Photochemistry 2024

PHOTO CHEMISTRY

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Empowering Peptide Innovation

With this guiding theme in mind, Iris Biotech’s mission is to support researchers by supplying

• innovative technologies, rare compounds, as well as a broad portfolio on standard consumables, available in flexible quantities from small scale to bulk quantities. To fulfill our dedication “Empowering Peptide Innovation”, we are attending various conferences, symposia, and exhibitions each year. This allows us to remain in direct contact with scientists all over the world, both from academia and industry, to exchange knowledge, and to gather new ideas to tackle your current challenges.

Guided by our dedication to provide competent service, as well as novel substances and latest technologies,

Iris Biotech is your trusted partner for the world of peptides, while having strong expertise in associated disciplines. Thus, our portfolio comprises reagents and tools for the synthesis and modification of peptides, e.g. amino acids, resins and solvents but also for related technologies such as Drug Delivery, Linkerology® and Life Sciences.

Acids Building Blocks Life Sciences Drug Delivery Reagents Resins Linkerology® Click Chemistry

Owed to the growing demand for tailor-made compounds, our portfolio is fine-tuned by our Custom Synthesis Service at Iris Biotech Laboratories. Our skilled scientists offer profound expertise in

• de novo route development,

• upscaling towards larger scale production,

• as well as synthesis optimization for increased efficiency.

Examples are the synthesis of rare chiral building blocks, unnatural amino acid derivatives, sophisticated orthogonal protecting groups, heterocycles, building blocks for nucleotides, PEGs and PEG-analogues as well as specific linkers for controlled drug delivery and release.

Amino

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Portfolio Overview

Peptide Synthesis and Modification

(Protected) Amino Acids

Standards such as Fmoc-D/L-AAA and Boc-D/L-AAA, Smoc amino acids for peptide synthesis in water, variety of protecting groups (e.g. Pbf, Trt, tBu, Bzl, Acm, Mob, SIT, Phacm, Allocam, Mmt), unusual amino acids, fluorinated derivatives, substituted prolines, arginine analogues

Building Blocks

Amino alcohols, amino aldehydes, diamines and hydrazines, (pseudoproline) dipeptides, polyamines and spermines, fatty acid derivatives

Reagents

Coupling reagents, solvents and scavengers, protecting groups

Resins

Preloaded resins (e.g. based on Trityl, TCP, TentaGel, Methoxybenzhydryl, Merrifield, PAM, Rink, Wang), scavenger resins, hydrazone resins

Linkerology® and Drug Delivery Life Sciences

Linkers for Solid Phase Peptide Synthesis

Cleavable Linkers

Val-Ala based, Val-Cit based, disulfide-based, Dde-helping hands

Photo-Activatable Linkers

Functionalized Linkers

Clickable linkers, trifunctional linkers, linkers with maleimide function, cross-linkers, selective N-term acylation and biotinylation

PROTACs

Ligands, linkers & modules

Fullerenes, Poly(2-oxazolines) & Dextrans

Poly-Amino Acids

Poly-Arg, Poly-Glu, Poly-Lys, Poly-Orn, Poly-Sar

PEGylation

Branched PEGylating reagents, (amino-)PEG-acids, PEG-amines & hydrazides & guanidines, reagents for Click-conjugation, Biotin-PEG-reagents, PEG-thiols, PEG-maleimides, other PEGylating reagents

Biotinylation Reagents

Carbohydrates

Galactose, Glucose, Maltose, Mannose, Xylose and others

Drug Metabolites

Peptides

Substrates & Inhibitors

E.g. protein kinase inhibitors, substrates for fusion (Halo/ Snap/Clip)-tagged proteins

Natural Products

Dyes and Fluorescent Labels

E.g. ICG, AMC, DAPI

Maillard & Amadori Reaction Products

Large portfolio of derivatives useful as standards for food, pharma and cosmetics industry

Vitamins

Custom Synthesis

Your project requires a compound not listed in our portfolio? Get in contact and inquire about our custom synthesis capabilities.

Our experienced scientists are excited to accept your synthetic challenge! In such cases, your request undergoes the following stages:

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Step-by-Step Analysis

• Customer’s demands

Process Evaluation

Detailed literature review

• Synthetic possibilities

Our Service Promise

Strategy Development

Protocol development

• Method development and validation

• Customized synthesis

Quality Consistency

Identity confirmation

• Purity verification

All our services are based on high standards, transparency & documentation, trust, honesty & confidentiality, as well as the required know-how.

High Standards

• Values: sustainability & responsibility

• State-of-the-art equipment & latest technologies

High quality standards

• Qualified suppliers & regular audits

Trust, Honesty & Confidentiality

Intergenerational business valuing partnerships

• Meeting the customer‘s expectations

Integrity towards our customers

Transparency & Documentation

• Talk to our specialists – customer care

• Certificates of analysis & impurity profiling

Analytical and process reports

Our Know-How

One-step reactions & complex multi-step synthesis

• Scalability from mg to kg quantities

Route scouting

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1. Photochemistry in Peptide Synthesis and Bioconjugation

The term photochemistry describes a group of chemical transformations initiated by irradiation with light. Photochemical reactions usually occur at room temperature and normal pressure, and mostly do not require additional reagents or catalysts, with the notable exception of some cases where the presence of a photosensitizer is necessary. Therefore, photochemical transformations are usually orthogonal to classical chemical transformations, a characteristic that renders them a valuable tool for chemists.

Consequently, the applications of photochemistry are numerous. Orthogonality is a trait often sought for in protecting groups and linkers, as it allows for their selective cleavage. Furthermore, photocleavage is a convenient method for selective removal of auxiliaries after their function has been served. The typical moiety that is incorporated into protecting groups, linkers and auxiliaries to facilitate light-induced cleavage is an o-nitrobenzyl group, which undergoes a Norrish-type II reaction upon UV-irradiation.

Another common application of photochemistry is the labeling or crosslinking of biomolecules in vitro and in vivo. The latter is of particular interest as a photochemical reaction is one of the few chemical transformations that can be selectively initiated in living cells.

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Do you require different photoactivated compounds?

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Inquire with our Custom Synthesis service and download our booklet.

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2. Diazirine Amino Acids for Photo-Crosslinkage in Living Cells

Iris Biotech introduces a comprehensive set of photo-crosslinking amino acids bearing the diazirine moiety. Irradiation of diazirines with UV light (ca. 350 nm – 360 nm) yields a highly reactive carbene species that can undergo insertions into C-C, CH, O-H and X-H (X = heteroatom) bonds of neighboring molecules to irreversibly form a covalent bond. The diazirine moiety is the smallest of all photophores, so introduction of a diazirine-bearing amino acid into a peptide or protein usually does not impair its biological activity. Further advantages of diazirine crosslinkers are their stability at room temperature, as well as their relative stability to nucleophiles, and to both acidic and basic conditions.

Applications:

• Probing protein–protein and protein- peptide interactions by photo-crosslinking (e.g. site-specific incorporation of Photo-Lys into glutathione S-transferase allowing covalent crosslinking of the two subunits of the dimeric protein in E. coli)

• Probing protein- peptide interactions in order to identify cellular targets of peptides of interest

Studying protein−drug interactions and identifying new drug targets By comparing results obtained from different proteomic setups (e.g. live cells and cell lysates), more putative targets can be identified.

These amino acids are available in Fmoc- as well as Boc-protected versions for their incorporation into synthetic peptides via standard coupling methods. Furthermore, the unprotected diazirine amino acids are also available for incorporation into expressed peptides and proteins by utilizing the appropriate aminoacyl-tRNA synthetase/tRNA pair. A combination of synthetic and recombinant approaches utilizing NCL has been demonstrated as well. Unnatural amino acids are frequently toxic to cells; however, these photo-amino acids are functional and nontoxic, which allows them to be a premium tool for studying mechanisms and interactions in living cells.

Fig. 1: Diazirine amino acids and their natural counterparts.

Photo-Phe

Fig. 2: Use of photo-phenylalanine for the identification of angiotensin-II-receptor binding sites; 125I is used as a radiotracer.

Photochemistry

HAA3100

H-L-Photo-Leucine*HCl

(S)-2-amino-3-(3-methyl-3H-diazirin-3-yl)propanoic acid hydrochloride

CAS-No. 851960-91-3

Formula C 5H9 N3 O 2*HCl

Mol. weight 143,14*36,45 g/mol

BAA3070

Boc-L-Photo-Leucine*DCHA

(S)-2-(tert-butoxycarbonylamino)-3-(3-methyl-3H-diazirin-3-yl)propanoic acid dicyclohexylamine

CAS-No. 1000770-97-7 net

Formula C10 H17N3 O4*C12H23 N

Mol. weight 243,26*181,34 g/mol

FAA4590 Fmoc-L-Photo-Leucine

(S)-2-(((9H-fluoren-9-yl)methoxy)carbonylamino)-3-(3-methyl-3H-diazirin-3-yl)propanoic acid

CAS-No. 1360651-24-6

Formula C 20 H19 N3 O4

Mol. weight 365,38 g/mol

HAA3110 H-L-Photo-Lysine*HCl

(S)-2-amino-6-((2-(3-methyl-3H-diazirin-3-yl)ethoxy) carbonylamino)hexanoic acid hydrochloride

CAS-No. 1253643-88-7

Formula C11H20 N4O4*HCl

Mol. weight 272,30*36,45 g/mol

FAA4600 Fmoc-L-Photo-Lysine

(S)-2-(((9H-fluoren-9-yl)methoxy)carbonylamino)-6((2-(3-methyl-3H-diazirin-3-yl)ethoxy)carbonylamino) hexanoic acid

CAS-No. 2250437-42-2

Formula C 26 H30 N4O 6

Mol. weight 494,54 g/mol

BAA3080 Boc-L-Photo-Lysine

(S)-2-(tert-butoxycarbonylamino)-6-((2-(3-methyl-3Hdiazirin-3-yl)ethoxy)carbonylamino)hexanoic acid

CAS-No. 1330088-06-6

Formula C16 H28 N4O 6

Mol. weight 372,42 g/mol

HAA3120

H-L-Photo-Methionine*HCl (S)-2-amino-4-(3-methyl-3H-diazirin-3-yl)butanoic acid hydrochloride

CAS-No. 851960-68-4 net

Formula C 6 H11N3 O 2*HCl

Mol. weight 157,17*36,45 g/mol

BAA3090

Boc-L-Photo-Methionine*DCHA

(S)-2-(tert-butoxycarbonylamino)-4-(3-methyl-3H-diazirin-3-yl)butanoic acid dicyclohexylamine

CAS-No. 1002754-75-7 net

Formula C11H19 N3 O4*C12H23 N

Mol. weight 257,29*181,34 g/mol

FAA4610

Fmoc-L-Photo-Methionine (S)-2-(((9H-fluoren-9-yl)methoxy)carbonylamino)-4-(3-methyl-3H-diazirin-3-yl)butanoic acid

CAS-No. 945859-89-2

Formula C 21H21N3 O4

Mol. weight 379,41 g/mol

HAA3490 H-L-Photo-Phe-OH

4-(trifluoromethyldiazirin)-L-phenylalanine

CAS-No. 92367-16-3

Formula C11H10 F 3 N3 O 2

Mol. weight 273,21 g/mol

BAA1530 Boc-L-Photo-Phe-OH

N-alpha-(t-Butyloxycarbonyl)-4-(trifluoromethyldiazirin)-L-phenylalanine

CAS-No. 92367-17-4

Formula C16 H18 F 3 N3 O4

Mol. weight 373,33 g/mol

FAA5690 Fmoc-L-Photo-Phe-OH

N-alpha-(9-Fluorenylmethyloxycarbonyl)-4-(trifluoromethyldiazirin)-L-phenylalanine

CAS-No. 133342-64-0

Formula C 26 H20 F 3 N3 O4

Mol. weight 495,45

HAA3130 H-L-Photo-Proline*HCl

(S)-1,2,5-triazaspiro[2.4]hept-1-ene-6-carboxylic acid

CAS-No. 1675206-55-9

Formula C 5H7N3 O 2*HCl

Mol. weight 141,13*36,45 g/mol

BAA3100

Boc-L-Photo-Proline

(S)-5-(tert-butoxycarbonyl)-1,2,5-triazaspiro[2.4]hept-1ene-6-carboxylic acid

CAS-No. 1266778-55-5

Formula C10 H15N3 O4

Mol. weight 241,24 g/mol

FAA4620

Fmoc-L-Photo-Proline

(S)-5-(((9H-fluoren-9-yl)methoxy)carbonyl)-1,2,5-triazaspiro[2.4]hept-1-ene-6-carboxylic acid

CAS-No. 1266778-58-8

Formula C 20 H17N3 O4 Mol. weight 363,37 g/mol

References:

→ Protein-polymer conjugation via ligand affinity and photoactivation of glutathione S-transferase; E. W. Lin, N. Boehnke, H. D. Maynard; Bioconjug Chem 2014; 25: 1902-9. https://doi.org/10.1021/bc500380r

→ Cell-based proteome profiling of potential dasatinib targets by use of affinity-based probes; H. Shi, C. J. Zhang, G. Y. Chen, S. Q. Yao; J Am Chem Soc 2012; 134: 3001-14. https://doi.org/10.1021/ja208518u

→ Probing protein-protein interactions with a genetically encoded photo-crosslinking amino acid; H. W. Ai, W. Shen, A. Sagi, P. R. Chen, P. G. Schultz; Chembiochem 2011; 12: 1854-7. https://doi.org/10.1002/cbic.201100194

→ Proteome profiling reveals potential cellular targets of staurosporine using a clickable cell-permeable probe; H. Shi, X. Cheng, S. K. Sze, S. Q. Yao; Chem Commun (Camb) 2011; 47: 11306-8. https://doi.org/10.1039/c1cc14824a

→ Aliphatic diazirines as photoaffinity probes for proteins: recent developments; J. Das; Chem Rev 2011; 111: 4405-17. https://doi.org/10.1021/cr1002722

→ Photo-crosslinking of proteins in intact cells reveals a dimeric structure of cyclooxygenase-2 and an inhibitor-sensitive oligomeric structure of microsomal prostaglandin E2 synthase-1; P. O. Hetu, M. Ouellet, J. P. Falgueyret, C. Ramachandran, J. Robichaud, R. Zamboni, D. Riendeau; Arch. Biochem. Biophys. 2008; 477: 155-62. https://doi.org/10.1016/j.abb.2008.04.038

→ Covalent capture of phospho-dependent protein oligomerization by site-specific incorporation of a diazirine photo-cross-linker; M. Vila-Perello, M. R. Pratt, F. Tulin, T. W. Muir; J Am Chem Soc 2007; 129: 8068-9. https://doi.org/10.1021/ja072013j

→ Photo-leucine incorporation reveals the target of a cyclodepsipeptide inhibitor of cotranslational translocation; A. L. MacKinnon, J. L. Garrison, R. S. Hegde, J. Taunton; J Am Chem Soc 2007; 129: 14560-1. https://doi.org/10.1021/ja076250y

→ Synthesis of photoactive analogues of a cystine knot trypsin inhibitor protein; T. Durek, J. Zhang, C. He, S. B. Kent; Org Lett 2007; 9: 5497-500. https://doi.org/10.1021/ol702461z

→ Photo-leucine and photo-methionine allow identification of protein-protein interactions in living cells; M. Suchanek, A. Radzikowska, C. Thiele; Nat Methods 2005; 2: 261-7. https://doi.org/10.1038/nmeth752

3. Photo-Crosslinkers for Various Applications

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Many photo-crosslinkers for photoaffinity labeling rely on benzophenone as the crosslinking agent. However, these types of crosslinkers usually show low crosslinking yields and require relatively long irradiation times due to slow reaction rates, which may lead to non-specific labeling. Moreover, the irradiation conditions for benzophenones have been shown to lead to cell damage and cell death.

Conversely, diazirine- and perfluorophenyl-based crosslinkers generate a reactive species (carbene and nitrene, respectively) upon relatively short irradiation with UV light. Consequently, diazirine- and perfluorophenyl-crosslinkers are commonly used in molecular biology and biochemistry.

3.1. Photo-Crosslinkers

Diazirine-bearing crosslinkers are activated at wavelengths that cause little to no damage to cells. In addition to this, trifluoromethylaryl diazirines show a fast initial reaction rate and rapid termination of reaction. Consequently, they exhibit a highly ligand-dependent reactivity, which renders them ideal probes for ligand binding to low-affinity targets, e.g. for probing carbohydrate-lectin interactions. Moreover, the small size of the diazirine group minimizes the risk of impairing or altering the biological activity of a ligand. Our diazirine crosslinkers are functionalized to react with either carboxyl- or aminereactive ligands, respectively.

Another application of diazirine photo crosslinkers is the surface immobilization of molecules of interest. The linker is bound to a surface through its carboxyl or amino functionality, leaving the diazirine group free to react with any type of molecule. Since this reaction takes place irrespective of available functional groups, it is not necessary to chemically modify molecules of interest prior to immobilization. This virtually ensures that molecules are immobilized without altering their binding properties. By using this approach, it is possible to easily create microarrays of whole libraries of small molecules for rapid screening. back to

Fig. 3: Three types of photophores: Perfluorophenyl azides, diazirines and benzophenones.

Photochemistry

Aryl/ alkyl-spacer

R2

R2

Aryl- / alkyl-spacer

Aryl/ alkyl-spacer

Aryl- / alkyl-spacer Probe

UV

R1 = COOH or CH2NH2

Photo-Pentanoic acid

CAS-No. 25055-86-1

Formula C 5H 8 N2O 2

Mol. weight 128,13 g/mol

OH O NN

RL-2900

Photo-Hexanoic acid

CAS-No. 16297-97-5

Formula C 6 H10 N2O 2

Mol. weight 142,16 g/mol

OH O N N

Fig. 4: Immobilization of molecules of interest on a surface (e.g. a glass slide) using a diazirine crosslinker.

Product details

RL-3410

Photo-Click-Heptanoic acid

2-(3-(but-3-ynyl)-3H-diazirin-3-yl)acetic acid

CAS-No. 2049109-24-0

Formula C 7H 8 N2O 2 Mol. weight 152,15 g/mol

RL-3740

Photo-Palmitic acid

9-(3-hexyl-3H-diazirin-3-yl)nonanoic acid

CAS-No. 2100292-41-7

Formula C16 H30 N2O 2 Mol. weight 282,43 g/mol

RL-3720

Photo-Click-Palmitic acid

11-(3-(but-3-yn-1-yl)-3H-diazirin-3-yl)undecanoic acid

Formula C16 H26 N2O 2 Mol. weight 278,40 g/mol

RL-3750

Photo-Stearic acid

11-(3-hexyl-3H-diazirin-3-yl)undecanoic acid

CAS-No. 1704120-02-4

Formula C18 H34N2O 2 Mol. weight 310,48 g/mol

RL-3760

Photo-Click-Stearic acid

13-(3-(but-3-yn-1-yl)-3H-diazirin-3-yl)tridecanoic acid

Formula C18 H30 N2O 2 Mol. weight 306,45 g/mol

RL-2920

Photo-Benzoic acid

4-[3-(Trifluoromethyl)-3H-diazirin-3-yl]benzoic acid

CAS-No. 85559-46-2

Formula C9 H 5F 3 N2O 2

RL-2910 Photo-Butylamine

2-(3-methyl-3H-diazirin-3-yl)ethan-1-amine hydrochloride NH2

CAS-No. 25055-95-2

Formula C 4H9 N3*HCl

Mol. weight 99,13*36,45 g/mol

RL-2930 Photo-Benzylamine*HCl

4-[3-(Trifluoromethyl)-3H-diazirin-3-yl]benzylamine hydrochloride

CAS-No. 1258874-29-1

Formula C9 H 8 N3 F 3*HCl

Mol. weight 215,18*36,45 g/mol

References:

Reviews:

→ Hide and seek: Identification and confirmation of small molecule protein targets; A. Ursu, H. Waldmann; Bioorg Med Chem Lett 2015; 25: 3079-86. https://doi.org/10.1016/j.bmcl.2015.06.023

→ Development and leading-edge application of innovative photoaffinity labeling; Y. Hatanaka; Chem Pharm Bull (Tokyo) 2015; 63: 1-12.

https://doi.org/10.1248/cpb.c14-00645

→ Diazirine based photoaffinity labeling; L. Dubinsky, B. P. Krom, M. M. Meijler; Bioorg Med Chem 2012; 20: 554-70.

https://doi.org/10.1016/j.bmc.2011.06.066

→ Aliphatic diazirines as photoaffinity probes for proteins: recent developments; J. Das; Chem Rev 2011; 111: 4405-17.

https://doi.org/10.1021/cr1002722

→ Recent Progress in Diazirine-Based Photoaffinity Labeling; M. Hashimoto, Y. Hatanaka; Eur. J. Org. Chem. 2008; 2008: 2513-2523 .

https://doi.org/10.1002/ejoc.200701069

→ Endeavors to make the photophore, diazirine easy to use; Y. Sadakane; Yakugaku Zasshi 2007; 127: 1693-9.

https://doi.org/10.1248/yakushi.127.1693

Photoaffinity labeling:

→ Comparison of the reactivity of carbohydrate photoaffinity probes with different photoreactive groups; K. Sakurai, S. Ozawa, R. Yamada, T. Yasui, S. Mizuno; Chembiochem 2014; 15: 1399-403. https://doi.org/10.1002/cbic.201402051

→ Identification of a substrate-binding site in a peroxisomal beta-oxidation enzyme by photoaffinity labeling with a novel palmitoyl derivative; Y. Kashiwayama, T. Tomohiro, K. Narita, M. Suzumura, T. Glumoff, J. K. Hiltunen, P. P. Van Veldhoven, Y. Hatanaka, T. Imanaka; J Biol Chem 2010; 285: 26315-25.

https://doi.org/10.1074/jbc.M110.104547

→ Developing photoactive affinity probes for proteomic profiling: hydroxamate-based probes for metalloproteases; E. W. Chan, S. Chattopadhaya, R. C. Panicker, X. Huang, S. Q. Yao; J Am Chem Soc 2004; 126: 14435-46. https://doi.org/10.1021/ja047044i

→ Insecticidal and neural activities of candidate photoaffinity probes for neonicotinoid binding sites; K. Matsuda, M. Ihara, K. Nishimura, D. B. Sattelle, K. Komai; Biosci Biotechnol Biochem 2001; 65: 1534-41.

https://doi.org/10.1271/bbb.65.1534

→ Synthesis of Photoactive α-Mannosides and Mannosyl Peptides and Their Evaluation for Lectin Labeling; M. Wiegand, T. K. Lindhorst; Eur. J. Org. Chem. 2006; 2006: 4841-4851. https://doi.org/10.1002/ejoc.200600449

Photoaffinity microarray:

→ A study on photolinkers used for biomolecule attachment to polymer surfaces; D. M. Dankbar, G. Gauglitz; Anal Bioanal Chem 2006; 386: 1967-74. https://doi.org/10.1007/s00216-006-0871-x

→ SPR imaging of photo-cross-linked small-molecule arrays on gold; N. Kanoh, M. Kyo, K. Inamori, A. Ando, A. Asami, A. Nakao, H. Osada; Anal Chem 2006; 78 : 2226-30. https://doi.org/10.1021/ac051777j

→ Grafting Organic and Biomolecules on H-Terminated Porous Silicon from a Diazirine; S. Wei, J. Wang, D.-J. Guo, Y.-Q. Chen, S.-J. Xiao; Chem. Lett. 2006; 35: 1172-1173. https://doi.org/10.1246/cl.2006.1172

→ Immobilization of natural products on glass slides by using a photoaffinity reaction and the detection of protein-small-molecule interactions; N. Kanoh, S. Kumashiro, S. Simizu, Y. Kondoh, S. Hatakeyama, H. Tashiro, H. Osada; Angew. Chem. Int. Ed. 2003; 42: 5584-7. https://doi.org/10.1002/anie.200352164

3.2. Tetrafluorophenyl-Azide-based Photo-Crosslinkers

Tetrafluorophenyl-azides follow a principle similar to diazirines. Upon irradiation with UV light (ca. 260 nm), a highly stabilized nitrene is formed. Nitrenes are the nitrogen analogs of carbenes (isoelectronic) and react in a comparable fashion. In terms of crosslinking yield and duration of irradiation, they compare favorably to benzophenones. Moreover, the azido group can also undergo classical copper-catalyzed azide-alkyne cycloadditions. Tetrafluorophenyl-azido crosslinkers are also available with a short PEG-spacer for increased solubility (PEG5000), and as Biotin-TEG-ATFBA (PEG2065) for applications such as surface functionalization with biotin, or the biotinylation of biomacromolecules.

Fig. 6: Applications of Tetrafluorophenyl-azides.

Photochemistry

RL-2035 ATFB

4-Azido-2,3,5,6-tetrafluorobenzoic acid

CAS-No. 122590-77-6

Formula C 7HF4N3 O 2

Mol. weight 235,1 g/mol

RL-2045 ATFB-NHS

N-Succinimidyl 4-azido-2,3,5,6-tetrafluorobenzoate

CAS-No. 126695-58-7

Formula C11H4F4N4O4

Mol. weight 332,17 g/mol

PEG2065 Biotin-TEG-ATFBA

Biotin-triethylenglycol-( p-azido-tetrafluorobenzamide)

CAS-No. 1264662-85-2

Formula C 27H37F4N7 O 6 S

Mol. weight 663,68 g/mol

PEG5000 N3 -TFBA-O2Oc

{2-[2-(4-Azido-2,3,5,6-tetrafluorobenzoyl-amino)ethoxy] ethoxy}acetic acid

CAS-No. 1993119-45-1

Formula C13 H12F4N4O 5

Mol. weight 380,25 g/mol

References:

→ Tri- and Tetravalent Photoactivable Cross-Linking Agents; A. Welle, F. Billard, J. Marchand-Brynaert; Synthesis 2012; 44: 2249-2254. https://doi.org/doi:10.1055/s-0031-1290444

→ Perfluorophenyl azides: new applications in surface functionalization and nanomaterial synthesis; L. H. Liu, M. Yan; Acc Chem Res 2010; 43: 1434-43. https://doi.org/10.1021/ar100066t

→ Photo-click immobilization of carbohydrates on polymeric surfaces--a quick method to functionalize surfaces for biomolecular recognition studies; O. Norberg, L. Deng, M. Yan, O. Ramstrom; Bioconjug Chem 2009; 20: 2364-70. https://doi.org/10.1021/bc9003519

→ Photoreactive insulin derivatives for the detection of the doubly labeled insulin receptor; J. Kleinjung, M. Fabry; Peptides 2000; 21: 401-6. https://doi.org/10.1016/S0196-9781(00)00164-9

→ Recent Trends in the Evaluation of Photochemical Insertion Characteristics of Heterobifunctional Perfluoroaryl Azide Chelating Agents: Biochemical Implications in Nuclear Medicine; R. S. Pandurangi, S. R. Karra, R. R. Kuntz, W. A. Volkert; Photochem. Photobiol. 1997; 65: 208-221. https://doi.org/10.1111/j.1751-1097.1997.tb08547.x

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4. Photoactivated Self-Cleaving Linkers and Protecting Groups via Trimethyl Lock

Iris Biotech introduces a series of self-immolative compounds that find application as protecting groups, linkers, or amino acid derivatives (Spr = stimulus-responsive peptide bond cleaving residue). The selfcleavage is induced by irradiation with UV light (ca. 350 – 365 nm) that leads to the unmasking of a hydroxyl group of a 2-alkyl-3,5-dimethyl phenol moiety. The photocleavable group is either o-nitrobenzyl or o-nitroveratryl, which can be cleaved at wavelengths > 350 nm. Since wavelengths above 350 nm tend to be unproblematic for biomacromolecules, this technique is especially interesting for cell-based systems.

The liberated OH-group serves as a nucleophile that intramolecularly cleaves ester (or amide) bonds at neutral pH and room temperature by cyclization via a six-membered transition state. This reaction is greatly accelerated since the sterically unfavorable interaction between the methyl group at the 3-position of the phenol core and the two geminal CH 3 -groups on the alkyl chain (in β-position to the ester or amide carbonyl group) favor conformations that bring the phenolic OH-group and the neighboring carbonyl function into closer vicinity. This phenomenon is termed the gem-dialkyl effect, for which a theory was first proposed by Thorpe and Ingold in 1915 (“Thorpe-Ingold effect”). An alternative explanation for this effect was posited by Bruice and Pandit in 1960 (“reactive rotamer effect”).

The applications for this approach are numerous. Incorporation of a Spr-residue into a peptide sequence enables the photoactivated self-cleavage of the peptide at the position of said residue. This technique allows for the intracellular removal of e.g. cell penetrating peptides or localization sequences from a bioactive molecule. The Spr-residue can also be used as a photolabile linker in order to reversibly connect a moiety such as biotin to a molecule of interest. Finally, while o-nitroveratryl itself is a valuable protecting group for sulfhydryl groups, the combination of o-Nv with a trimethyl lock moiety also allows for its use as a protecting group for hydroxyl and amino functions.

Fig. 7: Principle of photoactivated trimethyl lock; PG = o-nitrobenzyl or o-nitroveratryl; X = O or NH; R1–XH = target molecule; R2 = H or NH-alkyl.

FAA7190 Fmoc-Spr(oNB)-OH

N-alpha-(9-Fluorenylmethyloxycarbonyl)-beta,beta-dimethyl-(2,4-dimethyl-6-(2-nitrobenzyloxy)phenyl) alanine (rac.)

CAS-No. 1032400-98-8

Formula C 35H34N2O 7

Mol. weight 594,66 g/mol

FAA7200 Fmoc-Spr(o Nv)-OH

N-alpha-(9-Fluorenylmethyloxycarbonyl)-beta,beta-dimethyl-(2-methyl-6-(2-nitroveratryl)phenyl)alanine (rac.)

CAS-No. 1228829-20-6

Formula C 36 H36 N2O 9

Mol. weight 640,68 g/mol

RL-2970 Photo-Trimethyl-Lock

3-(2-Nitroveratryl-4,6-dimethylphenyl)-3-methylbutyric acid

CAS-No. 2095134-25-9

Formula C 22H27NO 7 Mol. weight 417,45 g/mol

References:

→ Invention of stimulus-responsive peptide-bond-cleaving residue (Spr) and its application to chemical biology tools; A. Shigenaga, J. Yamamoto, T. Kohiki, T. Inokuma, A. Otaka; J. Pept. Sci. 2017; 23: 505-513.

https://doi.org/10.1002/psc.2961

→ Syntheses and kinetic studies of cyclisation-based self-immolative spacers; S. Huvelle, A. Alouane, T. Le Saux, L. Jullien, F. Schmidt; Org Biomol Chem 2017; 15: 3435-3443. https://doi.org/10.1039/c7ob00121e

→ Photo-triggered fluorescent labelling of recombinant proteins in live cells; D. Jung, K. Sato, K. Min, A. Shigenaga, J. Jung, A. Otaka, Y. Kwon; Chem Commun (Camb) 2015; 51: 9670-3.

https://doi.org/10.1039/c5cc01067e

→ Trimethyl lock: A trigger for molecular release in chemistry, biology, and pharmacology; M. N. Levine, R. T. Raines; Chem. Sci. 2012; 3: 2412-2420. https://doi.org/10.1039/C2SC20536J

→ Design and synthesis of caged ceramide: UV-responsive ceramide releasing system based on UV-induced amide bond cleavage followed by O–N acyl transfer; A. Shigenaga, H. Hirakawa, J. Yamamoto, K. Ogura, M. Denda, K. Yamaguchi, D. Tsuji, K. Itoh, A. Otaka; Tetrahedron 2011; 67: 3984-3990. https://doi.org/10.1016/j.tet.2011.04.048

→ Development and photo-responsive peptide bond cleavage reaction of two-photon near-infrared excitationresponsive peptide; A. Shigenaga, J. Yamamoto, Y. Sumikawa, T. Furuta, A. Otaka; Tetrahedron Lett. 2010; 51: 2868-2871. https://doi.org/10.1016/j.tetlet.2010.03.079

→ gem-disubstituent effect: theoretical basis and synthetic applications; M. E. Jung, G. Piizzi; Chem Rev 2005; 105: 1735-66. https://doi.org/10.1021/cr940337h

→ The Effect of Geminal Substitution Ring Size and Rotamer Distribution on the Intramolecular Nucleophilic Catalysis of the Hydrolysis of Monophenyl Esters of Dibasic Acids and the Solvolysis of the Intermediate Anhydrides; T. C. Bruice, U. K. Pandit; J. Am. Chem. Soc. 1960; 82: 5858-5865. https://doi.org/10.1021/ja01507a023

→ CXIX.—The formation and stability of spiro-compounds. Part I. spiro-Compounds from cyclohexane; R. M. Beesley, C. K. Ingold, J. F. Thorpe; J. Chem. Soc., Trans. 1915; 107: 1080-1106. https://doi.org/10.1039/ct9150701080

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5. Furfuryl-Alanine for Side Chain Modification and Bioconjugation

2-Furyl-alanine can be incorporated into peptides via SPPS or by using enzymatic approaches. UV-irradiation in the presence of oxygen and a photosensitizer converts furyl-alanine to an intermediate that selectively reacts with certain nucleophiles. This property can be employed for site-specific labeling of peptides and proteins.

Labeling with different tags and reporter groups is a pivotal technique for the elucidation of peptide and protein function.

A novel and innovative approach is the site-specific labeling using the unnatural amino acid 2-furylalanine. UV-irradiation in the presence of oxygen and a photosensitizer converts furyl-alanine to an unsaturated dicarbonyl compound. This intermediate selectively reacts with certain nucleophiles such as hydrazine derivatives of dyes or fluorescent labels. This reaction can be used for the site-specific labeling of peptides and proteins and can be carried out in aqueous solution.

Iris Biotech offers Fmoc-L-Ala(2-Furyl)-OH suitable for SPPS, as well as H-L-Ala(2-Furyl)-OH which can be incorporated into proteins using the amber suppression methodology.

HAA2930 H-L-Ala(2-Furyl)-OH

3-(2-Furyl)-L-alanine

CAS-No. 127682-08-0

Mol. weight 155,15 g/mol

Fig. 8: Site-specific labeling of peptides and proteins using 2-Furyl-alanine.

FAA4250 Fmoc-L-Ala(2-Furyl)-OH

N-alpha-(9-Fluorenylmethyloxycarbonyl)-3-(2-furyl)-L-alanine

CAS-No. 159611-02-6

Formula C 22H19 NO 5

Mol. weight 377,39 g/mol

References:

→ Novel furan-oxidation based site-specific conjugation methodology for peptide labeling and antibody drug conjugates; presented at the 6th World ADC meeting October 19 th-22nd 2015, San Diego, CA; An Van Den Bulcke, Eirini Antonatou, Willem Vannecke, Kurt Hoogewijs, Annemieke Madder.

→ Exploiting furan‘s versatile reactivity in reversible and irreversible orthogonal peptide labeling; K. Hoogewijs, D. Buyst, J. M. Winne, J. C. Martins, A. Madder; Chem Commun (Camb) 2013; 49: 2927-9.

https://doi.org/10.1039/c3cc40588e

→ Sequence specific DNA cross-linking triggered by visible light; M. Op de Beeck, A. Madder; J Am Chem Soc 2012; 134: 10737-40. https://doi.org/10.1021/ja301901p

→ Unprecedented C-selective interstrand cross-linking through in situ oxidation of furan-modified oligodeoxynucleotides; M. Op de Beeck, A. Madder; J Am Chem Soc 2011; 133: 796-807. https://doi.org/10.1021/ja1048169

→ Furan-modified oligonucleotides for fast, high-yielding and site-selective DNA inter-strand cross-linking with non-modified complements; K. Stevens, A. Madder; Nucleic Acids Res 2009; 37: 1555-65. https://doi.org/10.1093/nar/gkn1077

→ From DNA cross-linking to peptide labeling: on the versatility of the furan-oxidation-conjugation strategy; A. Deceuninck, A. Madder; Chem Commun (Camb) 2009; 340-2. https://doi.org/10.1039/b817447d

→ Bioorthogonal chemistry: fishing for selectivity in a sea of functionality; E. M. Sletten,C. R. Bertozzi; Angew. Chem. Int. Ed. 2009; 48: 6974-98.

https://doi.org/10.1002/anie.200900942

→ Structural basis of furan-amino acid recognition by a polyspecific aminoacyl-tRNA-synthetase and its genetic encoding in human cells; M. J. Schmidt, A. Weber, M. Pott, W. Welte, D. Summerer; Chembiochem 2014; 15: 1755-60.

https://doi.org/10.1002/cbic.201402006

→ Red-light-controlled protein-RNA crosslinking with a genetically encoded furan; M. J. Schmidt, D. Summerer; Angew. Chem. Int. Ed. 2013; 52: 4690-3.

https://doi.org/10.1002/anie.201300754

. 6. Photocaged Amino Acid Building Blocks

Photocaged amino acids offer a mild and selective way to deprotect reactive side-chain functional groups such as amines or thiols, thus adding another level of orthogonality to the peptide chemist’s toolbox. Photolabile protecting groups (PGs) are frequently based on the ortho-nitrobenzyl moiety or variations thereof. Upon irradiation with UV light, such nitrobenzyl-based PGs undergo a Norrish type II reaction, liberating the protected functional group.

A common modification to the ortho-nitrobenzyl motif is the addition of two methoxy groups to the phenyl ring. The resulting ortho-nitroveratryl (oNv) group is compatible with SPPS protocols and can be cleaved by irradiation with UV light (350 nm) under ambient conditions.

One possible application of the oNv PG is the regiospecific formation of disulfide bonds. When combined with S-pyridinesulfenyl activation by employing Fmoc-L-Cys(Npys)-OH (FAA1975), the Fmoc-L-Cys(oNv)-OH (FAA3970) building block allows for a rapid in situ SS-bond formation. The versatility of this approach was demonstrated by applying it in the synthesis of a number of model peptides such as oxytocin, alpha-conotoxin ImI, and human insulin.

Fig. 9: Selective deprotection of 2-nitroveratryl in the presence of other cysteine protecting groups, and subsequent disulfide bond formation with an Npys-functionalized cysteine.

The oNv group can also be employed to protect the side chain of lysine if it is used as the corresponding oxycarbonyl derivative, abbreviated Nvoc (o-nitroveratryloxycarbonyl). The corresponding building block Fmoc-L-Lys(Nvoc)-OH (FAA7230) is available at Iris Biotech. Additionally, o-nitroveratryl may be used as a solubilizing tag on the alpha-amino group of glycine for internal solubilization, which can be cleaved by photolysis (350 nm) under ambient conditions.

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Photochemistry

Another variation of the o-nitrobenzyl motif is the nitrodibenzofuran (NDBF) group that has an additional benzofuran ring system fused to the existing phenyl ring. NDBF is a one- and two-photon sensitive protecting group for cysteine. Analogous to the oNv group, NDBF can also be used to protect the epsilon amino group of lysine in the form of the corresponding oxycarbonyl derivative NDBFOC. Both Fmoc-LCys(NDBF)-OH (FAA8420) and Fmoc-L-Lys(NDBFOC)-OH (FAA8425) are available at Iris Biotech as readyto-use building blocks for peptide synthesis. Selective deprotection by photolysis is achieved either by irradiation with UV light (365 nm), or by two-photon excitation with near-infrared (800 nm) light. The latter aspect renders this derivative particularly useful for biological applications in live cells or intact organisms due to the increased penetration depth of NIR light, as well as its biocompatibility. In addition, this cage exhibits a faster UV photolysis rate relative to simple nitroveratryl derivatives. An example showed that NDBF is photolyzed 16–160 times more efficiently than other nitrobenzyl PPGs. Further variations include the dimethoxynitrobenzyl group, as well as the methyl-o-nitropiperonyl group, which are used to protect selenocysteine and cysteine, respectively.

FAA3970 Fmoc-L-Cys(o Nv)-OH

N-alpha-(9-Fluorenylmethyloxycarbonyl)-S-(2-nitroveratryl)-L-cysteine

CAS-No. 214633-71-3

Formula C 27H26 N2O 8 S

Mol. weight 538,57 g/mol

FAA1975 Fmoc-L-Cys(Npys)-OH

N-alpha-(9-Fluorenylmethyloxycarbonyl)-S-(3-nitro-2-pyridylthio)-L-cysteine

CAS-No. 159700-51-3

Formula C 23 H19 N3 O 6 S 2

Mol. weight 497,54 g/mol

FAA7230 Fmoc-L-Lys(Nvoc)-OH

N-alpha-(9-Fluorenylmethyloxycarbonyl)-N-epsilon-(o-nitroveratryloxycarbonyl)-L-lysine

CAS-No. 150571-28-1

Formula C 31H33 N3 O 10

Mol. weight 607,61 g/mol

FAA8420 Fmoc-L-Cys(NDBF)-OH

N-alpha-(9-Fluorenylmethyloxycarbonyl)-S-(1-(3-nitro-dibenzofuran-2-yl)-ethyl)-L-cysteine

CAS-No. 1895883-28-9

Formula C 32H26 N2O 7S

Mol. weight 582,62 g/mol

FAA8425 Fmoc-L-Lys(NDBFOC)-OH

N-alpha-(9-Fluorenylmethyloxycarbonyl)-N-epsilon-(1-(3-nitro-dibenzofuran-2-yl)-ethoxycarbonyl)-L-lysine

Formula C 36 H33 N3 O 9

HAA9360 H-L-Sec(MDNPE)-OH

Se-(Methyl-o-nitropiperonyl)-selenocysteine

CAS-No. 2235373-47-2

Formula C12H14N2O 6 Se

Mol. weight 361,21 g/mol

HAA9255 H-L-Sec(DMNB)-OH*TFA

Dimethoxynitrobenzyl selenocysteine TFA salt

CAS-No. 1644398-13-9

Formula C12H16 N2O 6 Se*CF 3 COOH

Mol. weight 363,24*114,02 g/mol

HAA9320 H-L-Cys(DMNB)-OH

S-(4,5-dimethoxy-2-nitrobenzyl)-L-cysteine

CAS-No. 214633-68-8

Formula C12H16 N2O 6 S

Mol. weight 316,33 g/mol

HAA9270 H-L-Cys(MDNPE)-OH

1-[4‘,5‘-(methylenedioxy)-2‘-nitrophenyl]ethyl]-L-cysteine

CAS-No. 1551078-43-3

Formula C12H14N2O 6 S

Mol. weight 314,31 g/mol

FAA7945 Fmoc-L-Cys(MDNPE)-OH

N-(((9H-fluoren-9-yl)methoxy)carbonyl)-S-(1-(6-nitrobenzo[d][1,3]dioxol-5-yl)ethyl)-L-cysteine

Formula C 27H24N2O 8 S

Mol. weight 536,56 g/mol

Photochemistry

FAA8870 Fmoc-L-hCys(o Nv)-OH

N-alpha-(9-Fluorenylmethyloxycarbonyl)-S-(2-nitroveratryl)-L-homocysteine

Formula C 28 H28 N2O 8 S Mol. weight 552,60 g/mol

FAA7350 Fmoc-N(

o Nv)-Gly-OH

N-alpha (9-Fluorenylmethyloxycarbonyl)-N-alpha-(o-nitroveratryl)-glycine

CAS-No. 850859-64-2

Formula C 26 H24N2O 8

Mol. weight 492,48 g/mol

FAA7200 Fmoc-Spr(o Nv)-OH

N-alpha-(9-Fluorenylmethyloxycarbonyl)-beta,beta-dimethyl-(2-methyl-6-(2-nitroveratryl)phenyl)alanine (rac.)

CAS-No. 1228829-20-6

Formula C 36 H36 N2O 9

Mol. weight 640,68 g/mol

HAA9465 H-L-Sec(oNB)-OH*HCl

(R)-2-amino-3-((2-nitrobenzyl)selanyl)propanoic acid

CAS-No. 324582-23-2 net

Formula C10 H12N2O4 Se*HCl

Mol. weight 303,18*36,46 g/mol

HAA9475 H-L-Sec(NPE)-OH*HCl

(2R)-2-amino-3-((1-(2-nitrophenyl)ethyl)selanyl)propanoic acid

Formula C11H14N2O4 Se*HCl

Mol. weight 317,02*36,46 g/mol

References:

→ Total chemical synthesis of photoactivatable proteins for light-controlled manipulation of antigen–antibody interactions; S. Tang, Z. Wan, Y. Gao, J.-S. Zheng, J. Wang, Y.-Y. Si, X. Chen, H. Qi, L. Liu, W. Liu; Chem. Sci. 2016; 7: 1891-1895. https://doi.org/10.1039/C5SC03404C

→ 2-Nitroveratryl as a Photocleavable Thiol-Protecting Group for Directed Disulfide Bond Formation in the Chemical Synthesis of Insulin; J. A. Karas, D. B. Scanlon, B. E. Forbes, I. Vetter, R. J. Lewis, J. Gardiner, F. Separovic, J. D. Wade, M. A. Hossain; Chemistry 2014; 20: 9549-9552. https://doi.org/10.1002/chem.201403574

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→ Synthesis, stability and optimized photolytic cleavage of 4-methoxy-2-nitrobenzyl backbone-protected peptides; E. C. B. Johnson, S. B. H. Kent; Chem. Commun. 2006; 1557-1559 https://doi.org/10.1039/B600304D

→ A red shifted two-photon-only caging group for three-dimensional photorelease. Y. Becker, E. Unger, M. A. H. Fichte, D. A. Gacek, A. Dreuw, J. Wachtveitl, P. J. Walla, A. Heckel; Chem. Sci. 2018; 9: 2797-2902.

https://doi.org/10.1039/c7sc05182d

→ Nitrodibenzofuran: A One- and Two-Photon Sensitive Protecting Group That Is Superior to Brominated Hydroxycoumarin for Thiol Caging in Peptides. M. M. Mahmoodi, D. Abate-Pella, T. J. Pundsack, C. C. Palsuledesai, P. C. Goff, D. A. Blank, M. D. Distefano; J. Am. Chem. Soc. 2016; 138: 5848-5859.

https://doi.org/10.1021/jacs.5b11759

→ The nitrodibenzofuran chromophore: a new caging group for ulta-efficient photolysis in living cells.

A. Momotake, N. Lindegger, E. Niggli, R. J. Barsotti, G. C. R. Ellis-Davies; Nat. Methods 2006; 3: 35-40.

https://doi.org/10.1038/NMETH821

→ Methoxy-Substituted Nitrodibenzofuran-Based Protecting Group with an Improved Two-Photon Action Cross-Section for Thiol Protection in Solid Phase Peptide Synthesis. T. K. Bader, F. Xu, M. H. Hodny, D. A. Blank, M. D. Distefano; J. Org. Chem. 2020; 85: 1614-1625. https://doi.org/10.1021/acs.joc.9b02751.

→ Genetic Encoding of Photocaged Cysteine Allows Photoactivation of TEV Protease in Live Mammalian Cells; D. P. Nguyen, M. Mahesh, S. J. Elsässer, S. M. Hancock, C. Uttamapinant, J. W. Chin; J. Am. Chem. Soc. 2014; 136(6) : 2240-2243. https://doi.org/10.1021/ja412191m

→ Biosynthetic selenoproteins with genetically-encoded photocaged selenocysteines; R. Rakauskaité, G. Urbanavičiūtė, A. Rukšėnaitė, Z. Liutkevičiūtė, R. Juškėnas, V. Masevičiusab, S. Klimašauskas; Chem. Commun. 2015; 51: 8245-8248. https://doi.org/10.1039/C4CC07910H

Interested in Cysteines and disulfide bond formation?

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Discover our brochure about Cyclic Peptides.

Photochemistry

. 7. Photoswitches

Photoswitches are a subtype of molecular switch that undergo a reversible structural change upon irradiation with light to adopt a different configuration. Activation of a molecular switch by light offers several advantages. In addition to being traceless, light can be precisely controlled in its intensity (dosage control) and can be focused with sub-micron accuracy with a high temporal and spatial resolution. Consequently, photoswitches ideally lend themselves to the construction of light-responsive pharmaceutical compounds, a field that is described by the term photopharmacology.

The most common motif that is used in this approach is the azobenzene moiety, owing to the large geometrical change that results from adopting a different configuration. Azobenzene-based photoresponsive systems have also been successfully applied in several biological systems. The azobenzene moiety is synthetically accessible via various pathways (Mills reaction, oxidative/reductive coupling, azo coupling), and its photochromic properties can easily be finetuned. The thermodynamically more stable trans isomer adopts an extended planar configuration with a dipole moment close to zero, while the higher energy, metastable cis isomer is more polar and exhibits unfavorable steric interactions.

An additional benefit of the azobenzene motif is its high degree of reversibility. It can undergo approximately 100 switching cycles without detectable photodegradation or loss of responsiveness. However, although the trans isomer can be regenerated 100% by thermal relaxation, full conversion of one or the other isomer by irradiation with visible light is impossible owing to a substantial overlap of the absorption spectra of both isomers.

4-Phenylazo-L-phenylalanine hydrochloride

CAS-No. 2137036-84-9

Formula C15H15N3 O 2*HCl

Mol. weight 269,30*36,45 g/mol

Fig. 10: Interconversion between the trans and cis isomers of the diazobenzene motif in a photoswitchable peptide.

BAA1660

Boc-L-Phe(4-N=NPh)-OH

N-alpha-t-Butyloxycarbonyl-4-phenylazo-L-phenylalanine

CAS-No. 134816-33-4

Formula C 20 H23 N3 O4

Mol. weight 369,41 g/mol

FAA7010

Fmoc-L-Phe(4-N=NPh)-OH

N-alpha-(9-Fluorenylmethyloxycarbonyl)-4-phenylazo-L-phenylalanine

CAS-No. 210366-26-0

Formula C 30 H25N3 O4

Mol. weight 491,54 g/mol

FAA9180

Fmoc-AMPB-OH

4-(4-(9-Fluorenylmethoxycarbonylaminomethyl)-phenyldiazenyl)benzoic acid

CAS-No. 159790-79-1

Formula C 29 H23 N3 O4

Mol. weight 477,52 g/mol

References:

→ Photopharmacology and Photochemical Biology; A. Deiters, S. Kossatz, O. Vázquez; ChemPhotoChem 2021; 5: 1031-1032. https://doi.org/10.1002/cptc.202100216

→ Photoswitchable peptides for spatiotemporal control of biological functions; L. Albert, O. Vazquez; Chem Commun (Camb) 2019; 55: 10192-10213. https://doi.org/10.1039/c9cc03346g

→ In Vivo Photopharmacology; K. Hull, J. Morstein, D. Trauner; Chem Rev 2018; 118: 10710-10747. https://doi.org/10.1021/acs.chemrev.8b00037

→ Photopharmacological control of bipolar cells restores visual function in blind mice; L. Laprell, I. Tochitsky, K. Kaur, M. B. Manookin, M. Stein, D. M. Barber, C. Schon, S. Michalakis, M. Biel, R. H. Kramer, M. P. Sumser, D. Trauner, R. N. Van Gelder; J Clin Invest 2017; 127: 2598-2611. https://doi.org/10.1172/JCI92156

→ Emerging Targets in Photopharmacology; M. M. Lerch, M. J. Hansen, G. M. van Dam, W. Szymanski, B. L. Feringa; Angew. Chem. Int. Ed. 2016; 55: 10978-99. https://doi.org/10.1002/anie.201601931

→ Genetically encoding photoswitchable click amino acids in Escherichia coli and mammalian cells; C. Hoppmann, V. K. Lacey, G. V. Louie, J. Wei, J. P. Noel, L. Wang; Angew. Chem. Int. Ed. 2014; 53: 3932-6. https://doi.org/10.1002/anie.201400001

→ Reversible photocontrol of biological systems by the incorporation of molecular photoswitches; W. Szymanski, J. M. Beierle, H. A. Kistemaker, W. A. Velema, B. L. Feringa; Chem Rev 2013; 113: 6114-78. https://doi.org/10.1021/cr300179f

→ Photoisomerization in different classes of azobenzene; H. M. Bandara, S. C. Burdette; Chem Soc Rev 2012; 41: 1809-25. https://doi.org/10.1039/c1cs15179g

→ Azobenzene photoswitches for biomolecules; A. A. Beharry, G. A. Woolley; Chem Soc Rev 2011; 40: 4422-37. https://doi.org/10.1039/c1cs15023e

→ Properties of the pyramidal tract neuron system within the precentral wrist and hand area of primate motor cortex; D. R. Humphrey, W. S. Corrie, R. Rietz; J Physiol (Paris) 1978; 74: 215-26. https://doi.org/10.1016/j.isci.2021.102771

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8. Photocleavable Auxiliary Reagent for Native Chemical Ligation

Native Chemical Ligation is one of the most powerful tools for the preparation of complex peptides and small proteins. However, the classical variant of NCL requires an N-terminal cysteine at the ligation site. Iris Biotech presents an innovative auxiliary reagent for NCL that can be incorporated in place of a glycine residue.

Since glycine usually occurs several times in a peptide sequence, this approach significantly increases variability regarding the choice of possible ligation sites. In Native Chemical Ligation, the auxiliary’s SH-group mimics the action of an N-terminal cysteine’s sulfhydryl group.

Fig. 11: Native Chemical Ligation utilizing the photocleavable NCL-auxiliary glycine building block (PAA2000).

Following SPPS, the auxiliary is attached to the N-terminus of a peptide sequence in lieu of a glycine residue. The auxiliary’s Fmoc-protected amino functionality can subsequently be deprotected and functionalized, e.g. with a monodisperse PEG. PEGylation is useful for increasing the solubility of peptide fragments, and for facilitating their purification by precipitation with EtOH/Et 2 O. These properties are especially valuable if the peptide’s amino acid side chains are supposed to be further derivatized postSPPS, for example by enzymatic glycosylation. Following NCL, the auxiliary can be conveniently removed by irradiation with UV-light (10 min in water or water/acetonitrile). This method is particularly useful for the synthesis of sophisticated peptides such as glycopeptides, where cost- and labor-intensive short sequences can be prepared separately, and subsequently conjugated to long fragments synthesized in a standard manner.

PAA2000 t Bu-SS-Photo(Fmoc)-Gly-OH

Photocleavable-NCL-auxiliary-Gly-OH

CAS-No. 1994388-93-0

Formula C 36 H44N4O 9 S 2

Mol. weight 740,89 g/mol

Reference:

→ A PEGylated photocleavable auxiliary mediates the sequential enzymatic glycosylation and native chemical ligation of peptides; C. Bello, S. Wang, L. Meng, K. W. Moremen, C. F. Becker; Angew. Chem. Int. Ed. 2015; 54: 7711-5. https://doi.org/10.1002/anie.201501517

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Photochemistry

. 9. Photo-Linker for Solid Phase Synthesis of Peptide Amides and Acids

Peptide linkers are usually cleaved under acidic conditions or using two-step procedures. Photocleavage proceeds under neutral conditions using UV light and can either be performed in batch or using flow chemistry.

Furthermore, photolabile linkers are orthogonal to standard peptide chemistry reaction conditions, thus enabling the use of a wide variety of amino acid protecting groups.

Three different photolabile linkers are available for your convenience:

Photo-linker for the synthesis of C-terminal carboxylic acids

Fig. 12: Cleavage of photolabile linkers.

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Fmoc-amino photo-linker for the synthesis of peptide amides

RL-1026

Fmoc-Photo-Linker

4-{4-[1-(9-Fluorenylmethyloxycarbonylamino)ethyl]-2methoxy-5-nitrophenoxy}butanoic acid

CAS-No. 162827-98-7

Formula C 28 H28 N2O 8 Mol. weight 520,56 g/mol

BR-5205

Fmoc-Photolabile Resin 4-{4-[1-(Fmoc-amino)-ethyl]-2-methoxy-5-nitrophenoxy}butanamide polystyrene

References:

→ Continuous photochemical cleavage of linkers for solid-phase synthesis; M. Hurevich, J. Kandasamy, B. M. Ponnappa, M. Collot, D. Kopetzki, D. T. McQuade, P. H. Seeberger; Org Lett 2014; 16: 1794-7.

https://doi.org/10.1021/ol500530q

→ Photolytic Mass Laddering for Fast Characterization of Oligomers on Single Resin Beads; K. Burgess, C. I. Martinez, D. H. Russell, H. Shin, A. J. Zhang; J. Org. Chem. 1997; 62: 5662-5663.

https://doi.org/10.1021/jo970866w

→ Direct Monitoring of Organic Reactions on Polymeric Supports; M. R. Carrasco, M. C. Fitzgerald, Y. Oda, S. B. H. Kent; Tetrahedron Lett. 1997; 38: 6331-6334. https://doi.org/10.1016/s0040-4039(97)01456-1

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Photochemistry

10. Related Products

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10.1. Fmoc-Phe-Aca – a Cell Internalization Reporter, and further Fluorescent Amino Acid Derivatives

The Fmoc-L-Phe-Aca pseudo-dipeptide is a useful reporter group for the successful internalization of CPPs. Phe quenches the fluorescence of Aca. Internalization of the peptide containing Phe-Aca leads to proteolytic cleavage of the Phe-Aca bond and thus to fluorescence.

The unnatural amino acid Aca (7-amino-coumarin-4-acetic acid) is a coumarin derivative and thus exhibits fluorescence. When incorporated into a peptide C-terminally of phenylalanine, Aca is a useful reporter group for the successful internalization of CPPs. The phenyl moiety of Phe quenches the fluorescence of Aca. Internalization of the CPP containing Phe-Aca leads to proteolytic cleavage of the Phe-Aca bond and thus to fluorescence.

However, the peptide bond formation between Phe and Aca is considered to be a difficult coupling and often leads to low coupling yields. For your convenience, Iris Biotech offers Fmoc Phe-Aca as building block suitable for SPPS. This pseudodipeptide can be coupled to the resin of your choice and subsequently elongated to prepare your target cell penetrating peptides.

Fig. 13: Loading of a solid support, subsequent elongation to CPPs and function as a reporter group for successful CPP internalization.

In addition to Fmoc-L-Phe-Aca-OH, we offer various 7-amino-4-methylcoumarin (AMC)-functionalized amino acid derivatives. Like Aca, when bound to a peptide, the AMC-amide fluoresces very weakly and excitation/emission wavelengths are shorter (ca. 330/390 nm). When the free AMC-amine is released by proteolytic cleavage, the fluorescence increases by a factor of approx. 700 and excitation and emission wavelengths are red-shifted. Thus, the low fluorescence of the AMC-amide substrate does not interfere with the fluorometric assay. Besides, Iris Biotech offers dipeptide recognition motifs such as Phe-Arg (ZAA1480) or Arg-Arg (ZAA1470) as fluorogenic 7-amido-4-methylcoumarin based protease substrates. The acetic acid derivative of AMC, 7-amino-4-methyl coumarin-3-acetic acid (AMCA) emits in the blue region (440-460 nm) upon activation with UV light of 350 nm.

Further fluorescent amino acids include those functionalized with acridine derivatives (2-Acd and 2-Bacd).

Coumarin Derivatives

BAA3650

Boc-L-Lys-AMC*AcOH

N-alpha-t-Butyloxycarbonyl-L-lysine 7-amido-4-methylcoumarin acetate

CAS-No. 116883-12-6 net

Formula C 21H29 N3 O 5*CH3 CO 2H

Mol. weight 403,47*60,05

FDP1240 Fmoc-L-Phe-Aca-OH

7-[N-alpha-(9-Fluorenylmethyloxycarbonyl)-L-phenylalaninyl-amido]-coumarin-4-acetic acid

CAS-No. 2250437-40-0

Formula C 35H28 N2O 7

Mol. weight 588,61 g/mol

HAA1200 H-L-Ala-AMC

L-Alanine-7-amido-4-methylcoumarin

CAS-No. 77471-41-1

Formula C13 H14N2O 3

Mol. weight 246,27 g/mol

HAA7760 H-L-Thr-AMC

L-Threonine 7-amido-4-methylcoumarin, 98%

CAS-No. 191723-66-7

Formula C14H16 N2O4

Mol. weight 276,29 g/mol back to content

Photochemistry

HAA1174 H-L-Ala-AMC*TFA

L-Alanine 7-amido-4-methylcoumarin trifluoroacetate

CAS-No. 96594-10-4

Formula C13 H14N2O 3*CF 3 CO 2H

Mol. weight 360,29 g/mol

HAA7630 H-L-Arg-AMC*2HCl

L-Arginine 7-amido-4-methylcoumarin dihydrochloride

CAS-No. 113712-08-6

Formula C16 H21N 5 O 3*2HCl

Mol. weight 331,37*72,90 g/mol

HAA7645 Glt-Gly-Arg-AMC.HCl

Glutaryl-glycyl-L-arginine 7-amido-4-methylcoumarin hydrochloride

CAS-No. 103213-40-7

Formula C 23 H30 ClN 6 O 7 *HCl

Mol. weight 502,53*36,46 g/mol

HAA3200 H-L-Leu-AMC

L-Leucine-7-amido-4-methylcoumarine

CAS-No. 66447-31-2

Formula C16 H20 N2O 3

Mol. weight 288,34 g/mol

HAA7740 H-L-Pyr-AMC

L-Pyroglutamic acid 7-amido-4-methylcoumarin

CAS-No. 66642-36-2

Formula C15H14N2O4

Mol. weight 286,29 g/mol

HAA2890 H-L-Asp(AMC)-OH

L-Aspartic acid beta-(7-amido-4-methylcoumarin)

CAS-No. 133628-73-6

Formula C14H14N2O 5

Mol. weight 290,27 g/mol

HAA7971 H-Gly-AMC

Glycine 7-amido-4-methylcoumarin

CAS-No. 77471-42-2

Formula C12H12N2O 3

Mol. weight 232,24 g/mol

RL-1005 AMCA-OSu

7-Amino-4-methyl-3-coumarinylacetyl succinimidyl ester AMCO-NHS

CAS-No. 113721-87-2

Formula C16 H14N2O 6

Mol. weight 330,29 g/mol

RL-1170

Fmoc-ACA-OH

7-(9-Fluorenylmethyloxycarbonylamino)-coumarin-4-acetic acid

CAS-No. 378247-75-7

Formula C 26 H19 NO 6

Mol. weight 441,43 g/mol

ZAA1470

Z-L-Arg-L-Arg-AMC*2HCl

N-alpha-benzyloxycarbonyl-L-arginyl-L-arginine-7-amido-4-methylcoumarin-dihydrochloride

CAS-No. 140686-23-3

Formula C 30 H39 N9 O 6*2HCl

Mol. weight 621,69*72,91 g/mol

ZAA1480

Z-L-Phe-L-Arg-AMC*HCl

N-alpha-benzyloxycarbonyl-L-phenylalanyl-L-arginine-7-amido-4-methylcoumarin hydrochlorid

CAS-No. 65147-22-0

Formula C 33 H36 N 6 O 6*HCl

Mol. weight 612,69*36,46 g/mol

Photochemistry

Acridine Derivatives

BAA1580 Boc-L-Ala(2-Bacd)-OH

N-alpha-t-Butyloxycarbonyl-3-[benzo[b]acridin-12(5H)on-2-yl]-L-alanine

CAS-No. 916834-72-5

Formula C 25H24N2O 5 Mol. weight 432,47 g/mol

BAA1590 Boc-L-Ala(2-Acd)-OH

N-alpha-t-Butyloxycarbonyl-3-{2-[9(10H)-acridonyl]}-L-alanine

CAS-No. 643018-86-4

Formula C 21H22N

FAA5830 Fmoc-L-Ala(2-Acd)-OH

N-alpha-(9-Fluorenylmethoxycarbonyl)-3-{2-[9(10H)-acridonyl]}-L-alanine

CAS-No. 1013328-63-6

Formula C 31H24N2O 5 Mol. weight 504,53 g/mol

FAA5880 Fmoc-L-Ala(2-Bacd)-OH

N-alpha-(9-Fluorenylmethoxycarbonyl)-3-[benzo[b] acridin-12(5H)-on-2-yl]-L-alanine

CAS-No. 1157859-85-2

Formula C 35H26 N2O 5 Mol. weight 554,59 g/mol

HAA3570 H-L-Ala(2-Bacd)-OH*HCl

3-[Benzo[b]acridin-12(5H)-on-2-yl]-L-alanine hydrochloride

Formula C 20 H16 N2O 3*HCl Mol.

HAA3580 H-L-Ala(2-Acd)-OH*HCl

3-{2-[9(10H)-acridonyl]}-L-alanine hydrochloride

CAS-No. 854503-32-5 net

Formula C16 H14N2O 3*HCl

Mol. weight 282,29*36,45 g/mol

References:

→ Rapid and general profiling of protease specificity by using combinatorial fluorogenic substrate libraries; J. L. Harris, B. J. Backes, F. Leonetti, S. Mahrus, J. A. Ellman, C. S. Craik; PNAS 2000; 97: 7754-7759. https://doi.org/10.1073/pnas.140132697

→ Expedient Solid-Phase Synthesis of Fluorogenic Protease Substrates Using the 7-Amino-4-carbamoylmethylcoumarin (ACC) Fluorophore; D. J. Maly, F. Leonetti, B. J. Backes, D. S. Dauber, J. L. Harris, C. S. Craik, J. A. Ellman; J. Org. Chem. 2002; 67: 910-915. https://doi.org/10.1021/jo016140o

→ Synthesis of a New Fluorogenic Substrate for Cystine Aminopeptidase; Y. Kanaoka, T. Takahashi, H. Nakayama, T. Ueno, T. Sekine; Chem. Pharm. Bull. 1982; 30(40) : 1485-1487. https://doi.org/10.1248/cpb.30.1485

→ Sensitive assays for trypsin, elastase, and chymotrypsin using new fluorogenic substrates; M. Zimmerman, B. Ashe, E. C. Yurewicz, G. Patel; Anal. Biochem. 1977; 78(1): 47-51. https://doi.org/10.1016/0003-2697(77)90006-9.

→ Aminomethyl coumarin acetic acid: a new fluorescent labelling agent for proteins; H. Khalfan, R. Abuknesha, M. Rand-Weaver, R. G. Price, D. Robinson; Histochem J. 1986; 18(9) : 497-499.

https://doi.org/10.1007/BF01675617

→ A Simple Fluorescent Labeling Method for Studies of Protein Oxidation, Protein Modification, and Proteolysis; A. M. Pickering, K. J. A. Davies; Free Radic Biol Med. 2012; 52(2) : 239-246.

https://doi.org/10.1016/j.freeradbiomed.2011.08.018

→ Fluorescence imaging of drug target proteins using chemical probes; H. Zhu, I. Hamachi; J. Pharm. Anal. 2020; 10: 426-433. https://doi.org/10.1016/j.jpha.2020.05.013

→ A New Fluorogenic Substrate for Chymotrypsin; M. Zimmerman, E. Yurewicz, G. Patel; Anal. Biochem. 1976; 70: 258-262. https://doi.org/10.1016/S0003-2697(76)80066-8

→ Determination of Caspase Specificities Using a Peptide Combinatorial Library; N. A. Thornberry, K. T. Chapman, D. W. Nicholson; Method Enzymol 2000; 322: 100-110.

https://doi.org/10.1016/S0076-6879(00)22011-9

Photochemistry

10.2. Building Blocks for the SPPS of FRET-based fluorogenic protease substrates

The classic FRET pair EDANS and DABCYL is now available linked to Fmoc amino acid building blocks that can be readily used in SPPS. Conveniently synthesize your own custom protease substrates!

Peptide

Fragment 1

Traditionally, fluorogenic protease substrates for the screening of protease activity are prepared by peptide synthesis and subsequent regioselective deprotection and functionalization with a fluorophore/ quencher pair such as EDANS and DABSYL. For your convenience, we now offer a glutamate and a lysine building block functionalized with EDANS and DABSYL, respectively. Those building blocks are suitable for peptide synthesis using the Fmoc strategy and will allow you to prepare custom protease substrates without additional tedious and yield-reducing functionalization steps post-SPPS.

Product details

FAA1498 Fmoc-L-Lys(Dabcyl)-OH

N-alpha-(9-Fluorenylmethyloxycarbonyl)-N-epsilon-4-[4‘-(dimethylamino)phenylazo]benzoyl-L-lysine

CAS-No. 146998-27-8

Formula C 36 H37N 5 O 5

Mol. weight 619,73 g/mol

LS-4180 Dabcyl-KTSAVLQSGFRKM-Glu(Edans)-amide

Dabcyl-lysyl-threonyl-seryl-alanyl-valyl-leucyl-glutaminyl-seryl-glycyl-phenylalanyl-arginyl-lysyl-methionyl-glutamyl(Edans)-amide

CAS-No. 2642630-08-6

Formula C95H142N26 O 23 S 2

Mol. weight 2080,46 g/mol

Fig. 14: Screening of protease activity using FRET-based fluorogenic protease substrates

FAA4410 Fmoc-L-Glu(EDANS)-OH N-alpha-(9-Fluorenylmethyloxycarbonyl)-5-(2-(5-sulfonaphthalen-1-ylamino)ethylamido)-L-glutamic acid

CAS-No. 193475-66-0

Formula C 32H31N3 O 8 S

Mol. weight 617,67 g/mol

References:

→ Enzymatic activity characterization of SARS coronavirus 3C-like protease by fluorescence resonance energy transfer technique; S. Chen, L. L. Chen, H. B. Luo, T. Sun, J. Chen, F. Ye, J. H. Cai, J. K. Shen, X. Shen, H. L. Jiang; Acta Pharmacol Sin 2005; 26: 99-106. https://doi.org/10.1111/j.1745-7254.2005.00010.x

→ Synthesis and evaluation of fluorescent probes for the detection of calpain activity; S. Mittoo, L. E. Sundstrom, M. Bradley; Anal Biochem 2003; 319: 234-8. https://doi.org/10.1016/s0003-2697(03)00324-5

→ A general method for the preparation of internally quenched fluorogenic protease substrates using solid-phase peptide synthesis; L. L. Maggiora, C. W. Smith, Z. Y. Zhang; J Med Chem 1992; 35: 3727-30. https://doi.org/10.1021/jm00099a001

→ Novel fluorogenic substrates for assaying retroviral proteases by resonance energy transfer; E. D. Matayoshi, G. T. Wang, G. A. Krafft, J. Erickson; Science 1990; 247: 954-8. https://doi.org/10.1126/science.2106161

→ Design and synthesis of new fluorogenic HIV protease substrates based on resonance energy transfer; G. T. Wang, E. Matayoshi, H. Jan Huffaker, G. A. Krafft; Tetrahedron Lett. 1990; 31: 6493-6496. https://doi.org/10.1016/s0040-4039(00)97099-0

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Photochemistry Code of Conduct

As business activity of Iris Biotech GmbH impacts people’s lives and health, it must be operated in ethical and correct manner and act with integrity and responsibility. To ensure high ethical standards and fair business practices, Iris Biotech GmbH applies an integrated policy known as its Code of Conduct.

In 2001 Iris Biotech GmbH was founded just at the beginning of the Biotech movement and the first remarkable breakthrough of biotech pharma products. Although the biotech field is rather young compared to other industries we believe on long-term business, a good partnership between our business partners and Iris Biotech GmbH and a good reputation. It is our duty as well as our responsibility to maintain and to extend this over the next generations – based on the principles of an honourable and prudent tradesman which based upon the concept of honourable entrepreneurship.

This Code of Conduct has been developed following the “Voluntary Guidelines for Manufacturers of Fine Chemical Intermediates and Active Ingredients” issued by AIME (Agrochemical & Intermediates Manufacturers in Europe) and the requirements of some of our business associates.

Iris Biotech GmbH commits to hold this Code of Conduct and to include and apply its principles in the management system and the company policies.

Ethics

Iris Biotech GmbH undertakes business in an ethical manner and acts with integrity. All corruption, extortion and embezzlement are prohibited. We do not pay or accept bribes or participate in other illegal inducements in business or government relationships. We conduct our business in compliance with all applicable anti-trust laws. Employees are encouraged to report concerns or illegal activities in the workplace, without threat of reprisal, intimidation or harassment.

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General Policies

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Iris Biotech GmbH provides a safe and healthy working environment to the employees and protects them from overexposure to chemical and physical hazards. Products are produced, stored and shipped under the guidelines of the relevant chemical and safety legislation. Risks and emergency scenarios are identified and evaluated, and their possible impact is minimized by implementing emergency plans and written procedures. Safety information regarding hazardous materials is available to educate, train and protect workers from hazards. Preventive equipment and facilities maintenance is performed at suitable periods to reduce potential hazards. Employees are regularly trained in health and safety matters and are informed about product properties and risk classification when it is required.

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A quality management system following the Good Distribution Practices (GDP rules) of Active Pharmaceutical Ingredients is established covering all the aspects of the worldwide distribution of products. Regular audits are performed to evaluate the efficiency and fulfilling of the quality system. Process controls to provide reproducible product quality are established. There are preventive maintenance procedures to ensure plant reliability and the lowest risk of failure. Staff is trained periodically about GMP and GDP rules. Procedures are established and installations are designed to avoid cross contamination. Batch and analytical records are kept for inspection and audit purposes for suitable periods according guidelines.

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Photochemistry Terms and Conditions of Sales

All orders placed by a buyer are accepted and all contracts are made subject to the terms which shall prevail and be effective notwithstanding any variations or additions contained in any order or other document submitted by the buyer. No modification of these terms shall be binding upon Iris Biotech GmbH unless made in writing by an authorised representative of Iris Biotech GmbH.

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Every order made by the buyer shall be deemed an offer by the buyer to purchase products from Iris Biotech GmbH and will not be binding on Iris Biotech GmbH until a duly authorised representative of Iris Biotech GmbH has accepted the offer made by the buyer. Iris Biotech GmbH may accept orders from commercial, educational or government organisations, but not from private individuals and Iris Biotech GmbH reserves the right to insist on a written order and/or references from the buyer before proceeding.

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Prices are subject to change. For the avoidance of doubt, the price advised by Iris Biotech GmbH at the time of the buyer placing the order shall supersede any previous price indications. The buyer must contact the local office of Iris Biotech GmbH before ordering if further information is required. Unless otherwise agreed by the buyer and Iris Biotech GmbH, the price shall be for delivery ex-works. In the event that the buyer requires delivery of the products otherwise than ex-works the buyer should contact the local office of Iris Biotech GmbH in order to detail its requirements. Iris Biotech GmbH shall, at its discretion, arrange the buyer‘s delivery requirements including, without limitation, transit insurance, the mode of transit (Iris Biotech GmbH reserves the right to vary the mode of transit if any regulations or other relevant considerations so require) and any special packaging requirements (including cylinders). For the avoidance of doubt all costs of delivery and packaging in accordance with the buyer‘s requests over and above that of delivery in standard packaging ex-works shall be for the buyer‘s account unless otherwise agreed by both parties. Incoterms 2020 shall apply. Any tax, duty or charge imposed by governmental authority or otherwise and any other applicable taxes, duties or charges shall be for the buyer‘s account. Iris Biotech GmbH may, on request and where possible, provide quotations for multiple packs or bulk quantities, and non-listed items. Irrespective of the type of request or means of response all quotations must be accepted by the buyer without condition and in writing before an order will be accepted by Iris Biotech GmbH. Unless agreed in writing on different terms, quotations are valid for 30 days from the date thereof. Payment terms are net 30 days from invoice date unless otherwise agreed in writing. Iris Biotech GmbH reserves the right to request advance payment at its discretion. For overseas transactions the buyer shall pay all the banking charges of Iris Biotech GmbH. The buyer shall not be entitled to withhold or set-off payment for the products for any reason whatsoever. Government/

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The buyer shall inspect goods immediately on receipt and inform Iris Biotech GmbH of any shortage or damage within five days. Quality problems must be notified within ten days of receipt. Goods must not be returned without prior written authorisation of Iris Biotech GmbH. Iris Biotech GmbH shall at its sole discretion replace the defective products (or parts thereof) free of charge or refund the price (or proportionate price) to buyer. Opened or damaged containers cannot be returned by the buyer without the written prior agreement of Iris Biotech GmbH. In the case of agreed damaged containers which cannot be so returned, the buyer assumes responsibility for the safe disposal of such containers in accordance with all applicable laws.

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Products are analysed in the Quality Control laboratories of Iris Biotech GmbH’s production partners by methods and procedures which Iris Biotech GmbH considers appropriate. In the event of any dispute concerning reported discrepancies arising from the buyer’s analytical results, determined by the buyer’s own analytical procedures, Iris Biotech GmbH reserves the right to rely on the results of own analytical methods of Iris Biotech GmbH. Certificates of Analysis or Certificates of Conformity are available at the discretion of Iris Biotech GmbH for bulk orders but not normally for prepack orders. Iris Biotech GmbH reserves the right to make a charge for such certification. Specifications may change and reasonable variation from any value listed should not form the basis of a dispute. Any supply by Iris Biotech GmbH of bespoke or custom product for a buyer shall be to a specification agreed by both parties in writing. Technical information, provided orally, in writing, or by electronic means by or on behalf of Iris Biotech GmbH, including any descriptions, references, illustrations or diagrams in any catalogue or brochure, is provided for guidance purposes only and is subject to change.

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Photochemistry

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All products of Iris Biotech GmbH are intended for laboratory research purposes and unless otherwise stated on product labels, in the catalogue and product information sheet of Iris Biotech GmbH or in other literature furnished to the buyer, are not to be used for any other purposes, including but not limited to use as or as components in drugs for human or animal use, medical devices, cosmetics, food additives, household chemicals, agricultural or horticultural products or pesticides. Iris Biotech GmbH offers no warranty regarding the fitness of any product for a particular purpose and shall not be responsible for any loss or damage whatsoever arising there from. No warranty or representation is given by Iris Biotech GmbH that the products do not infringe any letters patent, trademarks, registered designs or other industrial rights. The buyer further warrants to Iris Biotech GmbH that any use of the products in the United States of America shall not result in the products becoming adulterated or misbranded within the meaning of the Federal Food, Drug and Cosmetic Act (or such equivalent legislation in force in the buyer‘s jurisdiction) and shall not be materials which may not, under sections 404, 505 or 512 of the Act, be introduced into interstate commerce. The buyer acknowledges that, since the products of Iris Biotech GmbH are intended for research purposes, they may not be on the Toxic Substances Control Act 1976 („TSCA“) inventory. The buyer warrants that it shall ensure that the products are approved for use under the TSCA (or such other equivalent legislation in force in the buyer‘s jurisdiction), if applicable. The buyer shall be responsible for complying with any legislation or regulations governing the use of the products and their importation into the country of destination (for the avoidance of doubt to include, without limitation, the TSCA and all its amendments, all EINECS, ELINCS and NONS regulations). If any licence or consent of any government or other authority shall be required for the acquisition, carriage or use of the products by the buyer the buyer shall obtain the same at its own expense and if necessary produce evidence of the same to Iris Biotech GmbH on demand. Failure to do so shall not entitle the buyer to withhold or delay payment. Any additional expenses or charges incurred by Iris Biotech GmbH resulting from such failure shall be for the buyer‘s account. Save for death or personal injury caused by negligence of Iris Biotech GmbH, sole obligation of Iris Biotech GmbH and buyer‘s exclusive remedy with respect to the products proved to the satisfaction of Iris Biotech GmbH to be defective or products incorrectly supplied shall be to accept the return of said products to Iris Biotech GmbH for refund of the actual purchase price paid by the buyer (or proportionate part thereof), or replacement of the defective product (or part thereof) with alternative product. Iris Biotech GmbH shall have no liability to the buyer under or arising directly or indirectly out of or otherwise in connection with the supply of products by Iris Biotech GmbH to the buyer and/or their re-sale or use by the buyer or for any product, process or services of the buyer which in any way comprises the product in contract tort (including negligence or breach of statutory duty) or otherwise for pure economic loss, loss of profit, business, reputation, depletion of brand, contracts, revenues or anticipated savings or for any special indirect or consequential damage or loss of any nature except as may otherwise be expressly provided for in these terms. All implied warranties, terms and representations in respect of the products (whether implied by statute or otherwise) are excluded to the fullest extent permitted by law. The buyer shall indemnify Iris Biotech GmbH for and against any and all losses, damages and expenses, including legal fees and other costs of defending any action, that Iris Biotech GmbH may sustain or incur as a result of any act or omission by the buyer, its officers, agents or employees, its successors or assignees, its customers or all other third parties, whether direct or indirect, in connection with the use of any product. For the avoidance of doubt and in the event that Iris Biotech GmbH supplies bespoke or custom product to the buyer‘s design or specification, this indemnity shall extend to include any claim by a third party that the manufacture of the product for the buyer or the use of the product by the buyer infringes the intellectual property rights of any third party.

General

Iris Biotech GmbH shall be entitled to assign or sub-contract all or any of its rights and obligations hereunder. The buyer shall not be entitled to assign, transfer, sub-contract or otherwise delegate any of its rights or obligations hereunder. Any delay or forbearance by Iris Biotech GmbH in exercising any right or remedy under these terms shall not constitute a waiver of such right or remedy. If any provision of these terms is held by any competent authority to be invalid or unenforceable in whole or in part the validity of the other provisions of these terms and the remainder of the provision in question shall not be affected. These terms shall be governed by German Law and the German Courts shall have exclusive jurisdiction for the hearing of any dispute between the parties save in relation to enforcement where the jurisdiction of the German Courts shall be non-exclusive.

Notes

Photochemistry Index

Get in Contact

Iris Biotech GmbH

Adalbert-Zoellner-Str. 1 95615 Marktredwitz Germany

+49 (0) 9231 97121-0 +49 (0) 9231 97121-99 info@iris-biotech.de www.iris-biotech.de

Distribution Partners

The list contains the current distributors of Iris Biotech in different regions of the world. The latest list of distribution partners and contact details is available at: www.iris-biotech.de/distribution-partner

China:

Chengdu Yoo Technology Co., Ltd.

Japan:

BizCom Japan, Inc.

Shigematsu & Co., Ltd

Cosmo Bio Co., Ltd.

USA & Canada: Peptide Solutions LLC

India, Bangladesh, Oman, Sri Lanka, United Arab Emirates: Sumit Biosciences Pvt Ltd.

Singapore, Thailand, Malaysia, Indonesia, Vietnam, Philippines, Brunei, Burma (Myanmar), Laos, Cambodia, Timor-Leste:

SciClix Pte. Ltd.

Empowering Peptide Innovation