oligosaccharides in drug discovery 2014 am · © glycomar!limited! !!!!! introduction!!...

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Oligosaccharides!in!Drug!Discovery!

!2014!

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© GlycoMar!Limited!www.glycomar.com!

!!!!Introduction!!Glycobiology! is! the! branch! of! biology! concerned! with! the! study! of! structure,! biosynthesis,! and!function! of! saccharides! (sugar! chains),! which! may! exist! purely! or! conjugated! to! other! biological!molecules! to! form! glycoconjugates.! The! study! of! such! molecules! is! technically! challenging! as! no!sequencing! tool,! such! as! that! used! in! proteomics! or! genomics,! is! available.! However,! following! on!from!genomics! and!proteomics,! there! is! increasing! recognition! of! the! importance! of! carbohydrateIbased!molecules! in!basic! cellular!processes.!This!has! resulted! in!more!extensive!glycomic! studies! in!the!areas!of!glycosylation!of! therapeutic!proteins,! glycosylation!patterns! in! cell! recognition,! cellular!glycoprofiling! studies! in! cancer! and! other! diseases,! correlation! between! activity! and! sulphation!patterns! in! glycosaminoglycans,! and! the! improved! chemical! analysis! and! synthesis! of! carbohydrate!molecules.!!Glycobiology!offers!enormous!untapped!potential!in!the!discovery!of!new!therapeutics!derived!from!saccharides!or!other!molecules!which!target!the!biosynthesis!and!function!of!saccharides.!!!!Saccharides:!multifunctional!biological!molecules!!Saccharides! offer! potential! chemical! diversity! orders! of! magnitude! greater! than! their! protein! and!nucleic!acid!counterparts[1].!For!example,!DNA!can!give!256!4Iunit!structures;!amino!acids!can!give!16000!4Iunit!structures,!whereas!the!nine!common!monosaccharides!have!the!theoretical!ability! to!generate! nearly! 16! million! 4Iunit! structures[2].! The! level! of! chemical! information! encoded! in!saccharides!is!therefore!unrivalled.!!

!Graph!taken!from:!!Turnbull!&!Field!!2007!Nature!Chemical!Biology!3(2),!pp74I77!

!Saccharides! have! multiple! functions! which! makes! them! relevant! to! almost! any! area! of! biological!research.!They!are!ubiquitously!present!on!cell!surfaces,!mediating!the!interaction!of!cells!with!other!cells,!with!the!extracellular!matrix!and!with!effector!molecules.!The!glycosylation!of!protein!molecules!is!also!critical!for!their!function,!bioavailability!and!halfIlife.!Polysaccharides!and!glycoconjugates!also!play! a! major! structural! role! in! all! connective! tissues.! Oligosaccharides,! which! are! short! chains!(nominally! 2I10!monosaccharides)! of! sugars! derived! from! larger! polysaccharides! or! synthesised! in!their!own! right,! are!widely! studied! to! identify! functional! groups!within! complex!glycoconjugates!or!polysaccharides.! The! expression! of! saccharides,!which! in# vivo#occurs! by! the! ruleIdriven! action! of! a!multiple! synthetic! enzymes,! is!much!more! complex! than! that! of! proteins! or! nucleotides!which! are!template! driven! processes.! ! All! but! the! very! smallest! saccharides! have! proven! very! difficult! to!

Development of Oligosaccharides in drug discovery

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synthesise! in# vitro! making! current! drug! discovery! and! development! activity! largely! dependent! on!nature!for!the!generation!of!screening!libraries.!!!Oligosaccharide!therapeutics:!!The!central!role!of!saccharides!in!cellular!interactions!means!that!novel!oligosaccharides!are!of!value!in!R&D!programmes!addressing!a!wide!range!of!therapeutic!areas![3,!4],!including:!!

• Inflammation!• Immunity!• Oncology!• Neurodegenerative!disease!• Infection!

!Regulation!of!the!immune!system!is!a!key!area!of!interest!to!glycobiologists,!due!to!the!central!roles!of! carbohydrateIbased! molecules! such! as! glycoproteins! and! glycosaminoglycans! in! cellIcell!recognition,!cell!signalling!and!infection.!With!increasing!understanding!of!the!inflammation!process!in! a! range! of! disorders! including! vascular! disease! (atherosclerosis,! thrombosis,! diabetes),!neurodegeneration! (Alzheimer’s,! Multiple! sclerosis),! cancer! and! orthopaedic! illnesses! (arthritis,!Ankylosing!spondylitis),! there! is!wide!scope! for! the! further!application!of!novel!carbohydrateIbased!drugs.!!!Oligosaccharides-in-use-as-drugs-or-in-development:-!The!use!of!carbohydrateIbased!drugs!is!in!its!infancy,!although!several!are!well!known!(see!Table!1).!In! addition! to! ‘monosaccharideIinspired’! drugs! such! as! influenza! treatment! Tamiflu! (oseltamivir,!Roche),! two! blockbuster! drugs,! Acarbose! (Bayer)! and! heparin,! stand! out.! Both! saccharides! were!isolated,!and!reached!the!clinic,!before!a!detailed!structureIactivity!relationship!had!been!carried!out.!Heparin! is!the!key!example!of!a!major!carbohydrateIbased!molecule,!which!has!been!developed!for!medical!use! in!antiIcoagulant! therapies.!Low!molecular!weight!heparins! (eg.!Certoparin,!Dalteparin)!and!various!derivatives!(Fondaparinux!–!fully!synthetic)!have!been!developed!to!improve!efficacy!and!halfIlive,!and!some!are!now!being!trialled!for!nonIthrombotic/vascular!applications!(eg.!Certoparin!for!inflammatory! aspects! of! Alzheimer’s! disease).! It! is! only! comparatively! recently! that! the! antiIinflammatory!properties!of!heparin!have!been!the!subject!of!drug!discovery!research!&!development.!!Delivery-and-immunogenicity:--The!drugs!listed!in!Table!1!have!overcome!some!of!the!perceived!limitations!of!sugarIbased!molecules!in!terms!of!synthesis,!delivery!and!immunogenicity.!Although!many!still!require!intravenous!delivery,!several! are! available! orally! (e.g.! Pentosan! polysulphate)! and! further! research! is! targeting! the!improvement!of!oral!availability!by!reducing!compounds!to! their!smallest!active!components,!or!by!combining!with!other!molecules!(e.g.!sulodexide).!Improvements!in!synthesis!have!meant!that!some!compounds!can!be!synthesised!(e.g.!Fondaparinux),!and!small!active!components!can!be!selected!or!modified!to!improve!efficacy.!No!major!problems!have!been!reported!with!immunogenicity!either!in!animal!trials!or!as!approved!drugs,!using!synthetic!or!animal!derived!material,!including!that!obtained!from!marine!invertebrates.!!Approval!of!marine!derived!heparinoids!is!more!common!in!China!than!in!Western!Countries!and!there!are!a!number!of!phase!I!and!phase!II!trials!ongoing!in!the!East.!

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Table&1.&Examples&of&carbohydrate6based&drugs&in&use&or&in&development&

Company) Compound) Activity)/)disorder) Development)Numerous) Heparin)and)derivatives:)in)particular)low)

molecular)weight)forms)AntiAcoagulants) Since)1940’s)

GSK) Arixtra)(fondraparinux))–)synthetic)heparin)pentasaccharide)Generic)now)available)(Dr)Reddy’s))

Factor)Xa)inhibitor,)DVT))&)pulmonary)embolism) 1982)

Astellas) Auranofin)(Ridaura):)organogold)monosaccharide) AntiArheumatic)) 1983)GSK) Zanamivir)(Relenza):)modified)monosaccharide) AntiAinfluenza)(neuraminidase)inhibitor)) 1992)Johnson)&)Johnson) Topiramate)(Topamax):)modified)monosaccharide) AntiAepileptic) 1987)Bayer) Acarbose)(Glucobay))(PseudoAoligosaccharide)) Type)II)diabetes)

AlphaAglucosidase,)alphaAamylase)inhibitor.)1990)

OrthoAMcNeil)Janssen)Pharmaceutical)

Elmiron)(Pentosan)polysulphate)) Cystitis)(interest)in)development)for)CJD)) 1996)

Alfa)Wassermann) Sulodexide)(Vessel™):)mixture)of)LMW)heparin)and)dermatan)sulphate)

Various)cardiovascular)indications) Marketed)since)1980’s)

Merion)Pharmaceuticals) MERA102:)uLMW)heparin) Oral)uLMW)heparin)for)DVT) Phase)IIa)Sanofi)Aventis) Idrabiotaparinux,)biotinylated)synthetic)heparin)

pentasaccharide)Biotinylated)idraparinux)for)the)treatment)DVT)and)pulmonary)embolism)

Phase)III)

Cantex)Pharmaceuticals)(previously)ParinGenix))

PGX100:)OAdesulphated)heparin) OAdesulphated)heparin)for)cardiac)ischemiaAreperfusion)injury,)COPD.)Used)in)AML.)

Phase)I/)Phase)II)

Hunter)Fleming)(now)Newron))

HF0420)–)low)molecular)weight)oligosaccharide) Prevention)of)antiAcancer)induced)neuropathy)(neuroprotective))

Discontinued))

Progen)(Australia)))

PIA88)(Phosphomannopentaose)sulphate)) Heparan)sulphate)mimetic)–)AntiAangiogenic/antiAmetastatic.)Adjuvant)therapy)for)postAresection)hepatocellular)carcinoma)

Phase)III)

PG545:)fully)synthetic)heparan)sulphate)mimetic) Heparan)sulphate)mimetic)–)AntiAangiogenic/antiAmetastatic.)

Late)PreAclinical))

Endotis)Pharma))

EP80061)–)synthetic)oligsaccharide) AntiAmetastatic,)heparanase)inhibitor) Discontinued))EP80031)–)synthetic)oligosaccharide) Hematopoietic)stem)cell)mobilization) Discontinued)EP37151)–)oral)fondaparinux) Anticoagulant) Discontinued)EP217609)–)biotinylated)thrombin)inhibitor)coupled)to)pentasaccharide)

Indirect)factor)Xa)and)direct)thrombin)(Factor)IIa))inhibitor)that)can)be)neutralized)quickly)and)effectively)without)rebound)effect)by)a)specific)antidote:)avidin)

Discontinued)

Biotec)Pharmacon) BetaAglucan) Wounds,)Cancer,)IBS,)Asthma)) Preclinical)


Marinova) Fucoidan) Immunity,)Osteoarthritis,)HSVA1,)AntiAoxidant)) Phase)I)&)II)GlycoMar)/)Verona)Pharma) GLY145)–)glycosaminoglycan) AntiAinflammatory) Preclinical)AlgiPharma) Oligo)G)from)alginate) Cystic)Fibrosis)) Phase)I)Wu)et)al)[5]) Heparinoid)911)A)Sulphated)

polymannuroguluronate)(from)alginate))HIV/AIDS) Phase)IIa)

China)Ocean)University*) Polymannuronic)acid)propyl)sulphate)(PMS)) Cerebral)Ischemia.)Influenza)) Phase)IIa)Heparinoid)916)(from)chitosan)) Atherosclerosis) Phase)III)Propylene)glycol)alginate)sodium)sulphate)(PSS)) AntiAangiocardiopathy)) Available)in)China)Propylene)glycol)mannate)sulphate)(PGMS)" Posterior)capsular)opacification) Available)in)China)HS971)–)Oligo)mannuronate)from)alginate) Alzheimer’s)) Phase)IIb)

Sun)et)al[6]) HS203:)oligosaccharide)complex)of)chromium)(III))and)oligomannuronic)acid)

AntiADiabetes) Preclinical))

)*Patent)relating)to)use)of)PMS)in)influenza)appears)to)be)owned)by)the)University.)


Oligosaccharide-libraries-for-drug-discovery-research:--Native)glycoconjugates)and)polysaccharides)have)limited)value)in)drug)discovery)research)due)to)their)large)size)(MW)10)–)1000)kDa))and)structural)heterogeneity,)although)heparin)is)a)notable)historical)exception)which)has)been)used)in)its)native)form)(manufactured)form)bovine)lung)or)porcine)gastric)mucosa).)Oligosaccharides)generated) from)native)molecules)offer) the)opportunity) to) identify)active)functional) subunits) and) of) revealing) cryptic) activity.) Heparin) provides) an) example) of) a) blockbuster)saccharide) that) has) progressed) from) unfractionated) polysaccharide,) to) low) molecular) weight)polysaccharide,)and)now)synthetic)pentasaccharide,)through)the)application)of)glycomic)tools.))Oligosaccharide)libraries)clearly)have)value)for)drug)discovery)research)in)a)wide)range)of)diseases)[7].)Such) libraries) are) becoming) increasingly) available) from) a) small) number) of) specialist) suppliers) (see)Appendix)1)) largely)based)on)glycosaminoglycans)such)as)heparin)and)chondroitin)sulphate,)marine)polysaccharides) such) as) alginate) and) chitosan,) and) oligosaccharides) derived) from) common) plant)polysaccharides.) All) natural) oligosaccharides) are) subject) to) varying) degrees) of) structural) and) size)heterogeneity,)which) pose) problems) in) detailed) structure) activity) studies.) The)wider) availability) of)oligosaccharide) synthesisers) (e.g.) Glyconeer)) will) have) a) significant) impact) on) the) availability) of)oligosaccharides)for)drug)discovery)research,)by)reducing)reliance)on)natural)products)and)improving)the)quality)(homogeneity)/)purity))of)oligosaccharides.))The) unique) structures) and) sulphation) patterns) of) marine) derived) sugars) are) being) increasingly)recognised) and) investigated) [8].) Several) polysaccharides) and) oligosaccharides) are) in) use) as)nutraceuticals,)with)pharmaceutical)development)of)others)in)preclinical)and)clinical)trials,)and)a)few)drugs) on) the) market) in) China.) Depolymerised) and) low) molecular) weight) forms) dominate) clinical)investigations)with)oral)delivery)possible)and)no)reported)problems)with) immunogenicity.)Marinova)(Australia))are)involved)in)the)development)of)fucoidanTbased)products)from)seaweeds)[9],)using)inThouse)modulation)of)sulphation)and)acetylation.)They)are)investigating)applications)in)cardiovascular,)infection,)oncology)and)inflammation)areas,)as)are)several)other)research)and)commercial) labs.)This)parallels)the)activity)by)IFREMER,)France.)Other)compounds)have)been)brought)forward)by)academic)research)groups,)in)particular)a)Brazilian)team)from)Rio)de)Janeiro,)often)in)association)with)industrial)partners)eg.)SanofiTAventis)(sea)squirt)heparin).)A)significant)number)of)oligosaccharide)drugs)derived)from)seaweeds)have)been)developed)by)China)Ocean)University,)including)marketed)drugs.)))Table-2.-Examples-of-marine<derived-carbohydrate<based--therapeutic-molecules-

Compound) Source) Activity) Status)Chondroitin)sulphates) Mussels) Joint)repair)/)maintenance) Nutraceutical)

Galactofucan)sulphate)(GFS)) Seaweed)Marinova,)Australia)

Attenutates)herpes)virus)infections.)Improved)mobilization)

of)hematopoetic)stem)cells)

Nutraceutical)/)Clinical)

Fucosylated)chondroitan)sulphate)

Sea)cucumber) PTselectin)mediated)antiTinflammatory)effects)/)unique)antiTcoagulant)properties)

Preclinical)

Invertebrate)Heparin) Sea)squirt) Arterial)thrombosis) Preclinical)Oligo)G! Alginate) Cystic)Fibrosis)) Phase)I)

Heparinoid)911)T)Sulphated)polymannuroguluronate))

Alginate)(seaweed)) HIV/AIDS) Phase)IIa)

Polymannuronic)acid)propyl)sulphate)(PMS))

Alginate)(seaweed)) Cerebral)Ischemia) Phase)IIa)

Heparinoid)916) Chitosan)(crustacean)shell))

Atherosclerosis) Phase)III)

Propylene)glycol)alginate)sodium)sulphate)(PSS))

Alginate)(seaweed)) AntiTangiocardiopathy)) Available)in)China)

Propylene)glycol)mannate)sulphate)(PGMS))

Alginate)(seaweed)) Posterior)capsular)opacification) Available)in)China)

HS203) Laminaria'japonica) Diabetes) Preclinical))


(seaweed))Heparinoid)971) Seaweed) Alzheimer’s)Disease) Phase)IIb)

)-Glycan-Arrays:--Due)to)the)high)level)of)interest)and)applications)of)glycobiology)in)human)and)animal)medicine)there)has)been)a)concerted)effort)to)develop)glycan)arrays)and)glycochips)for)use)in)biological)and)medical)applications.)Due)to)the)complexity)of)the)carbohydrates)involved)there)has)been)a)perceivable)delay)in)the)development)of)suitable)arrays,)however)more)recent)development)of)oligosaccharide)libraries)have) made) arrays) promising) tools) for) screening) potential) activity) in) novel) compounds) and) are)becoming)an)important)tool)in)drug)discovery.)))Glycan)arrays)provide)a)high)throughput)screening)method)used)to)study)interaction)of)a)large)numbers)of)saccharide)structures)with)target)molecules,)and)can)be)used)to)identify)carbohydrate)interactions)which)may)be)of)value)in)drug)discovery.))The)arrays)are)often)superior)to)screening)techniques)such)as)enzymeT)linked)immunosorbent)assays)(ELISA))as)thousands)of)interactions)can)be)analysed)at)one)time,)and)only)minute)amounts)of)analyte)and)ligand)are)required)for)reliable)results.)Oligosaccharides)which)have)either)been)isolated)or)synthesised)are)covalently)attached)to)glass,)plastic)or)gold)services)or,)are)placed)on)beads)in)fibreT)optic)wells)[7].)More)recent)screening)methods)also)use)oligosaccharides)which)have)been)paired)with)a)linker)on)the)reducing)terminus)and)then)covalently)attached)to)the)slide)–)fluorescent)labelling)is)used)to)highlight)interactions)as)well)as)mass)spectrometry)analysis,)and)DNA)printing)and)scanning)equipment)can)be)used)to)evaluate)the)array.))One)particularly)valuable)example)of)where)glycan)arrays)have)been)employed)is)during)the)discovery)of)HIV)vaccine)candidates)where)epitope)mapping)of)HIVT)related)antibodies)was)performed.)The)information)obtained)was)vital)in)identifying)carbohydrate)based)vaccinations)[7].))--Oligosaccharide-based-drug-discovery:--GlycoMar) is) a) biopharmaceutical) company) uniquely) focused) on) the) development) of) novel) antiTinflammatory) therapies) based) on) saccharide) molecules) from) a) diverse) selection) of) predominantly)marine) organisms.) This) combination) of) diverse) biological) source) with) a) targeted) but) inherently)diverse)chemistry)has)generated)a)hit)rate)100T1000)times)higher)than)conventional)small)molecule)screening.)))))

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References-)1.) Turnbull,)J.E.)and)R.A.)Field,)Emerging'glycomics'technologies.)Nat)Chem)Biol,)2007.)3(2):)p.)

74.)2.) Maeder,)T.,)Sweet'medicines'8'Sugars'play'critical'roles'in'many'cellular'functions'and'in'

disease.'Study'of'those'activities'lags'behind'research'into'genes'and'proteins'but'is'beginning'to'heat'up.'The'discoveries'promise'to'yield'a'new'generation'of'drug'therapies.)Scientific)American,)2002.)287(1):)p.)40T47.)

3.) Koeller,)K.M.,)Emerging'themes'in'medicinal'glycoscience.)Nature)Biotechnology,)2000.)18(8):)p.)835T841.)

4.) Dube,)D.H.)and)C.R.)Bertozzi,)Glycans'in'cancer'and'inflammation'8'potential'for'therapeutics'and'diagnostics.)Nature'Reviews'Drug'Discovery,)2005.)4:)p.)477T488.)

5.) Wu,)Y.L.,)et)al.,)Sulfated'polymannuroguluronate'inhibits'Tat8induced'SLK'cell'adhesion'via'a'novel'binding'site,'a'KKR'spatial'triad.)Acta)Pharmacol)Sin,)2011.)32(5):)p.)647T54.)

6.) Sun,)S.,)et)al.,)Microanalysis'of'oligosaccharide'HS203'in'beagle'dog'plasma'by'postcolumn'fluorescence'derivatization'method.)Carbohydr)Polym,)2012.)89(2):)p.)661T6.)

7.) Seeberger,)P.H.,)Synthesis'and'medical'applications'of'oligosaccharides.)Nature,)2007.)446:)p.)1046T1051.)

8.) Laurienzo,)P.,)Marine'polysaccharides'in'pharmaceutical'applications:'an'overview.)Mar)Drugs,)2010.)8(9):)p.)2435T65.)

9.) Fitton,)J.H.,)Therapies'from'fucoidan;'multifunctional'marine'polymers.)Mar)Drugs,)2011.)9(10):)p.)1731T60.)

)Appendix-1-<-Oligosaccharide-suppliers-)Aglyco)T)http://www.aglyco.com/)Elicityl)T)http://www.elicitylToligotech.com/)Grampian)Enzymes)–)Aberdeen,)UK)Iduron)T)http://www.iduron.co.uk/)Qingdao)BZTOligo)Co)T)http://marineToligo.com/index.html

GlycoMar)is)a)biopharmaceutical)company,)located)at)the)European)Centre)for)Marine)Biotechnology)in)Scotland,)UK.)The)company)has)created)a)unique)combination)of)marine)biology,)glycobiology,)and)drug)discovery.))))))GlycoMar)Limited)European)Centre)for)Marine)Biotechnology)Scottish)Marine)Institute)Dunbeg,)Oban,)Argyll)Scotland)PA37)1QA))Tel:)+44)(0)1631)559)370)Fax:)+44)(0)1631)559)010))Email:)[email protected])))))

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oligosaccharides in drug discovery 2014 am · © glycomar!limited! !!!!! introduction!!...

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