813 results on '"Duocarmycins"'
Search Results
2. Recent Progress on the Total Synthesis of Duocarmycins A and SA, Yatakemycin, and PDE-I and PDE-II
- Author
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Sakata, Juri, Tokuyama, Hidetoshi, and Kobayashi, Yuichi, editor
- Published
- 2019
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3. The DNA glycosylase AlkD uses a non-base-flipping mechanism to excise bulky lesions
- Author
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Mullins, Elwood A, Shi, Rongxin, Parsons, Zachary D, Yuen, Philip K, David, Sheila S, Igarashi, Yasuhiro, and Eichman, Brandt F
- Subjects
Inorganic Chemistry ,Chemical Sciences ,Genetics ,Bacillus cereus ,Base Pairing ,Biocatalysis ,Catalytic Domain ,Crystallography ,X-Ray ,DNA Adducts ,DNA Damage ,DNA Glycosylases ,DNA Repair ,Duocarmycins ,Indoles ,Models ,Molecular ,Pyrroles ,General Science & Technology - Abstract
Threats to genomic integrity arising from DNA damage are mitigated by DNA glycosylases, which initiate the base excision repair pathway by locating and excising aberrant nucleobases. How these enzymes find small modifications within the genome is a current area of intensive research. A hallmark of these and other DNA repair enzymes is their use of base flipping to sequester modified nucleotides from the DNA helix and into an active site pocket. Consequently, base flipping is generally regarded as an essential aspect of lesion recognition and a necessary precursor to base excision. Here we present the first, to our knowledge, DNA glycosylase mechanism that does not require base flipping for either binding or catalysis. Using the DNA glycosylase AlkD from Bacillus cereus, we crystallographically monitored excision of an alkylpurine substrate as a function of time, and reconstructed the steps along the reaction coordinate through structures representing substrate, intermediate and product complexes. Instead of directly interacting with the damaged nucleobase, AlkD recognizes aberrant base pairs through interactions with the phosphoribose backbone, while the lesion remains stacked in the DNA duplex. Quantum mechanical calculations revealed that these contacts include catalytic charge-dipole and CH-π interactions that preferentially stabilize the transition state. We show in vitro and in vivo how this unique means of recognition and catalysis enables AlkD to repair large adducts formed by yatakemycin, a member of the duocarmycin family of antimicrobial natural products exploited in bacterial warfare and chemotherapeutic trials. Bulky adducts of this or any type are not excised by DNA glycosylases that use a traditional base-flipping mechanism. Hence, these findings represent a new model for DNA repair and provide insights into catalysis of base excision.
- Published
- 2015
4. Duocarmycin-based antibody–drug conjugates as an emerging biotherapeutic entity for targeted cancer therapy: Pharmaceutical strategy and clinical progress.
- Author
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Yao, Hang-Ping, Zhao, Hui, Hudson, Rachel, Tong, Xiang-Min, and Wang, Ming-Hai
- Subjects
- *
ANTIBODY-drug conjugates , *TARGETED drug delivery , *CANCER treatment - Abstract
• Duocarmycins and analogues are DNA-alkylators highly cytotoxic to cancer cells. • Up to now, none of duocarmycins have not been approved for oncological practice. • Novel features make duocarmycins an ideal payload for targeted drug delivery. • Various duocarmycin-based antibody-drug conjugates are in clinical development. • An emerging era of duocarmycin-based anticancer biotherapeutics is in the horizon. Duocarmycins are a class of DNA minor-groove-binding alkylating molecules. For the past decade, various duocarmycin analogues have been used as payloads in the development of antibody–drug conjugates (ADCs). Currently, more than 15 duocarmycin-based ADCs have been studied preclinically, and some of them such as SYD985 have been granted Fast-Track Designation status. Nevertheless, progress in duocarmycin-based ADCs also faces challenges, with setbacks including the termination of BMS-936561/MDX-1203. In this review, we discuss issues associated with the efficacy, pharmacokinetic profile, and toxicological activity of these biotherapeutics. Furthermore, we summarize the latest advances in duocarmycin-based ADCs that have different target specificities and linker chemistries. Evidence from preclinical and clinical studies has indicated that duocarmycin-based ADCs are promising biotherapeutics for oncological application in the future. [ABSTRACT FROM AUTHOR]
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- 2021
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5. Antibody-mimetic drug conjugate with efficient internalization activity using anti-HER2 VHH and duocarmycin.
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Sakata J, Tatsumi T, Sugiyama A, Shimizu A, Inagaki Y, Katoh H, Yamashita T, Takahashi K, Aki S, Kaneko Y, Kawamura T, Miura M, Ishii M, Osawa T, Tanaka T, Ishikawa S, Tsukagoshi M, Chansler M, Kodama T, Kanai M, Tokuyama H, and Yamatsugu K
- Subjects
- Humans, Duocarmycins, Receptor, ErbB-2 genetics, Cell Line, Tumor, Drug Delivery Systems, Immunoconjugates pharmacology
- Abstract
Antibody-mimetic drug conjugate (AMDC) is a cancer cell-targeted drug delivery system based on the non-covalent binding of mutated streptavidin and modified biotin, namely Cupid and Psyche. However, the development of AMDCs is hampered by difficulties in post-translational modification or poor internalization activity. Here, we report an expression, refolding, and purification method for AMDC using a variable heavy chain of heavy chain-only antibodies (VHHs). Monomeric anti-HER2 VHH fused to Cupid was expressed in Escherichia coli inclusion bodies. Solubilization and refolding at optimized reducing conditions and pH levels were selected to form a functional, tetrameric protein (anti-HER2 VHH-Cupid) that can be easily purified based on molecular weight. Anti-HER2 VHH-Cupid non-covalently creates a tight complex with Psyche linked to a potent DNA-alkylating agent, duocarmycin. This complex can be absorbed by the HER2-expressing human breast cancer cell line, KPL-4, and kills KPL-4 cells in vitro and in vivo. The production of a targeting protein with internalizing activity, combined with the non-covalent conjugation of a highly potent payload, renders AMDC a promising platform for developing cancer-targeted therapy., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
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- 2024
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6. Generation and Characterization of Iduronidase-Cleavable ADCs.
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Jäger S, Könning D, Rasche N, Hart F, Sensbach J, Krug C, Raab-Westphal S, Richter K, Unverzagt C, Hecht S, Anderl J, and Schröter C
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- Iduronidase, Duocarmycins, Antibodies, Monoclonal, Cell Line, Tumor, Immunoconjugates metabolism, Antineoplastic Agents pharmacology, Antineoplastic Agents therapeutic use, Antineoplastic Agents metabolism
- Abstract
A crucial design feature for the therapeutic success of antibody-drug conjugates (ADCs) is the linker that connects the antibody with the drug. Linkers must be stable in circulation and efficiently release the drug inside the target cell, thereby having a fundamental impact on ADC pharmacokinetics and efficacy. The variety of enzymatically cleavable linkers applied in ADCs is limited, and some are believed to be associated with unwanted side effects due to the expression of cleavage-mediating enzymes in nonmalignant cells. Based on a bioinformatic screen of lysosomal enzymes, we identified α-l-iduronidase (IduA) as an interesting candidate for ADC linker cleavage because of its low expression in normal tissues and its overexpression in several tumor types. In the present study, we report a novel IduA-cleavable ADC linker using exatecan and duocarmycin as payloads. We showed the functionality of our linker system in cleavage assays using recombinant IduA or cell lysates and compared it to established ADC linkers. Subsequently, we coupled iduronide-exatecan via interchain cysteines or iduronide-duocarmycin via microbial transglutaminase (mTG) to an anti-CEACAM5 (aCEA5) antibody. The generated iduronide-exatecan ADC showed high serum stability and similar target-dependent tumor cell killing in the subnanomolar range but reduced toxicity on nonmalignant cells compared to an analogous cathepsin B-activatable valine-citrulline-exatecan ADC. Finally, in vivo antitumor activity could be demonstrated for an IduA-cleavable duocarmycin ADC. The presented results emphasize the potential of iduronide linkers for ADC development and represent a tool for further balancing out tumor selectivity and safety.
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- 2023
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7. Novel Thienoduocarmycin-Trastuzumab ADC Demonstrates Strong Antitumor Efficacy with Favorable Safety Profile in Preclinical Studies.
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Valsasina B, Orsini P, Caruso M, Albanese C, Ciavolella A, Cucchi U, Fraietta I, Melillo N, Fiorentini F, Rizzi S, Salsa M, Isacchi A, and Gasparri F
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- Humans, Mice, Animals, Duocarmycins, Macaca fascicularis metabolism, Receptor, ErbB-2 metabolism, Cell Line, Tumor, Trastuzumab pharmacology, Trastuzumab therapeutic use, Xenograft Model Antitumor Assays, Antineoplastic Agents pharmacology, Antineoplastic Agents therapeutic use, Antineoplastic Agents chemistry, Immunoconjugates pharmacology, Immunoconjugates therapeutic use, Immunoconjugates chemistry
- Abstract
New antibodies-drug conjugate (ADC) payloads overcoming chemoresistance and killing also poorly proliferating tumors at well-tolerated doses are much desired. Duocarmycins are a well-known class of highly potent cytotoxic agents, with DNA minor groove-binding and alkylation properties, active also in chemoresistant tumors. Although different duocarmycin derivatives have been used during the years as payloads for ADC production, unfavorable physicochemical properties impaired the production of ADCs with optimal features. Optimization of the toxin to balance reactivity and stability features and best linker selection allowed us to develop the novel duocarmycin-like payload-linker NMS-P945 suitable for conjugation to mAbs with reproducible drug-antibody ratio (DAR) >3.5. When conjugated to trastuzumab, it generated an ADC with good internalization properties, ability to induce bystander effect and immunogenic cell death. Moreover, it showed strong target-driven activity in cells and cytotoxic activity superior to trastuzumab deruxtecan tested, in parallel, in cell lines with HER2 expression. High in vivo efficacy with cured mice at well-tolerated doses in HER2-driven models was also observed. A developed pharmacokinetic/pharmacodynamic (PK/PD) model based on efficacy in mice and cynomolgus monkey PK data, predicted tumor regression in patients upon administration of 2 doses of trastuzumab-NMS-P945-ADC at 0.5 mg/kg. Thus, considering the superior physicochemical features for ADC production and preclinical results obtained with the model trastuzumab ADC, including bystander effect, immunogenic cell death and activity in chemoresistant tumors, NMS-P945 represents a highly effective, innovative payload for the creation of novel, next-generation ADCs., (©2023 American Association for Cancer Research.)
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- 2023
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8. Antibody-drug Conjugate PCMC1D3-Duocarmycin SA as a Novel Therapeutic Entity for Targeted Treatment of Cancers Aberrantly Expressing MET Receptor Tyrosine Kinase
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Rachel Hudson, Hang-Ping Yao, Sreedhar Reddy Suthe, Dhavalkumar Patel, and Ming-Hai Wang
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Pharmacology ,Cancer Research ,Immunoconjugates ,Body Weight ,Proto-Oncogene Proteins c-met ,Xenograft Model Antitumor Assays ,Duocarmycins ,Mice ,Oncology ,Cell Line, Tumor ,Neoplasms ,Drug Discovery ,Animals ,Humans - Abstract
Background: Aberrant expression of the MET receptor tyrosine kinase is an oncogenic determinant and a drug target for cancer therapy. Currently, antibody-based biotherapeutics targeting MET are under clinical trials. Objective: Here, we report the preclinical and therapeutic evaluation of a novel anti-MET antibody- drug conjugate PCMC1D3-duocarmycin SA (PCMC1D3-DCM) for targeted cancer therapy. Methods: The monoclonal antibody PCMC1D3 (IgG1a/κ), generated by a hybridoma technique and specific to one of the MET extracellular domains, was selected based on its high specificity to human MET with a binding affinity of 1.60 nM. PCMC1D3 was conjugated to DCM via a cleavable valine-citrulline dipeptide linker to form an antibody-drug conjugate with a drug-to-antibody ratio of 3.6:1. PCMC1D3-DCM in vitro rapidly induced MET internalization with an internalization efficacy ranging from 6.5 to 17.2h dependent on individual cell lines. Results: Studies using different types of cancer cell lines showed that PCMC1D3-DCM disrupted the cell cycle, reduced cell viability, and caused massive cell death within 96h after treatment initiation. The calculated IC50 values for cell viability reduction were 1.5 to 15.3 nM. Results from mouse xenograft tumor models demonstrated that PCMC1D3-DCM in a single dose injection at 10 mg/kg body weight effectively delayed xenograft tumor growth up to two weeks without signs of tumor regrowth. The calculated tumoristatic concentration, a minimal dose required to balance tumor growth and inhibition, was around 2 mg/kg body weight. Taken together, PCMC1D3-DCM was effective in targeting the inhibition of tumor growth in xenograft models. Conclusion: This work provides the basis for the development of humanized PCMC1D3-DCM for MET-targeted cancer therapy in the future.
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- 2022
9. Excision repair of adozelesin-N3 adenine adduct by 3-methyladenine-DNA glycosylases and UvrABC nuclease.
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Jin, SG, Choi, JH, Ahn, B, O'Connor, TR, Mar, W, and Lee, CS
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Biochemistry and Cell Biology ,Biological Sciences ,Genetics ,Cancer ,Underpinning research ,1.1 Normal biological development and functioning ,Alkylation ,Animals ,Antineoplastic Agents ,Alkylating ,Bacterial Proteins ,Benzofurans ,Cyclohexanecarboxylic Acids ,Cyclohexenes ,DNA Adducts ,DNA Glycosylases ,DNA Repair ,Duocarmycins ,Endodeoxyribonucleases ,Escherichia coli ,Escherichia coli Proteins ,Humans ,Indoles ,Mammals ,Mutation ,N-Glycosyl Hydrolases ,Rats ,Recombinant Proteins ,Sequence Analysis ,DNA ,3-methyladenine DNA glycosylase ,adozelesin ,alkylating agent ,excision repair ,UvrABC nuclease ,Biochemistry & Molecular Biology ,Biochemistry and cell biology - Abstract
Adozelesin is a synthetic analog of the antitumor antibiotic CC-1065, which alkylates the N3 of adenine in the minor groove in a sequence-selective manner. Since the cytotoxic potency of a DNA alkylating agent can be modulated by DNA excision repair system, we investigated whether nucleotide excision repair (NER) and base excision repair (BER) enzymes are able to excise the bulky DNA adduct induced by adozelesin. The UvrABC nuclease and 3-methyladenine-DNA glycosylase, that exhibit a broad spectrum of substrate specificity, were selected as typical NER and BER enzymes, respectively. The adozelesin-DNA adduct was first formed in the radiolabeled restriction DNA fragment and its excision by purified repair enzymes was monitored on a DNA sequencing gel. The treatment of the DNA adduct with a purified UvrABC nuclease and sequencing gel analysis of cleaved DNA showed that UvrABC nuclease was able to incise the adozelesin adduct. The incision site corresponded to the general nuclease incision site. Excision of this adduct by 3-methyladenine-DNA glycosylases was determined following the treatment of the DNA adduct with a homogeneous recombinant bacterial, rat and human 3-methyladenine-DNA glycosylases. Abasic sites generated by DNA glycosyalses were cleaved by the associated lyase activity of the E. coli formamidopyrimidine-DNA glycosylase (Fpg). Resolution of cleaved DNA on a sequencing gel showed that the DNA glycosylase from different sources could not release the N3-adenine adducts. A cytotoxicity assay using E. coli repair mutant strains showed that E. coli mutant strains defective in the uvrA gene were more sensitive to cell killing by adozelesin than E. coli mutant strain defective in the alkA gene or the wild type. These results suggest that the NER pathway seems to be the major excision repair system in protecting cells from the cytotoxicity of adozelesin.
- Published
- 2001
10. Antitumor activity of the investigational B7-H3 antibody-drug conjugate, vobramitamab duocarmazine, in preclinical models of neuroblastoma.
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Brignole C, Calarco E, Bensa V, Giusto E, Perri P, Ciampi E, Corrias MV, Astigiano S, Cilli M, Loo D, Bonvini E, Pastorino F, and Ponzoni M
- Subjects
- Child, Humans, Mice, Animals, Duocarmycins, B7 Antigens metabolism, Antibodies, Monoclonal, Humanized, Immunoconjugates pharmacology, Immunoconjugates therapeutic use, Neuroblastoma, Antineoplastic Agents pharmacology, Antineoplastic Agents therapeutic use
- Abstract
Introduction: B7-H3 is a potential target for pediatric cancers, including neuroblastoma (NB). Vobramitamab duocarmazine (also referred to as MGC018 and herein referred to as vobra duo) is an investigational duocarmycin-based antibody-drug conjugate (ADC) directed against the B7-H3 antigen. It is composed of an anti-B7-H3 humanized IgG1/kappa monoclonal antibody chemically conjugated through a cleavable valine-citrulline linker to a du ocarmycin-hydroxy b enzamide a zaindole (vc-seco-DUBA). Vobra duo has shown preliminary clinical activity in B7-H3-expressing tumors., Methods: B7-H3 expression was evaluated by flow-cytometry in a panel of human NB cell lines. Cytotoxicity was evaluated in monolayer and in multicellular tumor spheroid (MCTS) models by the water-soluble tetrazolium salt,MTS, proliferation assay and Cell Titer Glo 3D cell viability assay, respectively. Apoptotic cell death was investigated by annexin V staining. Orthotopic, pseudometastatic, and resected mouse NB models were developed to mimic disease conditions related to primary tumor growth, metastases, and circulating tumor cells with minimal residual disease, respectively., Results: All human NB cell lines expressed cell surface B7-H3 in a unimodal fashion. Vobra duo was cytotoxic in a dose-dependent and time-dependent manner against all cell lines (IC50 range 5.1-53.9 ng/mL) and NB MCTS (IC50 range 17.8-364 ng/mL). Vobra duo was inactive against a murine NB cell line (NX-S2) that did not express human B7-H3; however, NX-S2 cells were killed in the presence of vobra duo when co-cultured with human B7-H3-expressing cells, demonstrating bystander activity. In orthotopic and pseudometastatic mouse models, weekly intravenous treatments with 1 mg/kg vobra duo for 3 weeks delayed tumor growth compared with animals treated with an irrelevant (anti-CD20) duocarmycin-ADC. Vobra duo treatment for 4 weeks further increased survival in both orthotopic and resected NB models. Vobra duo compared favorably to TOpotecan-TEMozolomide (TOTEM), the standard-of-care therapy for NB relapsed disease, with tumor relapse delayed or arrested by two or three repeated 4-week vobra duo treatments, respectively. Further increased survival was observed in mice treated with vobra duo in combination with TOTEM. Vobra duo treatment was not associated with body weight loss, hematological toxicity, or clinical chemistry abnormalities., Conclusion: Vobra duo exerts relevant antitumor activity in preclinical B7-H3-expressing NB models and represents a potential candidate for clinical translation., Competing Interests: Competing interests: EB and DL are employees of MacroGenics and receive salary and stocks as part of their compensations. The other authors do not have competing interests to declare., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)
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- 2023
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11. Synthetic Studies on Complex Natural Products Based on Development of a Novel Synthetic Method for Heteroaromatic Skeleton
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Juri, Sakata
- Subjects
Pharmacology ,Biological Products ,Duocarmycins ,Indoles ,Cyclization ,Oximes ,Chemistry, Organic ,Pharmaceutical Science ,Quinolones ,Pyrroloiminoquinones - Abstract
Among my recent work on the syntheses of complex natural products based on the development of a novel synthetic method for the heteroaromatic skeleton, this article primarily deals with the total syntheses of (+)-CC-1065, isobatzeline A/B, and batzeline A. These syntheses were accomplished via a novel indole synthesis utilizing a ring expansion reaction of benzocyclobutenone oxime sulfonate as the key step. The 1,2-dihydro-3H-pyrrolo[3,2-e]indole segments of (+)-CC-1065 were rapidly constructed via a two-directional double-ring expansion strategy. Highly substituted pyrrolidine-fused common 5-chloro-2-methylthioindoles of isobatzeline A/B and batzeline A were constructed using a ring expansion reaction of benzocyclobutenone oxime sulfonate with NaSMe and a benzyne-mediated cyclization/functionalization reaction.
- Published
- 2022
12. Targeted clearance of senescent cells using an antibody-drug conjugate against a specific membrane marker
- Author
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Akang Leonard Bassey, Ana Sousa Manso, Mohammad Althubiti, Mark Frigerio, Andrew F. Kyle, Victoria M. Smith, Marta Poblocka, Marta Falcicchio, Ruth C. Barber, XiaoBo Sheng, Salvador Macip, Universitat Oberta de Catalunya (UOC), University of Leicester, and Babraham Research Campus
- Subjects
Senescence ,Antibody-drug conjugate ,Immunoconjugates ,Science ,Biology ,Medical sciences ,Epitope ,Article ,Cell Line ,Duocarmycins ,In vivo ,Fibrosis ,Senotherapeutics ,target identification ,Target identification ,medicine ,marcador biológico ,Humans ,identificació d'objectius ,marcador biològic ,Senolytic ,Cellular Senescence ,Ciències de la salut ,identificación de objetivos ,Multidisciplinary ,medicine.disease ,biological marker ,Isotype ,senescence cells ,Ciencias de la salud ,Gene Expression Regulation ,cèl·lules senescents ,biology.protein ,Cancer research ,Medicine ,Antibody ,Tumor Suppressor Protein p53 ,células senescentes ,beta 2-Microglobulin - Abstract
A wide range of diseases have been shown to be influenced by the accumulation of senescent cells, from fibrosis to diabetes, cancer, Alzheimer’s and other age-related pathologies. Consistent with this, clearance of senescent cells can prolong healthspan and lifespan in in vivo models. This provided a rationale for developing a new class of drugs, called senolytics, designed to selectively eliminate senescent cells in human tissues. The senolytics tested so far lack specificity and have significant off-target effects, suggesting that a targeted approach could be more clinically relevant. Here, we propose to use an extracellular epitope of B2M, a recently identified membrane marker of senescence, as a target for the specific delivery of toxic drugs into senescent cells. We show that an antibody–drug conjugate (ADC) against B2M clears senescent cells by releasing duocarmycin into them, while an isotype control ADC was not toxic for these cells. This effect was dependent on p53 expression and therefore more evident in stress-induced senescence. Non-senescent cells were not affected by either antibody, confirming the specificity of the treatment. Our results provide a proof-of-principle assessment of a novel approach for the specific elimination of senescent cells using a second generation targeted senolytic against proteins of their surfaceome, which could have clinical applications in pathological ageing and associated diseases.
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- 2021
13. Duocarmycin-based antibody–drug conjugates as an emerging biotherapeutic entity for targeted cancer therapy: Pharmaceutical strategy and clinical progress
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Ming-Hai Wang, Hui Zhao, Xiang-Min Tong, Hangping Yao, and Rachel Hudson
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Pharmacology ,Drug ,Antibody-drug conjugate ,Immunoconjugates ,business.industry ,media_common.quotation_subject ,Drug Evaluation, Preclinical ,Cancer therapy ,Antineoplastic Agents ,Computational biology ,body regions ,Duocarmycins ,chemistry.chemical_compound ,Drug Development ,chemistry ,Neoplasms ,Drug Discovery ,Animals ,Humans ,Medicine ,business ,Duocarmycin ,media_common - Abstract
Duocarmycins are a class of DNA minor-groove-binding alkylating molecules. For the past decade, various duocarmycin analogues have been used as payloads in the development of antibody-drug conjugates (ADCs). Currently, more than 15 duocarmycin-based ADCs have been studied preclinically, and some of them such as SYD985 have been granted Fast-Track Designation status. Nevertheless, progress in duocarmycin-based ADCs also faces challenges, with setbacks including the termination of BMS-936561/MDX-1203. In this review, we discuss issues associated with the efficacy, pharmacokinetic profile, and toxicological activity of these biotherapeutics. Furthermore, we summarize the latest advances in duocarmycin-based ADCs that have different target specificities and linker chemistries. Evidence from preclinical and clinical studies has indicated that duocarmycin-based ADCs are promising biotherapeutics for oncological application in the future.
- Published
- 2021
14. Rapid Construction of a Chloromethyl-Substituted Duocarmycin-like Prodrug.
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Bengtsson C and Gravenfors Y
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- Duocarmycins, Amination, Prodrugs
- Abstract
The construction of duocarmycin-like compounds is often associated with lengthy synthetic routes. Presented herein is the development of a short and convenient synthesis of a type of duocarmycin prodrug. The 1,2,3,6-tetrahydropyrrolo[3,2- e ]indole-containing core is here constructed from commercially available Boc-5-bromoindole in four steps and 23% overall yield, utilizing a Buchwald-Hartwig amination followed by a sodium hydride-induced regioselective bromination. In addition, protocols for selective mono- and di-halogenations of positions 3 and 4 were also developed, which could be useful for further exploration of this scaffold.
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- 2023
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15. Design, synthesis, nuclear localization, and biological activity of a fluorescent duocarmycin analog, HxTfA.
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Kiakos, Kostantinos, Englinger, Bernhard, Yanow, Stephanie K., Wernitznig, Debora, Jakupec, Michael A., Berger, Walter, Keppler, Bernhard K., Hartley, John A., Lee, Moses, and Patil, Pravin C.
- Subjects
- *
BIOACTIVE compounds , *ANTIMALARIALS , *ANTINEOPLASTIC agents , *DRUG development , *DRUG design , *DRUG synthesis - Abstract
HxTfA 4 is a fluorescent analog of a potent cytotoxic and antimalarial agent, TfA 3 , which is currently being investigated for the development of an antimalarial vaccine, PlasProtect®. HxTfA contains a p-anisylbenzimidazole or Hx moiety, which is endowed with a blue emission upon excitation at 318 nm; thus enabling it to be used as a surrogate for probing the cellular fate of TfA using confocal microscopy, and addressing the question of nuclear localization. HxTfA exhibits similar selectivity to TfA for A-tract sequences of DNA, alkylating adenine-N3, albeit at 10-fold higher concentrations. It also possesses in vitro cytotoxicity against A549 human lung carcinoma cells and Plasmodium falciparum. Confocal microscopy studies showed for the first time that HxTfA, and by inference TfA, entered A549 cells and localized in the nucleus to exert its biological activity. At biologically relevant concentrations, HxTfA elicits DNA damage response as evidenced by a marked increase in the levels of γH2AX observed by confocal microscopy and immunoblotting studies, and ultimately induces apoptosis. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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16. How can the potential of the duocarmycins be unlocked for cancer therapy?
- Author
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Goreti Ribeiro Morais, Zoë Jukes, Paul M. Loadman, and Klaus Pors
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0301 basic medicine ,Drug ,media_common.quotation_subject ,Cancer therapy ,Computational biology ,Antibodies ,Duocarmycins ,Structure-Activity Relationship ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Neoplasms ,Drug Discovery ,Animals ,Humans ,Medicine ,Prodrugs ,Antineoplastic Agents, Alkylating ,Duocarmycin ,media_common ,Pharmacology ,business.industry ,Prodrug ,Drug Resistance, Multiple ,030104 developmental biology ,chemistry ,Drug Resistance, Neoplasm ,030220 oncology & carcinogenesis ,business - Abstract
The duocarmycins belong to a class of agent that has fascinated scientists for over four decades. Their exquisite potency, unique mechanism of action, and efficacy in multidrug-resistant tumour models makes them attractive to medicinal chemists and drug hunters. However, despite great advances in fine-tuning biological activity through structure-activity relationship studies (SARS), no duocarmycin-based therapeutic has reached clinical approval. In this review, we provide an overview of the most promising strategies currently used and include both tumour-targeted prodrug approaches and antibody-directed technologies.
- Published
- 2021
17. A Peptide–Duocarmycin Conjugate Targeting the Thomsen-Friedenreich Antigen Has Potent and Selective Antitumor Activity
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Mark Searcey, David Russell, Marco M. D. Cominetti, Oliver Charles Cartwright, and Andrew M. Beekman
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Protein subunit ,Biomedical Engineering ,Pharmaceutical Science ,Antineoplastic Agents ,Bioengineering ,Peptide ,02 engineering and technology ,01 natural sciences ,Cathepsin B ,Duocarmycins ,chemistry.chemical_compound ,Antigen ,Cell Line, Tumor ,Humans ,Antigens, Tumor-Associated, Carbohydrate ,Amino Acid Sequence ,Peptide sequence ,Duocarmycin ,Cell Proliferation ,Pharmacology ,chemistry.chemical_classification ,Thomsen-Friedenreich Antigen ,010405 organic chemistry ,Organic Chemistry ,021001 nanoscience & nanotechnology ,Molecular biology ,0104 chemical sciences ,chemistry ,Peptides ,0210 nano-technology ,Biotechnology ,Conjugate - Abstract
Solid phase synthesis allowed the rapid generation of a peptide-drug conjugate. A peptide targeting the Thomsen-Friedenreich antigen (TFα) was conjugated to the alkylating subunit of the potent cytotoxin duocarmycin SA. The compound, containing a cathepsin B cleavable linker, was shown to be active and selective against TFα expressing tumour cell lines.
- Published
- 2020
18. Structural evolution of a DNA repair self-resistance mechanism targeting genotoxic secondary metabolites
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Dale L. Boger, Elwood A. Mullins, Jonathan Dorival, Gong-Li Tang, and Brandt F. Eichman
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DNA Repair ,Science ,General Physics and Astronomy ,Streptomyces ,Article ,General Biochemistry, Genetics and Molecular Biology ,DNA Glycosylases ,Duocarmycins ,chemistry.chemical_compound ,Bacterial Proteins ,Biosynthesis ,Gene cluster ,Gene ,X-ray crystallography ,chemistry.chemical_classification ,Natural products ,Multidisciplinary ,biology ,Chemistry ,DNA damage and repair ,General Chemistry ,biology.organism_classification ,In vitro ,Anti-Bacterial Agents ,Enzyme ,DNA repair enzymes ,Biochemistry ,DNA glycosylase ,Multigene Family ,DNA ,DNA Damage ,Mutagens - Abstract
Microbes produce a broad spectrum of antibiotic natural products, including many DNA-damaging genotoxins. Among the most potent of these are DNA alkylating agents in the spirocyclopropylcyclohexadienone (SCPCHD) family, which includes the duocarmycins, CC-1065, gilvusmycin, and yatakemycin. The yatakemycin biosynthesis cluster in Streptomyces sp. TP-A0356 contains an AlkD-related DNA glycosylase, YtkR2, that serves as a self-resistance mechanism against yatakemycin toxicity. We previously reported that AlkD, which is not present in an SCPCHD producer, provides only limited resistance against yatakemycin. We now show that YtkR2 and C10R5, a previously uncharacterized homolog found in the CC-1065 biosynthetic gene cluster of Streptomyces zelensis, confer far greater resistance against their respective SCPCHD natural products. We identify a structural basis for substrate specificity across gene clusters and show a correlation between in vivo resistance and in vitro enzymatic activity indicating that reduced product affinity—not enhanced substrate recognition—is the evolutionary outcome of selective pressure to provide self-resistance against yatakemycin and CC-1065., Microbial DNA glycosylases associated with the biosynthesis of DNA-damaging antibiotics have evolved self-resistance for their cognate natural products. Here, the authors provide evidence that cellular self-resistance is enabled by reduced affinity of the glycosylases for the excision products of the corresponding DNA lesions.
- Published
- 2021
19. Cytochrome P450 Binding and Bioactivation of Tumor-Targeted Duocarmycin Agents
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Goreti Ribeiro Morais, Emily E. Scott, Aaron G. Bart, Klaus Pors, Venu R. Vangala, and Paul M. Loadman
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Stereochemistry ,Metabolite ,Pharmaceutical Science ,chemistry.chemical_compound ,Duocarmycins ,Cytochrome P-450 Enzyme System ,Cell Line, Tumor ,Cytochrome P-450 CYP1A1 ,Humans ,Prodrugs ,Benzofuran ,Cytochrome P450 Family 2 ,Duocarmycin ,Pharmacology ,Indole test ,Natural product ,biology ,Cytochrome P450 ,Stereoisomerism ,Articles ,Prodrug ,Antineoplastic Agents, Phytogenic ,chemistry ,Drug Design ,biology.protein ,Drug metabolism ,Biomarkers - Abstract
Duocarmycin natural products are promising anticancer cytotoxins but too potent for systemic use. Re-engineering of the duocarmycin scaffold has enabled the discovery of prodrugs designed for bioactivation by tissue-specific cytochrome P450 (P450) enzymes. Lead prodrugs bioactivated by both P450 isoforms CYP1A1 and CYP2W1 have shown promising results in xenograft studies; however, to fully understand the potential of these agents it is desirable to compare dual-targeting compounds with isoform-selective analogs. Such redesign requires insight into the molecular interactions with these P450 enzymes. Herein binding and metabolism of the individual stereoisomers of the indole-based duocarmycin prodrug ICT2700 and a nontoxic benzofuran analog ICT2726 were evaluated with CYP1A1 and CYP2W1, revealing differences exploitable for drug design. Although enantiomers of both compounds bound to and were metabolized by CYP1A1, the stereochemistry of the chloromethyl fragment was critical for CYP2W1 interactions. CYP2W1 differentially binds the S enantiomer of ICT2726, and its metabolite profile could potentially be used as a biomarker to identify CYP2W1 functional activity. In contrast to benzofuran-based ICT2726, CYP2W1 differentially binds the R isomer of the indole-based ICT2700 over the S stereoisomer. Thus the ICT2700 R configuration warrants further investigation as a scaffold to favor CYP2W1-selective bioactivation. Furthermore, structures of both duocarmycin S enantiomers with CYP1A1 reveal orientations correlating with nontoxic metabolites, and further drug design optimization could lead to a decrease of CYP1A1 bioactivation. Overall, distinctive structural features present in the two P450 active sites can be useful for improving P450—and thus tissue-selective—bioactivation. SIGNIFICANCE STATEMENT: Prodrug versions of the natural product duocarmycin can be metabolized by human tissue-specific cytochrome P450 (P450) enzymes 1A1 and 2W1 to form an ultrapotent cytotoxin and/or high affinity 2W1 substrates to potentially probe functional activity in situ. The current work defines the binding and metabolism by both P450 enzymes to support the design of duocarmycins selectively activated by only one human P450 enzyme.
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- 2021
20. The CYP2W1 enzyme: regulation, properties and activation of prodrugs.
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Guo, Jia, Johansson, Inger, Mkrtchian, Souren, and Ingelman-Sundberg, Magnus
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- *
CYTOCHROMES , *INDOLINE , *PRODRUGS , *EPIGENESIS , *COLON cancer , *PROTEIN expression , *METASTASIS - Abstract
CYP2W1 is expressed in the course of development of the gastrointestinal tract, silenced after birth in intestine and colon by epigenetic modifications, but activated following demethylation in colorectal cancer (CRC). The expression levels in CRC positively correlate with the degree of malignancy, are higher in metastases and are predictive of colon cancer survival. The CYP2W1 transcripts have been detected also in hepatocellular carcinoma, adrenocortical carcinoma, childhood rhabdomyosarcoma and breast cancer; however, here the protein expression remains to be confirmed. The CYP2W1 enzyme has an inverted orientation in the endoplasmic reticulum membrane, as compared to other cytochrome P450s and its immediate electron donor is unknown. Several lipid ligands have been proposed as endogenous substrates, among which retinol derivatives appear to have the highest affinities. However, the role of CYP2W1 in the endogenous and tumor localized metabolism of retinol derivatives has yet to be clarified. Indolines constitute high affinity exogenous compounds and specific chloromethylindolines have been shown to be activated by CYP2W1 into cytotoxic productsin vitroand alsoin vivo, inhibiting the growth of human colon tumors in a mouse xenograft model. The CRC specific localization of CYP2W1 and its effective prodrug activation makes it a very promising target for future development of cancer therapeutics. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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21. A non-reactive natural product precursor of the duocarmycin family has potent and selective antimalarial activity
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Gerard D. Wright, Michael A. Cook, Sarah Lemcke, Arne Alder, Wenliang Wang, Min Xu, Nicole S. Struck, Jarrod W. Johnson, Janis Rambow, Daniel Pallant, and Tim W. Gilberger
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Cell division ,Clinical Biochemistry ,Biology ,Biochemistry ,Schizogony ,chemistry.chemical_compound ,Antimalarials ,Duocarmycins ,Drug Discovery ,Animals ,Humans ,Parasites ,Molecular Biology ,Duocarmycin ,Pharmacology ,Biological Products ,Natural product ,DNA replication ,Plasmodium falciparum ,DNA ,biology.organism_classification ,Malaria ,chemistry ,Eukaryotic chromosome fine structure ,Molecular Medicine ,Folic Acid Antagonists - Abstract
Summary We identify a selective nanomolar inhibitor of blood-stage malarial proliferation from a screen of microbial natural product extracts. The responsible compound, PDE-I2, is a precursor of the anticancer duocarmycin family that preserves the class's sequence-specific DNA binding but lacks its signature DNA alkylating cyclopropyl warhead. While less active than duocarmycin, PDE-I2 retains comparable antimalarial potency to chloroquine. Importantly, PDE-I2 is >1,000-fold less toxic to human cell lines than duocarmycin, with mitigated impacts on eukaryotic chromosome stability. PDE-I2 treatment induces severe defects in parasite nuclear segregation leading to impaired daughter cell formation during schizogony. Time-of-addition studies implicate parasite DNA metabolism as the target of PDE-I2, with defects observed in DNA replication and chromosome integrity. We find the effect of duocarmycin and PDE-I2 on parasites is phenotypically indistinguishable, indicating that the DNA binding specificity of duocarmycins is sufficient and the genotoxic cyclopropyl warhead is dispensable for the parasite-specific selectivity of this compound class.
- Published
- 2021
22. Unexpected Syntheses of seco-Cyclopropyltetrahydroquinolines >From a Radical 5-Exo-Trig Cyclization Reaction: Analogs of CC-1065 and the Duocarmycins
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Moses Lee, LuAnne McNulty, Brian Lingerfelt, Heather Townes, Lori Forrest, and Hari Pati
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seco-Cyclopropyltetrahydroquinoline ,CC-1065 ,duocarmycins ,(chloromethyl)furano[e]indoline ,(chloromethyl)furano[f]tetrahydroquinoline. ,Organic chemistry ,QD241-441 - Abstract
Analogs of the seco-cyclopyrroloindoline (seco-CPI), the DNA alkylation pharmacophore of CC-1065 and the duocarmycins, can be prepared through a 5-exo-trig radical cyclization of a free radical and a 3-chloro-2-allylic moiety. This manuscript reports an unexpected discovery that, depending on the structure and stability of the free radical, the cyclization process leads to the production of an appreciable amount of seco- cyclopropyltetrahydroquinolines 7a-d along with the seco-cyclopropoyltetra- hydroindoline products (6a-e). For instance, free radical reaction of the bromoallylic chloride 5a produced an equal amount of 6-benzyloxy-N-t-butoxycarbonyl-3- (chloromethyl)furano[e]indoline (6a), and 7-benzyloxy-N-t-butoxycarbonyl-3-chloro- 1,2,3,4-tetrahydrofurano[f]quinoline (7a). Three other examples that produced mixtures of indoline and quinoline products are provided. In only one of the examples reported in this manuscript, the 6-benzyloxy-N-t-butoxycarbonyl-3-(chloromethyl)benzo[e]indoline, was a seco-CBI precursor 6e formed exclusively, consistent with literature precedents.
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- 2004
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23. A Novel Depurination Methodology to Assess DNA Alkylation of Chloro-Bis-Seco-Cyclopropylbenzoindoles Allowed for Comparison of Minor-Groove Reactivity
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Donglu Zhang, Binqing Wei, Jun Guo, Thomas H. Pillow, Buyun Chen, Chenghong Zhang, Leanna Staben, Dewakar Sangaraju, Gail Lewis Phillips, Dian Su, Yong Ma, Cyrus Khojasteh, Sudheer Bobba, Jianshuang Wang, Peter S. Dragovich, and Shuai Wang
- Subjects
Alkylating Agents ,Alkylation ,Stereochemistry ,Population ,Pharmaceutical Science ,030226 pharmacology & pharmacy ,Adduct ,DNA Adducts ,Duocarmycins ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,DNA Crosslinking ,DNA adduct ,Reactivity (chemistry) ,education ,Antineoplastic Agents, Alkylating ,Pharmacology ,education.field_of_study ,Adenine ,DNA ,DNA Alkylation ,chemistry ,030220 oncology & carcinogenesis ,Depurination - Abstract
Duocarmycins [including cyclopropyl pyrroloindole (CPI) or cyclopropyl benzoindole (CBI)] are a class of DNA minor-groove alkylators and seco-CPI/CBIs are synthetic pro-forms that can spirocyclize to CPI/CBI. Bis-CPI/CBIs are potential drug candidates because of their enhanced cytotoxicity from DNA crosslinking, but it is difficult to analyze them for structure-activity correlation because of their DNA reactivity. To study their DNA alkylation, neutral thermal hydrolysis has been frequently applied to process depurination. However, unwanted side reactions under this condition have been reported, which could lead to poor correlation of DNA alkylation data with efficacy results, especially for bis-CPI/CBIs. In this study, an acidic depurination method was developed and applied for analysis of DNA alkylation and shown to be an easier and milder method than the traditional neutral thermal hydrolysis. DNA alkylation and stability of three bis-seco-CBIs were characterized in comparison with two mono-seco-CPIs. The results suggested that: 1) The acidic depurination method was capable of capturing a more representative population, sometimes a different population, of DNA adducts as they existed on DNA compared with the heat depurination method. 2) Di-adenine adducts were captured as expected for the CBI dimers, although the major type of adduct was still mono-adenine adducts. 3) The rate of DNA alkylation, DNA adduct profile, and relative amounts of di-adduct versus mono-adduct were significantly affected by the size, and possibly lipophilicity, of the nonalkylating part of the molecules. 4) Spirocyclization and amide hydrolysis represented two major pathways of degradation. Overall, by applying acidic depurination analyses, this study has illustrated DNA adduct characteristics of novel bis-seco-CBIs with dominating mono-alkylation and provides an alternative method for evaluating DNA minor-groove alkylators. These findings provide an effective analytical tool to evaluate DNA alkylators and to study the DNA alkylation that is a disposition mechanism of these compounds.
- Published
- 2019
24. Characterizing and understanding the formation of cysteine conjugates and other by-products in a random, lysine-linked antibody drug conjugate
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Michael Hay, Scott A. Miller, Naresh Chennamsetty, Difei Qiu, and Yande Huang
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Antibody-drug conjugate ,Immunoconjugates ,Stereochemistry ,Immunology ,Lysine ,Conjugated system ,Amidine ,Duocarmycins ,chemistry.chemical_compound ,Antibody drug conjugate ,Report ,Drug Discovery ,Humans ,Immunology and Allergy ,Moiety ,Cysteine ,Duocarmycin ,mass spectrometry ,lysine-linked conjugation ,chemistry ,Immunoglobulin G ,cysteine-linked conjugation ,Reports ,Conjugate - Abstract
This study describes the characterization of conjugation sites for a random, lysine conjugated 2-iminothiolane (2-IT) based antibody-drug-conjugate synthesized from an IgG1 antibody and a duocarmycin analog-based payload-linker. Of the 80 putative lysine sites, 78 were found to be conjugated via tryptic peptide mapping and LC-HRMS. Surprisingly, seven cysteine-linked conjugated peptides were also detected resulting from the conjugation of cysteine residues derived from the four inter-chain disulfide bonds during the reaction. This unexpected finding could be attributed to the free thiols of the 2-IT thiolated antibody intermediates and/or the 4-mercaptobutanamide by-product resulting from the hydrolysis of 2-IT. These free thiols could cause the four inter-chain disulfide bonds of the antibody to scramble via intra- or inter-molecular attack. The presence of only pair of non-reactive (unconjugated) lysine residues, along with the four intact intra-chain disulfide bonds, is attributed to their poor accessibility, which is consistent with solvent accessibility modeling analysis. We also discovered a major by-product derived from the hydrolysis of the amidine moiety of the N-terminus conjugate. In contrast, the amidine moiety in lysine-linked conjugates appeared stable. Based on our results, we propose plausible formation mechanisms of cysteine-linked conjugates and the hydrolysis of the N-terminus conjugate, which provide scientific insights that are beneficial to process development and drug quality control.
- Published
- 2021
25. A Platform for the Generation of Site-Specific Antibody-Drug Conjugates That Allows for Selective Reduction of Engineered Cysteines
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Bas P. A. Kokke, C. Marco Timmers, Henri Johannes Spijker, Jan Schouten, Pascal Renart Verkerk, Wim H. A. Dokter, Ruud Coumans, Patrick G. Groothuis, Ruud Ubink, Patrick Henry Beusker, Miranda M. C. van der Lee, Gerry J. A. Ariaans, and Marion Blomenrohr
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Drug ,Models, Molecular ,Immunoconjugates ,media_common.quotation_subject ,Biomedical Engineering ,Pharmaceutical Science ,Bioengineering ,02 engineering and technology ,Computational biology ,01 natural sciences ,Duocarmycins ,Mice ,Therapeutic index ,Cell Line, Tumor ,Neoplasms ,Animals ,Humans ,Cysteine ,media_common ,Pharmacology ,Antibiotics, Antineoplastic ,biology ,010405 organic chemistry ,Chemistry ,Organic Chemistry ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,body regions ,Specific antibody ,ComputingMethodologies_PATTERNRECOGNITION ,Immunoglobulin G ,biology.protein ,Antibody ,0210 nano-technology ,Hydrophobic and Hydrophilic Interactions ,Oxidation-Reduction ,Biotechnology ,Conjugate - Abstract
Engineering cysteines at specific sites in antibodies to create well-defined ADCs for the treatment of cancer is a promising approach to increase the therapeutic index and helps to streamline the manufacturing process. Here, we report the development of an
- Published
- 2020
26. Galactose‐modified duocarmycin prodrugs as senolytics
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Juan Pedro Martinez-Barbera, Lutz F. Tietze, Anthony G. Uren, Ana Guerrero, Nicolás Herranz, Romain Guiho, Dominic J. Withers, Jesús Gil, Wellcome Trust, and Medical Research Council
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0301 basic medicine ,Aging ,Geriatrics & Gerontology ,CLEARANCE ,senescence ,Apoptosis ,Craniopharyngioma ,Mice ,chemistry.chemical_compound ,0302 clinical medicine ,PROGRAM ,Bystander effect ,senescence-associated β-galactosidase ,Cytotoxic T cell ,Prodrugs ,11 Medical and Health Sciences ,Cellular Senescence ,Tissue homeostasis ,0303 health sciences ,Antibiotics, Antineoplastic ,CHEMOTHERAPY ,Prodrug ,3. Good health ,Cell biology ,030220 oncology & carcinogenesis ,senescence‐associated β‐galactosidase ,prodrug ,Life Sciences & Biomedicine ,medicine.drug ,Senescence ,duocarmycin ,Senescence-associated beta-galactosidase ,Biology ,CONTRIBUTES ,Cell Line ,GLYCOSIDIC PRODRUGS ,Duocarmycins ,03 medical and health sciences ,medicine ,senescence-associated beta-galactosidase ,Animals ,Humans ,Doxorubicin ,Duocarmycin ,030304 developmental biology ,Original Paper ,Science & Technology ,Galactose ,Neoplasms, Experimental ,Cell Biology ,06 Biological Sciences ,beta-Galactosidase ,Coculture Techniques ,Mice, Inbred C57BL ,SENESCENT CELLS ,030104 developmental biology ,chemistry ,senolytics ,SECRETOME ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Senescence is a stable growth arrest that impairs the replication of damaged, old or preneoplastic cells, therefore contributing to tissue homeostasis. Senescent cells accumulate during ageing and are associated with cancer, fibrosis and many age‐related pathologies. Recent evidence suggests that the selective elimination of senescent cells can be effective on the treatment of many of these senescence‐associated diseases. A universal characteristic of senescent cells is that they display elevated activity of the lysosomal β‐galactosidase, and this has been exploited as a marker for senescence (senescence‐associated β‐galactosidase activity). Consequently, we hypothesized that galactose‐modified cytotoxic prodrugs will be preferentially processed by senescent cells, resulting in their selective killing. Here, we show that different galactose‐modified duocarmycin (GMD) derivatives preferentially kill senescent cells. GMD prodrugs induce selective apoptosis of senescent cells in a lysosomal β‐galactosidase (GLB1)‐dependent manner. GMD prodrugs can eliminate a broad range of senescent cells in culture, and treatment with a GMD prodrug enhances the elimination of bystander senescent cells that accumulate upon whole‐body irradiation treatment of mice. Moreover, taking advantage of a mouse model of adamantinomatous craniopharyngioma (ACP), we show that treatment with a GMD prodrug selectively reduced the number of β‐catenin‐positive preneoplastic senescent cells. In summary, the above results make a case for testing the potential of galactose‐modified duocarmycin prodrugs to treat senescence‐related pathologies., Galactose‐modified duocarmycin prodrugs get preferentially processed in senescent cells and behave as senolytics.
- Published
- 2020
27. Therapeutic efficacy of a novel humanized antibody-drug conjugate recognizing plexin-semaphorin-integrin domain in the RON receptor for targeted cancer therapy
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Tian Hao Weng, Zhi Gang Wu, Yi Zhi Liu, Xiang Min Tong, Liang Feng, Ming Hai Wang, Hangping Yao, Sreedhar Reddy Suthe, and Chen Yu Hu
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0301 basic medicine ,Cancer Research ,Immunoconjugates ,PSI domain ,Receptor internalization ,Humanized antibody ,Mice ,chemistry.chemical_compound ,0302 clinical medicine ,Neoplasms ,Antineoplastic Combined Chemotherapy Protocols ,Immunology and Allergy ,Antibody-drug conjugates ,Internalization ,media_common ,biology ,Epithelial cancer, cancer stem cells ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Humanization ,Oncology ,Monomethyl auristatin E ,030220 oncology & carcinogenesis ,Drug delivery ,Molecular Medicine ,Female ,RON receptor tyrosine kinase ,Oligopeptides ,Research Article ,Monoclonal antibody ,Maximum Tolerated Dose ,medicine.drug_class ,media_common.quotation_subject ,Immunology ,Antibodies, Monoclonal, Humanized ,lcsh:RC254-282 ,Duocarmycins ,03 medical and health sciences ,Protein Domains ,In vivo ,Cancer stem cell ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Pharmacology ,CD44 ,Receptor Protein-Tyrosine Kinases ,Tumor xenograft model ,Therapeutic efficacy ,Xenograft Model Antitumor Assays ,body regions ,030104 developmental biology ,chemistry ,Cancer research ,biology.protein - Abstract
Background Antibody-drug conjugates (ADCs) targeting the RON receptor, a tumorigenic factor contributing to cancer malignancy, has been considered as a novel strategy for cancer therapy. Here we describe a humanized antibody recognizing the RON plexin-semaphorin-integrin (PSI) domain with increased drug delivery capability for potential clinical application. Method Monoclonal antibody PCM5B14 specific to the human and monkey RON PSI domain was generated and characterized by various immunological methods. Humanized antibody H5B14 was created by grafting PCM5B14 complementarity-determining regions into human IgG1/κ acceptor frameworks and conjugated with monomethyl auristatin E and duocarmycin to form two H5B14-based ADCs. Stability of H5B14-based ADCs in human plasma was measured using hydrophobic interaction chromatography. Various biochemical and biological assays were used to determine ADC- regulated RON internalization, cell viability, spheroid formation, and death of cancer stem-like cells. Efficacies of H5B14-based ADCs in vivo were validated using tumor xenograft models. Maximal tolerated doses of H5B14-based ADCs were established in mice. Results H5B14 was highly specific to the human RON PSI domain and superior over other anti-RON ADCs in induction of RON internalization in various cancer cell lines tested. H5B14-based ADCS had a drug to antibody ratio of ~ 3.70:1 and were stable in human plasma with a minimal dissociation within a 10-day period. Functionally, H5B14-mediated drug delivery decreased cell viability at early stages with an average IC50 at ~ 20 nM in multiple cancer cell lines examined. H5B14-based ADCs also inhibited spheroid formation and caused death of cancer stem-like cells with RON+/CD44+/ESA+ phenotypes. In vivo, H5B14-based ADCs in a single injection inhibited tumor xenograft growth mediated by multiple cancer cell lines. Tumoristatic concentrations calculated from xenograft tumor models were in the range of 0.63 to 2.0 mg/kg bodyweight. Significantly, H5B14-based ADCs were capable of eradicating tumors at variable levels across multiple xenograft models regardless their malignant statuses. Toxicologically, H5B14-based ADCs were well tolerated in mice up to 60 mg/kg. Conclusion H5B14-based ADCs targeting the RON PSI domain are superior in inducing RON internalization, leading to robust drug delivery and overall inhibition and eradication of tumors in multiple xenograft models. These findings warrant H5B14-based ADCs for clinical trials in the future.
- Published
- 2019
28. Promiximab-duocarmycin, a new CD56 antibody-drug conjugates, is highly efficacious in small cell lung cancer xenograft models
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Ruixue Wang, Yiran Tao, Shuli Yi, Yu Liu, Yuyin Fu, Ruirui Zhang, Ying Lu, Yuqin Yao, Jinliang Yang, Wenting Li, Lin Yu, Qinhuai Lai, Lantu Gou, Ligong Chen, and Yuxi Wang
- Subjects
0301 basic medicine ,medicine.medical_treatment ,Cell ,antibody-drug conjugates ,Spleen ,duocarmycins ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,In vivo ,medicine ,Duocarmycin ,biology ,Immunotherapy ,In vitro ,respiratory tract diseases ,030104 developmental biology ,medicine.anatomical_structure ,Oncology ,chemistry ,Cell culture ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,CD56 ,small cell lung cancer ,Antibody ,Research Paper - Abstract
Small cell lung cancer (SCLC) is of a highly invasive and metastatic lung cancer subtype and there had not been effective targeted therapies. CD56, a cell surface marker highly expressed on most SCLC, is a promising therapeutic target for treatment of this aggressive cancer. In this study, we generated a novel anti-CD56 antibody named promiximab, characterized by high affinity, internalization and tumor specificity. Then, the promiximab was conjugated with a potent DNA alkylating agent duocarmycin via reduced interchain disulfides to yield the promiximab-Duocarmycin (promiximab-DUBA) conjugates. Mass spectrometry analysis showed promiximab-DUBA had an average DAR (Drug-to-Antibody Ratio) of about 2.04. In vitro, promiximab-DUBA exerted strong inhibitory effects on SCLC cell lines NCI-H526, NCI-H524 and NCI-H69, with IC50 values of 0.07 nmol/L, 0.18 nmol/L and 0.29 nmol/L, respectively. In vivo antitumor activity, promiximab-DUBA at the dose of 5 mg/kg and 10 mg/kg every three days with a total of three times were sufficient to induce sustained regression of NCI-H526 tumors over control treatment with promiximab. Mostly, no recurrence was observed until 65 days post treatment with promiximab-DUBA. In the NCI-H69 subcutaneous xenograft model, significant inhibition of tumor growth was also observed following administration of promiximab-DUBA at the dose of 5 mg/kg or 10 mg/kg. Moreover, body weight and histopathology of major organs (liver, spleen, heart, lung and kidney) showed no significant changes after treatment of promiximab-DUBA. In conclusion, promiximab-DUBA is highly efficacious in small cell lung cancer xenograft models, and provides a new immunotherapy approach for SCLC.
- Published
- 2017
29. Probing cytochrome P450 (CYP) bioactivation with chloromethylindoline bioprecursors derived from the duocarmycin family of compounds
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Kersti Karu, Helen M. Sheldrake, Steve D. Shnyder, Natalia Ortuzar, Francis M. Barnieh, Laurence H. Patterson, Klaus Pors, Mark Searcey, Daniela Presa, Goreti Ribeiro Morais, and Paul M. Loadman
- Subjects
Indoles ,CYP1B1 ,Clinical Biochemistry ,Pharmaceutical Science ,Antineoplastic Agents ,Pharmacology ,Biochemistry ,Duocarmycins ,Structure-Activity Relationship ,chemistry.chemical_compound ,Cytochrome P-450 Enzyme System ,Cell Line, Tumor ,Drug Discovery ,Cytochrome P-450 Enzyme Inhibitors ,Humans ,Cytotoxicity ,Molecular Biology ,Duocarmycin ,Cell Proliferation ,Indole test ,Dose-Response Relationship, Drug ,Molecular Structure ,biology ,CYP3A4 ,Organic Chemistry ,CYP1A2 ,Cytochrome P450 ,chemistry ,Cancer cell ,biology.protein ,Molecular Medicine ,Drug Screening Assays, Antitumor - Abstract
The duocarmycins belong to a class of agent which has great potential for use in cancer therapy. Their exquisite potency means they are too toxic for systemic use, and targeted approaches are required to unlock their clinical potential. In this study, we have explored seco-OH-chloromethylindoline (CI) duocarmycin-based bioprecursors for their potential for cytochrome P450 (CYP)-mediated cancer cell kill. We report on synthetic and biological explorations of racemic seco-CI-MI, where MI is a 5-methoxy indole motif, and dehydroxylated analogues. We show up to a 10-fold bioactivation of de-OH CI-MI and a fluoro bioprecursor analogue in CYP1A1-transfected cells. Using CYP bactosomes, we also demonstrate that CYP1A2 but not CYP1B1 or CYP3A4 has propensity for potentiating these compounds, indicating preference for CYP1A bioactivation.
- Published
- 2021
30. Controlling the radical 5-exo-trig cyclization, and selective synthesis of seco-iso-cyclopropylfurano[e]indoline (seco-iso-CFI) and seco-cyclopropylthiophene[e]indoline (seco-CTI) DNA alkylating subunit of the duocarmycins.
- Author
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Patil, Pravin, Cousins, Karalyne, Smith, Mallory, Wieskamp, Sarah, Ferrara, Maddi, Bruce, Chrystal D., and Lee, Moses
- Subjects
- *
RING formation (Chemistry) , *CHEMICAL synthesis , *INDOLINE , *DNA alkylation , *TOLUENE , *BENZOFURAN - Abstract
Abstract: The free radical cyclization of benzofuran bromo-allylic chloride 6 in toluene produced an unpredicted mixture of 6-benzyloxy-N-t-Boc-3-(chloromethyl)furano[e]indoline (1) and 7-benzyloxy-N-Boc-3-(chloromethyl)furano[f]quinoline (2). DFT calculations indicated that the indoline radical (6B), formed from 5-exo-trig closure of intermediate 6A, was less polar than the corresponding quinoline radical (6C). Based on this result, we report herein that cyclization of chlorides 6–8 in a polar aromatic solvent consistently and exclusively produced the indoline product in good yield (74–83%). [Copyright &y& Elsevier]
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- 2013
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31. A non-reactive natural product precursor of the duocarmycin family has potent and selective antimalarial activity.
- Author
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Alder A, Struck NS, Xu M, Johnson JW, Wang W, Pallant D, Cook MA, Rambow J, Lemcke S, Gilberger TW, and Wright GD
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- Animals, DNA chemistry, Duocarmycins, Humans, Antimalarials pharmacology, Biological Products pharmacology, Folic Acid Antagonists, Malaria, Parasites
- Abstract
We identify a selective nanomolar inhibitor of blood-stage malarial proliferation from a screen of microbial natural product extracts. The responsible compound, PDE-I
2 , is a precursor of the anticancer duocarmycin family that preserves the class's sequence-specific DNA binding but lacks its signature DNA alkylating cyclopropyl warhead. While less active than duocarmycin, PDE-I2 retains comparable antimalarial potency to chloroquine. Importantly, PDE-I2 is >1,000-fold less toxic to human cell lines than duocarmycin, with mitigated impacts on eukaryotic chromosome stability. PDE-I2 treatment induces severe defects in parasite nuclear segregation leading to impaired daughter cell formation during schizogony. Time-of-addition studies implicate parasite DNA metabolism as the target of PDE-I2 , with defects observed in DNA replication and chromosome integrity. We find the effect of duocarmycin and PDE-I2 on parasites is phenotypically indistinguishable, indicating that the DNA binding specificity of duocarmycins is sufficient and the genotoxic cyclopropyl warhead is dispensable for the parasite-specific selectivity of this compound class., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 Elsevier Ltd. All rights reserved.)- Published
- 2022
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32. The effect of a bromide leaving group on the properties of nitro analogs of the duocarmycins as hypoxia-activated prodrugs and phosphate pre-prodrugs for antitumor therapy
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Stevenson, Ralph J., Denny, William A., Ashoorzadeh, Amir, Pruijn, Frederik B., van Leeuwen, Wouter F., and Tercel, Moana
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- *
ANTINEOPLASTIC antibiotics , *PRODRUGS , *BROMIDES , *HYPOXEMIA , *ALKYLATING agents , *NITRO compounds , *PHOSPHATES , *THERAPEUTICS - Abstract
Abstract: Nitro seco analogs (nitroCBIs) of the antitumor antibiotic duocarmycins are a new class of hypoxia activated prodrugs. These compounds undergo hypoxia-selective metabolism to form potent DNA alkylating agents. A series of four nitroCBI alcohol prodrugs containing a bromide rather than chloride or sulfonate leaving group was synthesized. In assays for in vitro hypoxia-selective cytotoxicity against human tumor cell lines the two bromides with DNA minor groove binding basic side chains displayed hypoxic cytotoxicity ratios (HCRs) of 52–286 in HT29 cells and 41–43 in SiHa cells. These values compare well with a related previously reported chloride analog. The corresponding more water soluble phosphate pre-prodrugs of the bromides were synthesized and evaluated for in vivo antitumor activity against SiHa human tumor xenografts. All four phosphates, with both neutral and basic side chains, demonstrated activity providing statistically significant hypoxic log10 cell kills of 0.87–2.80 at non-toxic doses, matching or proving superior to those of their chloride analogs. [Copyright &y& Elsevier]
- Published
- 2011
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33. A novel achiral seco-cyclopropylpyrido[e]indolone (CPyI) analog of CC-1065 and the duocarmycins: Synthesis, DNA interactions, in vivo anticancer and anti-parasitic evaluation
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Chavda, Sameer, Babu, Balaji, Yanow, Stephanie K., Jardim, Armando, Spithill, Terry W., Kiakos, Konstantinos, Kluza, Jerome, Hartley, John A., and Lee, Moses
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- *
ANTINEOPLASTIC agents , *CHIRALITY , *CHEMICAL bonds , *ADENINE , *PROTOZOAN diseases , *THERMAL analysis , *LABORATORY mice , *CC-1065 (Drug) - Abstract
Abstract: The synthesis of an achiral seco-hydroxy-aza-CBI-TMI analog (8) of the duocarmycins is reported. Its specificity for the DNA minor groove of AT-rich sequences and covalent bonding to adenine-N3 was ascertained by a thermal cleavage assay. Compound 8 was found to be cytotoxic in the nanomolar range against murine and human cancer cells. It was further demonstrated that compound 8 was active against murine melanoma (B16-F0) grown in C57BL/6 mice. Compound 8 was also shown to inhibit the growth of the protozoan parasites Leishmania donovani, Leishmania mexicana, Trypanosoma brucei, and Plasmodium falciparum in culture. [Copyright &y& Elsevier]
- Published
- 2010
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34. Synthesis and evaluation of duocarmycin SA analogs incorporating the methyl 1,2,8,8a-tetrahydrocyclopropa[c]oxazolo[2,3-e]indol-4-one-6-carboxylate (COI) alkylation subunit
- Author
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Boyle, Kristopher E., MacMillan, Karen S., Ellis, David A., Lajiness, James P., Robertson, William M., and Boger, Dale L.
- Subjects
- *
ORGANIC synthesis , *PYRROLES , *CARBOXYLIC acids , *DRUG design , *OXAZOLES , *DNA , *ANTINEOPLASTIC agents , *ALKYLATING agents , *THERAPEUTICS - Abstract
Abstract: The design, synthesis and evaluation of methyl 1,2,8,8a-tetrahydrocyclopropa[c]oxazolo[2,3-e]indol-4-one-6-carboxylate (COI) derivatives are detailed representing analogs of duocarmycin SA containing an oxazole replacement for the fused pyrrole found in the alkylation subunit. [Copyright &y& Elsevier]
- Published
- 2010
- Full Text
- View/download PDF
35. Investigation of the transformations of a novel anti-cancer agent combining HPLC, HPLC–MS and direct ESI–HRMS analyses.
- Author
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Tietze, Lutz F., Krewer, Birgit, and Frauendorf, Holm
- Subjects
- *
PRODRUGS , *ANTINEOPLASTIC agents , *CANCER treatment , *HIGH performance liquid chromatography , *ALKYLATION - Abstract
One of the main problems of anti-cancer therapy is an insufficient differentiation between normal and malignant cells by the known anti-proliferant agents. The antibody-directed enzyme prodrug therapy is a promising approach for a selective treatment of cancer, in which a non-toxic prodrug is enzymatically converted into a highly cytotoxic drug at the surface of malignant cells by a targeted antibody–enzyme conjugate. The transformations and the stability of a very promising novel prodrug and its corresponding cytotoxic derivative were now investigated in detail by high-performance liquid chromatography (HPLC)–mass spectrometry (MS). In order to determine the time-dependent DNA alkylation efficiency and the sequence selectivity of the novel compounds, DNA binding studies using direct electrospray–Fourier transform ion cyclotron resonance–MS (ESI–FTICR–MS) have been performed. These measurements were accompanied by HPLC analyses followed by MS of the separated species to confirm the results of the direct ESI–FTICR–MS measurements. The sites of DNA alkylation could be identified unambiguously by the mass spectrometric fragmentation pattern of the alkylated oligodeoxynucleotides as well as by the results of HPLC followed by MS. A combination of all techniques applied led to a better understanding of the mode of action of the new therapeutics and might be used for an estimation of the cytotoxicity of different prodrugs and drugs since the alkylation efficiency correlates with the bioactivity of the compounds in cell culture investigations. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]
- Published
- 2009
- Full Text
- View/download PDF
36. Antibody-Directed Enzyme Prodrug Therapy: A Promising Approach for a Selective Treatment of Cancer Based on Prodrugs and Monoclonal Antibodies.
- Author
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Tietze, Lutz F. and Krewer, Birgit
- Subjects
- *
ANTIBODY-directed enzyme prodrug therapy , *CANCER treatment , *MONOCLONAL antibodies , *IMMUNOGLOBULINS , *DRUG therapy - Abstract
The antibody-directed enzyme prodrug therapy allows a selective liberation of cytotoxic agents from non-toxic prodrugs in cancerous tissue by targeted antibody–enzyme conjugates. We have developed a series of novel glycosidic prodrugs based on the natural antibiotic CC-1065 and the duocarmycins, which are up to 4800 times less toxic than the drugs liberated from these prodrugs in the presence of the activating enzyme (e.g., β-d-galactosidase). Furthermore, the drugs show very high cytotoxicities with IC50 values of as low as 4.5 pm. In this report, we summarize our recent results on the development and biological evaluation of these novel third-generation prodrugs with higher water solubility, higher difference in cytotoxicity between the prodrugs and the corresponding drugs and improved cytotoxicity of the drugs as compared with previous compounds. [ABSTRACT FROM AUTHOR]
- Published
- 2009
- Full Text
- View/download PDF
37. Design, synthesis, and biological testing of pyrazoline derivatives of combretastatin-A4
- Author
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Johnson, Marlie, Younglove, Brent, Lee, Lauren, LeBlanc, Regan, Holt, Herman, Hills, Patrice, Mackay, Hilary, Brown, Toni, Mooberry, Susan L., and Lee, Moses
- Subjects
- *
ACETYLATION , *CELL lines , *MICROTUBULES , *POLYMERIZATION - Abstract
Abstract: Fourteen N-acetylated and non-acetylated 3,4,5-tri- or 2,5-dimethoxypyrazoline analogs of combretastatin-A4 (1) were synthesized. A non-acetylated derivative (5a) with the same substituents as CA-4 (1) was the most active compound in the series, with IC50 values of 2.1 and 0.5μM in B16 and L1210 cell lines, respectively. In contrast, a similar compound with an acetyl group at N1 of the pyrazoline ring (6g) showed poor activity in the cell lines studied. A cell-based assay indicated that compound 5a caused extensive microtubule depolymerization with an EC50 value of 7.1μM in A-10 cells while no activity was seen with the acetylated compound. Molecular modeling studies showed that these compounds possess a twisted conformation similar to CA-4 (1). [Copyright &y& Elsevier]
- Published
- 2007
- Full Text
- View/download PDF
38. Binding of f-PIP, a pyrrole- and imidazole-containing triamide, to the inverted CCAAT box-2 of the topoisomerase IIα promoter and modulation of gene expression in cells
- Author
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Le, N. Minh, Sielaff, Alan M., Cooper, Amanda J., Mackay, Hilary, Brown, Toni, Kotecha, Minal, O’Hare, Caroline, Hochhauser, Daniel, Lee, Moses, and Hartley, John A.
- Subjects
- *
DNA topoisomerase II , *ISOMERASES , *GENES , *MOLECULAR genetics - Abstract
Abstract: An N-formamido pyrrole- and imidazole-containing triamide (f-PIP) has been shown by DNase I footprinting, SPR, and CD studies to bind as a stacked dimer to its cognate sequences: 5′-TACGAT-3′ (5′-flank of the inverted CCAAT box-2 of the human topoisomerase IIα promoter) and 5′-ATCGAT-3′. A gel shift experiment provided evidence for f-PIP to inhibit protein–DNA interaction at the ICB2 site. Western blot studies showed that expression of the topoisomerase IIα gene in confluent NIH 3T3 cells was induced by treatment with f-PIP. The results suggested that the triamide was able to enter the nucleus, interacted with the target site within ICB2, inhibited NF-Y binding, and activated gene expression. [Copyright &y& Elsevier]
- Published
- 2006
- Full Text
- View/download PDF
39. DNA interstrand crosslinking agents: Synthesis, DNA interactions, and cytotoxicity of dimeric achiral seco-amino-CBI and conjugates of achiral seco-amino-CBI with pyrrolobenzodiazepine (PBD)
- Author
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Purnell, Bethany, Sato, Atsushi, O’Kelley, Amanda, Price, Carly, Summerville, Kaitlin, Hudson, Stephen, O’Hare, Caroline, Kiakos, Konstantinos, Asao, Tetsuji, Lee, Moses, and Hartley, John A.
- Subjects
- *
DNA , *GENES , *NUCLEIC acids , *APOPTOSIS - Abstract
Abstract: The design and synthesis of three novel bisalkylating agents derived from the achiral seco-duocarmycin or CC-1065 analogs and pyrrolobenzodiazepines (PBDs) are described: achiral seco-CBI (cyclopropanebenz[e]indoline)-PBD 11, achiral seco-CI-PBD 12, and achiral seco-CBI dimer 13. Compounds 11 and 12 demonstrated enhanced cytotoxicity over the monomer counterparts against the growth of P815 murine mastocytoma cells in culture. Conjugate 11 was found to covalently react with adenine-N3 positions within the minor groove at AT-rich sequences and to produce DNA interstrand crosslinks. Both compounds were found to induce apoptosis in P815 cells. Due to its poor water solubility, dimer 13 did not give any appreciable DNA binding or cytotoxicity. [Copyright &y& Elsevier]
- Published
- 2006
- Full Text
- View/download PDF
40. SYD985, a novel duocarmycin-based HER2-targeting antibody-drug conjugate, shows promising antitumor activity in epithelial ovarian carcinoma with HER2/Neu expression
- Author
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Pei Hui, Dan-Arin Silasi, Alessandro D. Santin, Babak Litkouhi, Luca Zammataro, Elena Ratner, Elena Bonazzoli, Gary Altwerger, Alice Masserdotti, Masoud Azodi, Peter E. Schwartz, Natalia Buza, Gloria S. Huang, Stefania Bellone, Francesca Pettinella, Serena Wong, Gulden Menderes, and Jonathan Black
- Subjects
0301 basic medicine ,Indoles ,endocrine system diseases ,Receptor, ErbB-2 ,Mice, SCID ,Carcinoma, Ovarian Epithelial ,Ado-Trastuzumab Emtansine ,HER2/neu ,Mice ,Random Allocation ,chemistry.chemical_compound ,0302 clinical medicine ,Cytotoxic T cell ,Medicine ,Neoplasms, Glandular and Epithelial ,Cytotoxicity ,Ovarian Neoplasms ,Antibody-dependent cell-mediated cytotoxicity ,biology ,Immunotoxins ,Obstetrics and Gynecology ,Middle Aged ,Pyrrolidinones ,female genital diseases and pregnancy complications ,Oncology ,030220 oncology & carcinogenesis ,Female ,Adult ,musculoskeletal diseases ,congenital, hereditary, and neonatal diseases and abnormalities ,Antibody-drug conjugate ,Antibodies, Monoclonal, Humanized ,Article ,Duocarmycins ,03 medical and health sciences ,In vivo ,Cell Line, Tumor ,Animals ,Humans ,Maytansine ,Antineoplastic Agents, Alkylating ,Duocarmycin ,Aged ,business.industry ,Antibody-Dependent Cell Cytotoxicity ,Bystander Effect ,Trastuzumab ,medicine.disease ,Xenograft Model Antitumor Assays ,030104 developmental biology ,chemistry ,Immunology ,biology.protein ,Cancer research ,business ,Ovarian cancer - Abstract
Epithelial ovarian cancer (EOC) is an aggressive and heterogeneous disease.10% of EOC demonstrate HER2/neu 3+ receptor over-expression. However, moderate to low (i.e., 2+ and 1+) HER2/neu expression is reported in up to 50% of EOC. The objective of this study was to compare the anti-tumor activity of SYD985, a novel HER2-targeting antibody-drug conjugate (ADC), to trastuzumab emtansine (T-DM1) in EOC models with differential HER2/neu expression.The cytotoxicity of SYD985 and T-DM1 was evaluated using ten primary EOC cell lines with 0/1+, 2+, and 3+ HER2/neu expression in antibody-dependent cellular cytotoxicity (ADCC), proliferation, viability and bystander killing experiments. Finally, the in vivo activity of SYD985 and T-DM1 was also studied in ovarian cancer xenografts.SYD985 and T-DM1 induced similar ADCC in the presence of peripheral blood lymphocytes (PBL) against EOC cell lines with differential HER2/neu expression. In contrast, SYD985 was 3 to 42 fold more cytotoxic in the absence of PBL when compared to T-DM1 (p0.0001). Unlike T-DM1, SYD985 induced efficient bystander killing of HER2/neu 0/1+ tumor cells when admixed with HER2/neu 3+ EOC cells. In vivo studies confirmed that SYD985 is significantly more active than T-DM1 against HER2/neu 3+ EOC xenografts.SYD985 is a novel ADC with remarkable activity against EOC with strong (3+) as well as moderate to low (i.e., 2+ and 1+) HER2/neu expression. SYD985 is more potent than T-DM1 in comparative experiments and unlike T-DM1, it is active against EOC demonstrating moderate/low or heterogeneous HER2/neu expression.
- Published
- 2017
41. Toxicity and repair of DNA adducts produced by the natural product yatakemycin
- Author
-
Rongxin Shi, Brandt F. Eichman, and Elwood A. Mullins
- Subjects
0301 basic medicine ,Indoles ,DNA Repair ,DNA repair ,DNA damage ,Biology ,010402 general chemistry ,01 natural sciences ,Genome ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,DNA Adducts ,Duocarmycins ,Drug Resistance, Bacterial ,Pyrroles ,Molecular Biology ,Duocarmycin ,Biological Products ,Natural product ,Molecular Structure ,Cell Biology ,0104 chemical sciences ,030104 developmental biology ,chemistry ,Biochemistry ,DNA glycosylase ,Toxicity ,DNA ,DNA Damage - Abstract
Yatakemycin (YTM) is an extraordinarily toxic DNA alkylating agent with potent antimicrobial and antitumor properties and is the most recent addition to the CC-1065 and duocarmycin family of natural products. Though bulky DNA lesions the size of those produced by YTM are normally removed from the genome by the nucleotide-excision repair (NER) pathway, YTM adducts are also a substrate for the bacterial DNA glycosylases AlkD and YtkR2, unexpectedly implicating base-excision repair (BER) in their elimination. The reason for the extreme toxicity of these lesions and the molecular basis for the way they are eliminated by BER have been unclear. Here, we describe the structural and biochemical properties of YTM adducts that are responsible for their toxicity, and define the mechanism by which they are excised by AlkD. These findings delineate an alternative strategy for repair of bulky DNA damage and establish the cellular utility of this pathway relative to that of NER.
- Published
- 2017
42. Influence of a Basic Side Chain on the Properties of Hypoxia-Selective Nitro Analogues of the Duocarmycins: Demonstration of Substantial Anticancer Activity in Combination with Irradiation or Chemotherapy
- Author
-
Ho H. Lee, Sunali Mehta, H. D. Sarath Liyanage, Sally Y. Bai, Frederik B. Pruijn, Moana Tercel, and Jean-Jacques Youte Tendoung
- Subjects
Indoles ,Stereochemistry ,medicine.medical_treatment ,Mice, Nude ,Antineoplastic Agents ,Pharmacology ,01 natural sciences ,Duocarmycins ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,In vivo ,Cell Line, Tumor ,Morpholine ,Drug Discovery ,medicine ,Side chain ,Animals ,Humans ,Ovarian Neoplasms ,Chemotherapy ,010405 organic chemistry ,Chemistry ,Ovary ,Nitro Compounds ,Combined Modality Therapy ,Cell Hypoxia ,Pyrrolidinones ,In vitro ,Gemcitabine ,0104 chemical sciences ,Docetaxel ,030220 oncology & carcinogenesis ,Nitro ,Molecular Medicine ,Female ,Drug Screening Assays, Antitumor ,medicine.drug - Abstract
A new series of nitro analogues of the duocarmycins was prepared and evaluated for hypoxia-selective anticancer activity. The compounds incorporate 13 different amine-containing side chains designed to bind in the minor groove of DNA while spanning a wide range of base strength from pKa 9.64 to 5.24. The most favorable in vitro properties were associated with strongly basic side chains, but the greatest in vivo antitumor activity was found for compounds containing a weakly basic morpholine. This applies to single-agent activity and for activity in combination with irradiation or chemotherapy (gemcitabine or docetaxel). In combination with a single dose of γ irradiation 50 at 42 μmol/kg eliminated detectable clonogens in some SiHa cervical carcinoma xenografts, and in combination with gemcitabine using a well-tolerated multidose schedule, the same compound caused regression of all treated A2780 ovarian tumor xenografts. In the latter experiment, three of seven animals receiving the combination treatment we...
- Published
- 2017
43. Unified Biosynthetic Origin of the Benzodipyrrole Subunits in CC-1065
- Author
-
Hua Yuan, Gong-Li Tang, Sheng Wu, Xiao-Hong Jian, Juan Zhao, Ling-Yun Wang, Wen-Bing Jin, and Yue Yin
- Subjects
0301 basic medicine ,Indoles ,Stereochemistry ,Protein subunit ,Mutant ,Biology ,01 natural sciences ,Biochemistry ,Cyclopropane ,Serine ,Duocarmycins ,03 medical and health sciences ,chemistry.chemical_compound ,Gene cluster ,Pyrroles ,Tyrosine ,Cytotoxicity ,Gene ,Antibiotics, Antineoplastic ,010405 organic chemistry ,General Medicine ,Biosynthetic Pathways ,0104 chemical sciences ,030104 developmental biology ,chemistry ,Cyclization ,Molecular Medicine - Abstract
CC-1065 is the first characterized member of a family of naturally occurring antibiotics including yatakemycin and duocarmycins with exceptionally potent antitumor activity. CC-1065 contains three benzodipyrroles (1a-, 1b-, and 1c-) of which the 1a-subunit is remarkable by being composed of a cyclopropane ring, and the mechanism for the biological formation of benzodipyrrole rings remains elusive. Previously, biosynthetic studies of CC-1065 were limited to radioactively labeled precursor feeding experiments, which showed that tyrosine (Tyr) and serine (Ser) were incorporated into the two benzodipyrrole (1b- and 1c-) subunits via the same mode but that this was different from the key cyclopropabenzodipyrrole (1a-) subunit with N1-C2-C3 derived from Ser. Herein, the biosynthetic gene cluster of CC-1065 has been cloned, analyzed, and characterized by a series of gene inactivations. Significantly, a key intermediate bearing a C7-OH group derived from a Δc10C mutant exhibited improved cytotoxicity. Moreover, this data inspired us to suspect that the 1a-subunit might employ the same precursor incorporation mode as the 1b- and 1c-subunits. Subsequently
- Published
- 2017
44. TCR-like antibody drug conjugates mediate killing of tumor cells with low peptide/HLA targets
- Author
-
Christian E Herrera, Camille K Bivens, Amanda L McCormick, Devin B. Lowe, Alexis S Mobley, Jon Weidanz, Manoj K. Sabnani, Timea Wichner, and Laurence M. Wood
- Subjects
0301 basic medicine ,Antibody-drug conjugate ,Indoles ,Antibodies, Neoplasm ,medicine.medical_treatment ,Immunology ,Receptors, Antigen, T-Cell ,Human leukocyte antigen ,Biology ,Duocarmycins ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,Drug Delivery Systems ,0302 clinical medicine ,Antigen ,HLA Antigens ,Cell Line, Tumor ,Neoplasms ,medicine ,Animals ,Humans ,Immunology and Allergy ,Duocarmycin ,T-cell receptor ,Immunotherapy ,Molecular biology ,Pyrrolidinones ,body regions ,030104 developmental biology ,chemistry ,030220 oncology & carcinogenesis ,Cancer cell ,Cancer research ,biology.protein ,Brief Reports ,Antibody ,Peptides - Abstract
The currently marketed antibody-drug conjugates (ADC) destabilize microtubule assembly in cancer cells and initiate apoptosis in patients. However, few tumor antigens (TA) are expressed at high densities on cancer lesions, potentially minimizing the therapeutic index of current ADC regimens. The peptide/human leukocyte antigen (HLA) complex can be specifically targeted by therapeutic antibodies (designated T cell receptor [TCR]-like antibodies) and adequately distinguish malignant cells, but has not been the focus of ADC development. We analyzed the killing potential of TCR-like ADCs when cross-linked to the DNA alkylating compound duocarmycin. Our data comprise proof-of-principle results that TCR-like ADCs mediate potent tumor cytotoxicity, particularly under common scenarios of low TA/HLA density, and support their continued development alongside agents that disrupt DNA replication. Additionally, TCR-like antibody ligand binding appears to play an important role in ADC functionality and should be addressed during therapy development to avoid binding patterns that negate ADC killing efficacy.
- Published
- 2017
45. Sequence specific recognition of DNA by tailor-made hairpin conjugates of achiral seco-cyclopropaneindoline-2-benzofurancarboxamide and pyrrole–imidazole polyamides
- Author
-
Price, Carly A., Lingerfelt, Brian M., Handl, Heather L., Kiakos, Konstantinos, Hartley, John A., and Lee, Moses
- Subjects
- *
DNA , *PYRROLES , *ADENINE , *AMINO acids , *NUCLEOTIDE sequence - Abstract
Abstract: Hairpin conjugates of achiral seco-cyclopropaneindoline-2-benzofurancarboxamide (achiral seco-CI-Bf) and three diamides (ImPy 1, PyIm 2, and PyPy 3, where Py is pyrrole, and Im is imidazole), linked by a γ-aminobutyrate group, were synthesized. The sequence-specific covalent alkylation of the achiral CI moiety with adenine-N3 in the minor groove was ascertained by thermally induced DNA cleavage experiments. The results provide evidence that hairpin conjugates of achiral seco-CI-Bf-γ-polyamides could be tailored to target specific DNA sequences according to a set of general rules: the achiral CI moiety selectively reacts with adenine-N3, a stacked pair of imidazole/benzofuran prefers a G/C base pair, and a pyrrole/benzofuran prefers an A/T or T/A base pair. Models for the binding of hairpin conjugates 1–3 with sequences 5′-TCA(888)G-3′, 5′-CAA(857)C-3′, and 5′-TTA(843)C-3′ are proposed. [Copyright &y& Elsevier]
- Published
- 2005
- Full Text
- View/download PDF
46. A novel class of achiral seco-analogs of CC-1065 and the duocarmycins: design, synthesis, DNA binding, and anticancer properties
- Author
-
Kupchinsky, Stanley, Centioni, Sara, Howard, Tiffany, Trzupek, John, Roller, Shane, Carnahan, Virginia, Townes, Heather, Purnell, Bethany, Price, Carly, Handl, Heather, Summerville, Kaitlin, Johnson, Kimberly, Toth, James, Hudson, Stephen, Kiakos, Konstantinos, Hartley, John A., and Lee, Moses
- Published
- 2004
- Full Text
- View/download PDF
47. Synthesis and evaluation of N-aryl and N-alkenyl CBI derivatives
- Author
-
Parrish, Jay P., Trzupek, John D., Hughes, Terry V., Hwang, Inkyu, and Boger, Dale L.
- Published
- 2004
- Full Text
- View/download PDF
48. A Novel Tumor-Activated Prodrug Strategy Targeting Ferrous Iron Is Effective in Multiple Preclinical Cancer Models
- Author
-
Lidia Sambucetti, Aras N. Mattis, James A. Wells, Adam R. Renslo, Yihui Shi, Byron Hann, Shaun D. Fontaine, and Benjamin Spangler
- Subjects
0301 basic medicine ,Indoles ,Cell Survival ,Mice, Nude ,Antineoplastic Agents ,Mice, SCID ,Pharmacology ,Article ,Duocarmycins ,Mice ,Structure-Activity Relationship ,03 medical and health sciences ,chemistry.chemical_compound ,Drug Delivery Systems ,Therapeutic index ,Mice, Inbred NOD ,Drug Discovery ,Tumor Cells, Cultured ,medicine ,Animals ,Humans ,Prodrugs ,Pyrroles ,Ferrous Compounds ,Duocarmycin ,Cell Proliferation ,Tumor microenvironment ,Dose-Response Relationship, Drug ,Molecular Structure ,Chemistry ,Mammary Neoplasms, Experimental ,Cancer ,Prodrug ,medicine.disease ,3. Good health ,030104 developmental biology ,Cancer cell ,Drug delivery ,Molecular Medicine ,Female ,Drug Screening Assays, Antitumor ,Conjugate - Abstract
Here we describe a new approach for tumor targeting in which augmented concentrations of Fe(II) in cancer cells and/or the tumor microenvironment triggers drug release from an Fe(II)-reactive prodrug conjugate. The 1,2,4-trioxolane scaffold developed to enable this approach can in principle be applied to a broad range of cancer therapeutics and is illustrated here with Fe(II)-targeted forms of a microtubule toxin and a duocarmycin-class DNA-alkylating agent. We show that the intrinsic reactivity/toxicity of the duocarmycin analog is masked in the conjugated form and this greatly reduced toxicity in mice. This in turn permitted elevated dosing levels, leading to higher systemic exposure and a significantly improved response in tumor xenograft models. Overall our results suggest that Fe(II)-dependent drug delivery via trioxolane conjugates could have significant utility in expanding the therapeutic index of a range of clinical and preclinical stage cancer chemotherapeutics.
- Published
- 2016
49. Analogues of DNA minor groove cross-linking agents incorporating aminoCBI, an amino derivative of the duocarmycins: Synthesis, cytotoxicity, and potential as payloads for antibody–drug conjugates
- Author
-
Ho H. Lee, Jun Guo, Anna C. Giddens, Thomas H. Pillow, Christian K. Miller, Moana Tercel, Guo-Liang Lu, and Gail Lewis Phillips
- Subjects
0301 basic medicine ,Antibody-drug conjugate ,Indoles ,Stereochemistry ,Clinical Biochemistry ,Pharmaceutical Science ,Pyrrolobenzodiazepine ,Antineoplastic Agents ,01 natural sciences ,Biochemistry ,Antibodies ,Duocarmycins ,Structure-Activity Relationship ,03 medical and health sciences ,chemistry.chemical_compound ,Cell Line, Tumor ,Drug Discovery ,Humans ,Structure–activity relationship ,Bifunctional ,Cytotoxicity ,Molecular Biology ,Amination ,Duocarmycin ,Cell Proliferation ,Dose-Response Relationship, Drug ,Molecular Structure ,010405 organic chemistry ,Organic Chemistry ,Cell Cycle Checkpoints ,Pyrrolidinones ,0104 chemical sciences ,Cross-Linking Reagents ,030104 developmental biology ,chemistry ,Molecular Medicine ,Drug Screening Assays, Antitumor ,DNA - Abstract
A Pd-catalysed amination method is used to convert seco-CBI, a synthetic analogue of the alkylating subunit of the duocarmycin natural products, from the phenol to amino form. This allows efficient enantioselective access to the more potent S enantiomer of aminoCBI and its incorporation into analogues of DNA minor groove cross-linking agents. Evaluation in a panel of nine human tumour cell lines shows that the bifunctional agents containing aminoCBI are generally less cytotoxic than their phenolCBI analogues and more susceptible to P-glycoprotein-mediated resistance. However, all bifunctional agents are potent cytotoxins, some in the sub-pM IC50 range, with in vitro properties that compare favourably with established microtubule-targeted ADC payloads.
- Published
- 2016
50. A Phase II Pilot Study of KW-2189 in Patients with Advanced Renal Cell Carcinoma.
- Author
-
Small, Eric, Figlin, Robert, Petrylak, Daniel, Vaughn, David, Sartor, Oliver, Horak, Ivan, Pincus, Rosemarie, Kremer, Alton, and Bowden, Chris
- Abstract
Background: KW-2189 is a semi-synthetic, water-soluble analog of duocarmycin B
2 , a new class of potent antitumor antibiotics produced by streptomyces, with improved in vitro antitumor potency. Patients and methods: Forty patients with pathologically confirmed metastatic renal cell carcinoma were treated in this multicenter, open-label phase II trial. All patients received 0.4mg/m2 KW-2189 as an IV infusion for Cycle 1. Cycles were repeated every 5 to 6 weeks with escalations to 0.5 mg/m2 in the absence of significant toxicity or disease progression. Results:No patient had an objective response. The most common drug-related toxicity was hematological-delayed neutropenia and thrombocytopenia, with recovery by week 6. Non-hematologic toxicity consisted of mild to moderate fatigue, nausea and vomiting, and anorexia that was generally manageable. Conclusions: KW-2189 in this dose and schedule has a predictable safety profile of reversible myelosuppression. No activity in metastatic renal cell carcinoma was demonstrated. [ABSTRACT FROM AUTHOR]- Published
- 2000
- Full Text
- View/download PDF
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