-Triptorelin) 4033016 4029115 ANALOGS AND

FRAGMENTS (D-Trp6)-LHRH (D-Trp6)-LHRH-Leu-Arg-Pro-Gly amide (Triptorelin Pamoate salt) 4029116

4039648

4026526

4025364

4025366 For-HWSYwLRPG-NH2

4025363 HWSYwLRPG-NH2

12 Cetrorelix Ozarelix* LHRH 4039760 4040823 Ac-D-2Nal-D-4Cpa-D-3Pal-SY-D-Cit- Ac-D-2Nal-D-4-Cpa-D-3Pal-S-N-Me-Y- ANTAGONISTS LRPa-NH D-Hci-Nle-RPa-NH 2 2

Degarelix* 4047459 Ac-D-2Nal-D-4Cpa-D-3Pal-S-4-amino- Phe(L-4,5-dihydroorotyl)-4-ureido-D- Phe-LK(isopropyl)-Pa-NH₂

Somatostatin-14 Octreotide trifluoroacetate salt SOMATOSTATIN, 4033009 (Dimer, Antiparallel) AGCKNFFWKTFTSC 4080984 AGONISTS (fCFwKTC-ThrO) 2 ([ring-D₅]Phe⁶)-Somatostatin-14 AND 4072026 Octreotide trifluoroacetate salt 2 AGCKNF( H5)FWKTFTSC (Dimer, Parallel) ANTAGONISTS 4080983

BIM-23627 (fCFwKTC-ThrO)2 4041642 F(4Cl)c-2Pal-WKVC-2Nal-NH₂ DOTA-(Tyr³)-Octreotate (DOTATATE) Cyclo-Somatostatin 4046724 (Somatostatin Antagonist) DOTA-fCYwKTCT 4007608 c(7Aha-FwKT(Bzl)) Pasireotide* NEW (SOM230) Lanreotide 4060221 (BIM-23014) c(-Hyp(2-aminoethylcarbamoyl)-Phg- 4067760 wKY(Bzl)F)

D-2Nal-CYwKVCT-NH2 Tyr-(D-Dab⁴,Arg⁵,D-Trp⁸)-cyclo- Octreotide acetate salt Somatostatin-14 (4-11) (SMS 201-995) (KE 108) 4042822 4041674 fCFwKTC-ThrOl Y-c(D-Dab-RFFwKTF)

Octreotide pamoate salt (D-Phe⁵,Cys⁶·¹¹,N-Me-D-Trp⁸)- 4076740 Somatostatin-14 (5-12) amide fCFwKTC-ThrOl ( (N-Me-D-Trp⁴)-Octreotate amide) 4039201 ([ring-D₅]Phe³)-Octreotide fCY(NMe-w)KTCT 4069671 2 fC( H5)FwKTC-ThrO (RC-160) 4062833 fCYwKVCW-NH₂

* Bachem provides this product solely for uses within the scope of any statute or law providing for an immunity, exemption, or exception to patent infringement (“Exempted Uses”), including but not limited to 35 U.S.C. § 271(e)(1) in the United States, the Bolar type exemption in Europe, and any corresponding exception to patent infringement in any other country. It is the sole responsibility of the purchaser or user of this product, and the purchaser or user of this product agrees to engage only in such Exempted Uses, and to comply with all applicable intellectual property laws and/or regulations. The purchaser of this product agrees to indem- nify Bachem against all claims in connection with the performance of the respective commercial agreement (e.g. supply agreement) and possible infringements of intellectual property rights. 13 Cancer Research

Bombesin (Tyr⁴,D-Phe¹²)-Bombesin BOMBESIN AND 4010579 4016626 BOMBESIN/GRP pEQRLGNQWAVGHLM-NH₂ pEQRYGNQWAVGfLM-NH₂ (Lys³)-Bombesin (D-2-Nal⁵,Cys⁶·¹¹,Tyr⁷,D-Trp⁸,Val¹⁰,2- ANTAGONISTS 4011270 Nal¹²)-Somatostatin-14 (5-12) amide pEQKLGNQWAVGHLM-NH₂ (BIM 23042) 4025467 (D-Phe¹²)-Bombesin D-2Nal-CYwKVC-Nal-NH 4030216 pEQRLGNQWAVGfLM-NH₂ (Deamino-Phe¹⁹,D-Ala²⁴,D-Pro²⁶- psi(CH₂NH)Phe²⁷)-GRP (19-27) (human, (D-Phe⁶,Leu-NHEt¹³,des-Met¹⁴)- porcine, canine) Bombesin (6-14) (BW-10, BW2258U89) (DPDMB) 4030653

4030433 Deamino-FHWAVaHpo(Ψ[CH2NH])F-NH₂ fQWAVGHL-NHEt

(D-Phe⁶,Leu¹³-psi(CH₂NH)p-chloro- Phe¹⁴)-Bombesin (6-14) 4030432

fQWAVGHL(Ψ[CH2NH])F(4-Cl)-NH₂ (Tyr⁴)-Bombesin 4011655 pEQRYGNQWAVGHLM-NH₂

Phenylacetyl-(D-Arg²·²⁸,p-chloro- 4031294 GHRH/ Phe⁶,Homoarg⁹·²⁹,Tyr(Me)¹⁰,Abu¹⁵, Rw(MeF)wLM-NH₂₂ Nle²⁷)-GRF (1-29) amide (human) NEUROTENSIN/ (JV-1-38) Ac-Trp-3,5-bis(trifluoromethyl)benzyl 4034002 ester SUBSTANCE P Phac-YrDAIF(4Cl)TN-Har-Y(Me)RKVL- (L-732,138, Substance P Antagonist) Abu-QLSARKLLQDI-Nle-r-Har-NH₂ 4025344

(D-Arg¹,D-Pro²,D-Trp⁷·⁹,Leu¹¹)- PACAP-38 (6-38) (human, chicken, Substance P mouse, ovine, porcine, rat) ((D-Pro2)-Spantide) 4031158 4004685 FTDSYSRYRKQMAVKKYLAAVLG- rpKPQQwFwLL-NH₂ KRYKQRVKNK-NH₂

(D-Arg¹,D-Trp⁵·⁷·⁹,Leu¹¹)-Substance P Acetyl-(D-Phe²,Lys¹⁵,Arg¹⁶,Leu²⁷)-VIP 4031292 (1-7)-GRF (8-27) rPKPwQwFwLL-NH₂ (PG 97–269) 4048647 (Arg⁶,D-Trp⁷·⁹,N-Me-Phe⁸)-Substance D-2Nal-CYw-Orn-VC-2Nal-NH₂ P (6-11) (Antagonist G)

14 Myristoyl-(Lys¹²·²⁷·²⁸)-VIP-Gly-Gly- VIP/PACAP Thr (free acid) 4037992 Myr-HSDAVFTDNYTKLRKQMAVK- KYLNSIKKGGT

VIP Antagonist 4031352 KPRRPYTDNYTRLRKQMAVKKYLN-

SILN-NH2

Ovalbumin (257-264) (chicken) Cytochrome C (88-104) EPITOPES 4033142 (domestic pigeon) SIINFEKL 4044055 KAERADLIAYLKQATAK Ovalbumin (323-339) Collagen Type IV α3 Chain (185-203) (chicken, japanese quail) 4028340 4034255 CNYYSNSYSFWLASLNPER ISQAVHAAHAEINEAGR

Chlorotoxin VARIOUS 4044876 MCMPCFTTDHQMARKCDDCCGGK- GRGKCYGPQCLCR-NH₂

Human CMV pp65 (495-503) 4039108 NLVPMVATV

Z-Val-Ala-DL-Asp-fluoromethyl- ketone 4026865 Z-VAD-FMK

15 Cancer Research

OESOPHAGEAL CANCER

Oesophageal cancer. Light micro- graph of an oesophageal cancer biopsy. This is an epidermoid cancer, a type of cancer that arises from the epithelial cells that line the oeso- phagus (gullet). Left is a large vacuo- le filled with a whorl of the protein keratin. Symptoms of oesophageal cancer may include difficulty swal- lowing, weight loss and vomiting. Risk factors include smoking and alcohol consumption. Treatment is surgical removal of the tumor, often combined with chemotherapy or radiotherapy.

(KEYSTONE/SCIENCE PHOTO LIBRARY)

16 API PRODUCTS Bachem is the world´s leading independent manufacturer of peptide active pharmaceutical ingredients (APIs) and a well established man- ufacturer of small molecules APIs. Each year, Bachem manufactures hundreds of batches of drug substance for projects in clinical trials and for products on the market.

Bachem is currently involved in more than 150 cGMP development projects targeting NCEs and Bachem offers a range of more than 30 generic drug substances. We have the capacity to produce peptide APIs from gram scale up to annual quantities of hundreds of kilograms and small molecules APIs from gram scale up to annual quantities of tens of tons. Our GMP manufacturing facilities are located in Switzer- land and the United States and are regularly inspected by the FDA and local authorities.

In addition to close to 50 years of experience in the manufacture of drug substance, Bachem also has a strong regulatory background and we are well prepared to fully support you with the required regu- latory documentation such as drug master files (DMFs). For complex development projects we support you with dedicated project teams comprising of our experts from R&D, production, quality control, qual- ity assurance and regulatory affairs. A team of experienced Business Development Managers and Generics Managers look forward to work- ing with you for your future requirements.

17 Cancer Research

Buserelin Leuprolide Acetate GENERIC APIs 4028150-GMP 4033014-GMP

Gonadorelin Acetate Triptorelin Acetate 4033013-GMP 4033016-GMP

Goserelin Acetate Triptorelin Pamoate 4038252-GMP 4039648-GMP

Impurity A Impurity F IMPURITIES OF (D-Ser4)-Leuprolide (D-His2,D-Ser4)-Leuprolide 4033803 4048380 THE

LEUPRORELIN Impurity B Impurity G (D-His2)-Leuprolide (D-Tyr5)-Leuprolide PH. EUR. 4028311 4028312 MONOGRAPH

Impurity D Impurity I (Ser(Ac)4)-Leuprolide (D-Pyr1)-Leuprolide 4034724 4046113

Impurity E (D-Trp3)-Leuprolide 4058815

18 Impurity A Impurity G IMPURITIES OF (D-Ser4)-Goserelin (D-His2)-Goserelin 4035304 4035303 THE GOSERELIN

PH. EUR. Impurity B Impurity K (Ser(tBu)6)-Goserelin (Ser(Ac)4)-Goserelin MONOGRAPH 4035307 4054732

Impurity E Impurity L 9 7 (Pro-NHNH2 )-Buserelin (D-Leu )-Goserelin 4039400 4037082

Impurity F (D-Tyr5)-Goserelin 4035302

Impurity A IMPURITIES OF (D-His2)-Buserelin 4018942 THE BUSERELIN

Somatostatin Octreotide Acetate SOMATOSTATIN 4033009-GMP 4042822-GMP AND AGONISTS

* Bachem provides this product solely for uses within the scope of any statute or law providing for an immunity, exemption, or exception to patent infringement (“Exempted Uses”), including but not limited to 35 U.S.C. § 271(e)(1) in the United States, the Bolar type exemption in Europe, and any corresponding exception to patent infringement in any other country. It is the sole responsibility of the purchaser or user of this product, and the purchaser or user of this product agrees to engage only in such Exempted Uses, and to comply with all applicable intellectual property laws and/or regulations. The purchaser of this product agrees to indem- nify Bachem against all claims in connection with the performance of the respective commercial agreement (e.g. supply agreement) and possible infringements of intellectual property rights.

19 Some products may berestricted countries. incertain We cannot bemadeliable for any possible errors ormisprints. All information iscompiled to thebest of ourknowledge. shop.bachem.com or shoponline www.bachem.com Visit ourwebsite [email protected] Tel. +41585952020 Bachem Europe, Africa, Middle East andIndia [email protected] AG Tel. +81366610774 K.K. Bachem Japan Asia Pacific [email protected] +1 3105394171 Tel. +1 8884222436(toll free inUSA &Canada) Bachem Americas, Inc. Americas Marketing &Sales Contact www.bachem.com shop.bachem.com

2012436 published by Global Marketing, Bachem AG, May 2020