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OUR SCIENCE

CASE 1

The establishment of high resolution imaging mass spectrometry (IMS) for absolute quantification of molecules and compounds in tissue sections

  • Yukari Tanaka
    Yukari Tanaka
    Team leader, Drug Metabolism & Pharmacokinetics, Research Laboratory for Development
  • case_01_photo_02
    Masayuki Sugimoto
    Obesity & Metabolic Diseases, Drug Discovery & Disease Research Laboratory

MS is a useful approach to visualize the localization of endogenous molecules and administered compounds by using a mass spectrometer, without using any probes. It is hoped that this technology will be useful for elucidating the MoA for drug efficacy and toxicity, as well as for the analysis of drug distribution in tissues, and finding biomarkers by localized analysis of endogenous molecules, such as lipids, that are difficult to visualize with existing methods because of difficulties in the specific and quantitative analysis of target molecules.

A Shionogi group including Yukari Tanaka developed the quantitative MALDI-IMS method to resolve these issues, and was successful in analyzing drug distribution in tissues as well as the concentration in regions of interest within tissues. Using this method, Masayuki Sugimoto et al, in collaboration with scientists at Hokkaido University, were successful in visualizing the distributions of sphingomyelin molecular species in the brain sections of mice. Moreover they generated results that led to the identification of new diagnostic markers and analysis of lipid molecule physiological functions by studying the relationship with metabolic enzymes.

The establishment of high resolution imaging mass spectrometry (IMS) for absolute quantification of molecules and compounds in tissue sections

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bibliography

1. Nozomi Takai, Yukari Tanaka, and Hideo Saji

Quantification of small molecule drugs in biological tissue sections by imaging mass spectrometry using surrogate tissue-based calibration standards.
Mass Spectrom (Tokyo). 2014;3(1):A0025.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3968287/

2. Nozomi Takai, Yukari Tanaka, Kazuhiro Inazawa, and Hideo Saji

Quantitative analysis of pharmaceutical drug distribution in multiple organs by imaging mass spectrometry.
Rapid Commun Mass Spectrom. 2012 Jul 15;26(13):1549-56.

http://www.ncbi.nlm.nih.gov/pubmed/22638972

3. Masayuki Sugimoto, Yoichi Shimizu, Takeshi Yoshioka, Masato Wakabayashi, Yukari Tanaka, Kenichi Higashino, Yoshito Numata, Shota Sakai, Akio Kihara, Yasuyuki Igarashi, Yuji Kuge.

Histological analyses by matrix-assisted laser desorption/ionization-imaging mass spectrometry reveal differential localization of sphingomyelin molecular species regulated by particular ceramide synthase in mouse brains.
Biochim Biophys Acta. 2015 Dec;1851(12):1554-65.

http://www.sciencedirect.com/science/article/pii/S1388198115001699

4. Masayuki Sugimoto, Masato Wakabayashi, Yoichi Shimizu, Takeshi Yoshioka, Kenichi Higashino, Yoshito Numata, Tomohiko Okuda, Songji Zhao, Shota Sakai, Yasuyuki Igarashi, Yuji Kuge.

Imaging Mass Spectrometry Reveals Acyl-Chain- and Region-Specific Sphingolipid Metabolism in the Kidneys of Sphingomyelin Synthase 2-Deficient Mice.
PLoS One. 2016 Mar 24;11(3):e0152191.

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0152191

CASE 2

Trojan Horse: The discovery of a new antibiotic, Cefiderocol (S-649266)

  • Yoshinori Yamano
    Yoshinori Yamano
    Chief Scientific Officer for Infectious Diseases, Drug Discovery & Disease Research Laboratory

The lack of antibiotics effective against drug-resistant bacteria is becoming one of the most serious problems in modern medicine, as treatment options are limited. Multidrug-resistant gram-negative bacteria (Pseudomonas aeruginosa, Acinetobacter, and Enterobacteriacea) have been classified by the Center for Disease Control (CDC) in the United States as significant pathogenic bacteria that require an urgent response. The Infectious Disease Research Group led by Yoshinori Yamano at Shionogi is making vigorous efforts to tackle this threat, and has discovered the new antibiotic cefiderocol (S-649266) which has been shown to demonstrate potent efficacy against infections caused by multidrug-resistant Gram-negative bacteria. S-649266 has features which can overcome such resistance that are lacked by existing beta-lactam antibiotics, in particular a siderophore-like structure which enables the formation of iron chelates, in turn facilitating efficient uptake into bacterial cells via active iron uptake pathways. In addition, it has the characteristic of being highly stable for carbapenemases that can degrade a wide range of beta-lactam antibiotics, including carbapenem drugs that are a significant contributing factor to bacteria acquiring multi-drug resistance (MDR). Due to these unique characteristics, we anticipate cefiderocol will be a new treatment option for gram-negative bacterial infections which cannot be treated effectively with existing antibiotics.

Aside from cefiderocol, there are many successful drug discovery programs in the Infectious Diseases area. Examples include the clinical development of S-033188, an anti-influenza drug which acts with a first-in-class mechanism, and the discovery of dolutegravir, which is widely used as an anti-HIV agent. Innovative drug discovery efforts in the Infectious Diseases area are expected to continue steadily in the next decade.

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bibliography

1. Kohira N, West J, Ito A, Ito-Horiyama T, Nakamura R, Sato T, Rittenhouse S, Tsuji M, Yamano Y

In Vitro Antimicrobial Activity of a Siderophore Cephalosporin, S-649266, against Enterobacteriaceae Clinical Isolates, Including Carbapenem-Resistant Strains.
Antimicrob Agents Chemother. 2015 Nov 16;60(2):729-34

http://aac.asm.org/content/60/2/729.long

2. Ito A, Kohira N, Bouchillon SK, West J, Rittenhouse S, Sader HS, Rhomberg PR, Jones RN, Yoshizawa H, Nakamura R, Tsuji M, Yamano Y.

In vitro antimicrobial activity of S-649266, a catechol-substituted siderophore cephalosporin, when tested against non-fermenting Gram-negative bacteria.
J Antimicrob Chemother. 2016 Mar;71(3):670-7

http://jac.oxfordjournals.org/content/71/3/670.long

3. Ito-Horiyama T, Ishii Y, Ito A, Sato T, Nakamura R, Fukuhara N, Tsuji M, Yamano Y, Yamaguchi K, Tateda K.

Stability of Novel Siderophore Cephalosporin S-649266 against Clinically Relevant Carbapenemases.
Antimicrob Agents Chemother. 2016 Jun 20;60(7):4384-6.

http://aac.asm.org/content/60/7/4384.long

4. Itoh, A. et al.

S-649266, a novel siderophore cephalosporin: II. Impact of active transport via iron regulated outer membrane proteins on resistance selection.

F-1563, 54th Intersci Congress Antimicrob Agents Chemother, 2014

CASE 3

Towards suppression of HIV: The development of dolutegravir

  • Takashi Kawasuji
    Takashi Kawasuji
    Department Head of Chemistry Infectious Diseases, Medicinal Chemistry Research Laboratory

The life expectancy of HIV patients has improved with the development of multi-drug combination therapy, but large numbers of tablets, multiple daily doses and the high frequency of side effects can prevent patients from continuing with their medication. Consequently the emergence of drug resistant strains has become a major issue.
There have been efforts at Shionogi for over 20 years to develop anti-HIV agents that can continue to be taken safely while maintaining anti-HIV activity, and which have seen the successful development of the groundbreaking HIV integrase inhibitor, dolutegravir. Takashi Kawasuji's drug discovery team has led the optimization of compounds aimed at both drug efficacy and safety. The main strength of dolutegravir is that the rate of drug resistant strain emergence is markedly lower compared to other HIV integrase inhibitors.

Furthermore, it is small-sized for an anti-HIV drug, and is very easy for patients to take. Administration once a day is possible without taking booster drugs at the same time to maintain the drug concentration in the blood.

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Kawasuji and his team have continued to pursue an ideal form of HIV infection therapy together with pharmacology and virology researchers at Shionogi, while leading many outstanding medicinal chemists. They won the Breakthrough Award, Division of Medicinal Chemistry, PSJ in 2015 in Japan and the American Chemical Society Heroes of Chemistry Award in 2016 in the US for the discovery of dolutegravir.

bibliography

1. Kawasuji T, Johns BA, Yoshida H, Taishi T, Taoda Y, Murai H, Kiyama R, Fuji M, Yoshinaga T, Seki T, Kobayashi M, Sato A, Fujiwara T.

Carbamoyl pyridone HIV-1 integrase inhibitors. 1. Molecular design and establishment of an advanced two-metal binding pharmacophore.
J Med Chem. 2012 Oct 25;55(20):8735-44.

http://pubs.acs.org/doi/abs/10.1021/jm3010459

2. Kawasuji T, Johns BA, Yoshida H, Weatherhead JG, Akiyama T, Taishi T, Taoda Y, Mikamiyama-Iwata M, Murai H, Kiyama R, Fuji M, Tanimoto N, Yoshinaga T, Seki T, Kobayashi M, Sato A, Garvey EP, Fujiwara T.

Carbamoyl pyridone HIV-1 integrase inhibitors. 2. Bi- and tricyclic derivatives result in superior antiviral and pharmacokinetic profiles.
J Med Chem. 2013 Feb 14;56(3):1124-35.

http://pubs.acs.org/doi/abs/10.1021/jm301550c

3. Johns BA, Kawasuji T, Weatherhead JG, Taishi T, Temelkoff DP, Yoshida H, Akiyama T, Taoda Y, Murai H, Kiyama R, Fuji M, Tanimoto N, Jeffrey J, Foster SA, Yoshinaga T, Seki T, Kobayashi M, Sato A, Johnson MN, Garvey EP, Fujiwara T.

Carbamoyl pyridone HIV-1 integrase inhibitors 3. A diastereomeric approach to chiral nonracemic tricyclic ring systems and the discovery of dolutegravir (S/GSK1349572) and (S/GSK1265744).
J Med Chem. 2013 Jul 25;56(14):5901-16.

http://pubs.acs.org/doi/abs/10.1021/jm400645w

4. Kobayashi M, Yoshinaga T, Seki T, Wakasa-Morimoto C, Brown KW, Ferris R, Foster SA, Hazen RJ, Miki S, Suyama-Kagitani A, Kawauchi-Miki S, Taishi T, Kawasuji T, Johns BA, Underwood MR, Garvey EP, Sato A, Fujiwara T.

In Vitro antiretroviral properties of S/GSK1349572, a next-generation HIV integrase inhibitor.
Antimicrob Agents Chemother. 2011 Feb;55(2):813-21.

http://aac.asm.org/content/55/2/813.long