This document discusses cytokines, their properties, categories, and roles in health and disease. It also covers the history of cytokine research and development of cytokine therapies. Specifically, it summarizes: 1) Cytokines are cell signaling proteins that regulate immune and inflammatory responses. They are produced by immune cells and play roles in cancer, infections, and autoimmune diseases. 2) Cytokine research began in the 1950s with the discovery of endogenous pyrogen, nerve growth factor, and interferon. Major discoveries in the 1970s-1980s included TNF, CSFs, and chemokines. 3) Cytokines are now being developed as therapeutic agents to treat diseases like hepatitis C and cancer by enhancing immunity

1. Cytokines in health and
disease and its implication in
new drug development

2. Synopsis
• Introduction
• History
• Properties
• Categories
• cytokines as therapeutic agents
• Anti cytokines as therapeutic agents
• cytokine therapies in clinical trial
• Future perspectives

3. Introduction
• Cytokine - cyto - “cell” – kinin-
‘hormones’.
• Hormone like protein /Bioactive substance
• Stanley cohen in 1974 - term “cytokine”

4. Introduction
intercellular regulatory proteins
• initiating,
• maintaining,
• regulating
immunologic hematopoetic
inflammatory processes.

5. Cytokines
• WBC
• macrophages
• helper T cells
• endothelium
• epithelium
• connective tissue cell

6. Cytokines
• cancer
• metabolic disorders
• autoimmune disorders
• inflammatory diseases –
rheumatoid arthritis , psoriasis,
asthma, Crohn's disease,
multiple sclerosis

7. History

8. History
• Research - began in 1950
• endogenous pyrogen (1953) – Bennet& Beeson
• Nerve growth factor (1953)-
Montalcini&Hamburger
• Interferon (1957)- Isaacs & lindenmann
• MIF (1966) -Johndavid & Barrybloom

9. History
• Lymphokines ( 1969) – Dudley Dumonde
• TNF (1970 s) –Carswell
•
• CSF (1980 s) – Donald Metcalf
• Chemokines ( late 1980 s)
•
• Anticytokine therapy – (1974 ,1975 ,1989) – Simon
Skurkovich
• Antibody to TNF ( 1985) – Bruce Beutler

10. Properties of cytokines

11. Properties
• Pleiotrophy single cytokine many cells
• Redundancy different cytokine similar
function
• Multifunctional same cytokine regulate
different immune system

12. SSCP, TUMKUR 12

13. o
SSCP, TUMKUR 13

14. Other properties
• Autocrine
• Paracrine
• Endocrine

16. Categories of cytokines

17. Categories
1) Interferons
2) Interleukins
3) Tumour Necrosis Factor
4) Transforming Growth Factor
5) Colony Stimulating Factor
6) Growth Factors

18. Others
• chemokines
• Monokines
• Lymphokines

20. Cytokines innate immunity
• TNF,
• IFNγ
• interleukins IL-1β
• IL-4, IL-6, IL-10, IL-12, IL-18,
• TGFβ

21. Cytokines of acquired immunity
• T & B lymphocytes are involved
• B cells - antibody producing plasma cell
• Tcell - helper(CD4) T cell ,cytotoxic
(CD8)Tcell

22. Cytokines of acquired immunity
• Th1 – IL 2,IFN gamma - CMI
• Th2 – IL4,IL5,IL10,IL13 – humoral
immunity
• Th3 – Ig A

24. Cyokines of haematopoiesis
• Myeloid stem cell proliferate & differentiate
in to
Mature granulocyte cells
Mast cell precursur cells
Monocyte macrophage cell
• Play important role in immune response

25. Stimulators of haematopoiesis
• Governed by specific growth factors
IL3 , IL5, IL6,
IL7 ,IL9 , IL 11
GM –CSF,
G –CSF,
M- CSF
Erythropoietin
thrombopoietin

28. Major effects of interleukin-1 (IL-1) and tumor necrosis factor (TNF) in inflammation

29. • Inflammatory
cytokines IL-1, IL-6
and TNF act on
BRAIN and LIVER to
produce symptoms and
ACUTE-PHASE
PROTEINS

30. Cytokines as therapeutic
agents

31. Cytokine therapy
• Advances in understanding - role of
cytokine – in immune & inflammation
• Help to control
• harmful effect of inflammation
• Tissue damaging immune reaction
• Enhance immunity against cancer &
infection

32. Cytokines as therapeutic agents
• Interferons
• Interferons alpha
interferon alpha -2a , 2b
interferon alpha con-1
peg interferon alpha
interferon alpha –n3

33. Cytokines as therapeutic agents
• Interferon beta
interferon beta 1a ,1b
• Interferon gamma
interferon gamma 1b
• Interleukin - 2

34. Interferons
• Naturally occuring glycoprotein
• Antiviral ,anticellular ,antiproliferative
• Classified – cellular origin , receptors

35. Interferons
• Type 1 IFN - alpha, beta
anti proliferative
antiviral
• Type 2 IFN – gamma
immuno modulatory
weak antiviral

36. Interferons
• Secreted by eukaryotic cells - viral
infections, tumors, and other biological
inducers
• clinical benefits - hepatitis, cancers, multiple
sclerosis, and many other diseases

37. Interferons
• Released by macrophages, lymphocytes ,
tissue cells infected with virus
• Diffuse to the surrounding cells
• Bind to receptor of adjacent cell
• Antiviral protein -block viral reproduction
• Prevent spread of virus

40. Interferons
• Janus activated kinus (JAK)
• Signal transducers and activators of
transcription (STAT)
• transcriptional activation of genes -
antiviral , antiproliferative ,
immunomodulatory effect

42. Alpha Interferon-2a & 2b
• Protein chain - 165 amino acids
• recombinant DNA technology
• Non-glycosylated protein
• Short half life
• a large volume of distribution
• a larger reduction in renal clearance.

43. Indications Interferon alpha 2a & 2b
• chronic hepatitis c
• AIDS related kaposis sarcoma
• Chronic myelogenous leukemia
• Hairy cell leukemia

44. Interferon alpha 2a & 2b
• IFN alpha 2b –
malignant melanoma
condyloma acuminata
follicular lymphoma
chronic hepatitis B

45. Pegylated Interferon-2a
• recombinant interferon alpha-2a covalently
conjugated with bis-monomethoxy
polyethylene glycol (PEG)
• First developed by Davis, Abuchowski and
colleagues in the 197

46. Peg interferon – alpha
• Absorption prolonged
• Increases half life
• Increase drug stability
• enhanced drug solublity
• Lower toxicity

47. Peg interferon – alpha
• Advantage –
long half life –once daily
enhanced compliance
superior antiviral activity

48. Peg interferon – alpha
• Dis advantage - high cost
• Indication - treatment of hepatitis c
infection

49. IFN alpha con-1
• Known as concensus IFN
• Synthetic Recombinant product
• Treatment of chronic hepatitis C infection
in relapsers and non responders

50. IFN alpha –n3
• Natural human IFN alpha from human
leukocytes
• Treatment of
refractory condyloma acuminata
• intralesional injection twice weekly for
8wks

51. Adverse effects of interferon alpha
• Injection site reactions
• Flu-like syndrome
• Thrombocytopenia
• Granulocytopenia
• Depression & suicidal behaviour
• neuropsychiatric symptoms

52. Interferon beta1a & 1b
• Glycoprotein 166 aa
• Recombinant DNA technology
• Treatment of multiple sclerosis
• IFN beta act as Antagonist of endogenous
IL4, IFN gamma
• Trial for intermediate uveitis & macular
edema

53. Interferon beta1a & 1b
• FDA approval - 1996 for Relapsing
Remitting MS
• Clinical trial showed
• slowed MS progression
• slowing or preventing the development of
MS-related brain atropy.

54. Interferon gamma
• Activate phagocytes
• Enhances oxidative metabolism of macrophages
• Enhances ADCC & NKcell activity
• Induces production of MHC1 & MHC2 molecules

55. Interferon gamma
Increases - antimicrobial & tumoricidal
activity of
monocytes
macrophages
neutrophils
NK cells

56. IFN gamma 1b
• Genetically engineered recombinant
• 140 aa
• Used in treatment of malignant osteopetrosis
&
chronic granulomatous disease

57. IFN gamma 1b
• Recent study - treatment option for
patients with refractory multi drug resistant
pulmonary tuberculosis
• Play role - immunity to TB by enhancing
hosts own immune response
• Aerosol form form / SC route

58. Interleukin -2 (aldesleukin)
• Known as Tcell growth factor
• Potent immunomodulator
• Genetically engineered
• treatment of metastatic RCC, metastatic
melanoma

59. Interleukin -2 ( aldesleukin)
• IL2 enhances lymphocyte mitogenesis
• clonal expansion of killer & NK cells -
attack tumor cell .
• Enhances production of TNF,IL 1,IFN
gamma

60. Interleukin -2 ( aldesleukin)
Adverse effect
• capillary leak syndrome- low BP ,
• Decreased renal& lung function
• Change in mental status
• MI , seizure ,coma
• Studies in progress to be used with anti HIV
therapy

61. Generic name Adult dose Route of adm Half life
IFN alpha 2a 3MU 3times
weekly for 12
months
Sc/im 2-5hrs
IFN alpha 2b 5MU od or 10 MU
3times weekly for
16 weeks
Sc/iv/im 2-5 hrs
IFN alpha con 1 9micrograms 3
times weekly for 24
weeks
sc 6-10 hrs
IFN alpha n3 250,000IU per wart
twice weekly for 8
weeks
Intralesional 3-8hrs

62. Peg IFN alpha 2a 180 microgm once
weekly
sc 80 hrs
Anakinra 1-2mg/kg Sc 4-6hrs
Aldesleukin 60,000 iu 8hrly 15
min infusion - `14
doses
Iv 85 min

63. Anticytokines as
therapeutic agents

64. TNF
• primarily involved in the regulation of cell
proliferation and apoptosis
• produced - macrophages, monocytes,
lymphocytes, fibroblasts

65. TNF α
•Also called cachectin
•Produced by macrophages and
Mast cells
•TNF alpha is a pleiotropic inflammatory
cytokine
•Target tissue: tumour cells and
Inflammatory cells
•Has cytotoxic effects
•Induce cytokine secretion
•Responsible for extensive weight loss
(cachexia) associated with chronic
inflammation.
TNF β
•Also called lymphotoxin
•produced by TH1 and Tc cells
•Target tissue: tumour cells and
macrophage and neutrophils
•Has cytotoxic activity and other effects
similar to TNF α
• Enhances the phagocytic activity.

68. TNF alpha
• TNF-α mediates inflammatory processes
in rheumatoid arthritis (RA)by
• immune cell activation,
• proliferation
• apoptosis
• regulation of leukocyte movement.

69. TNF alpha inhibitors
• act by binding to TNF before it binds to
receptors and prevent initiation of
apoptosis or the inflammatory response.
• infliximab
• adalimumab
• etanercept

70. infliximab Etanercept Adalimumab
Structure Chimeric (25%
mouse/75% human)
IgG1 monoclonal
antibody
Recombinant
fusion protein
made of
TNF receptor and
constant Fc
portion of human
IgG1
Fully humanized
anti-TNF
monoclonal IgG1
antibody
Binds to only TNF-α Binds 2 molecules
of TNF-α
as well as TNF-β
Binds only TNF-α
Dose 3–10 mg/kg weekly at
weeks 0, 2, 6;
then every 2 months
25 mg twice a
week
40 mg once in 2
weeks

71. INFLIXIMAB ETANERCEPT ADALIMUMAB
Route of
administrat
ion
IV SC SC
Half-life 9–12 days 4–5 days 10–20 days
Approved
indications
Rheumatoid
arthritis
(methotrexate),
Crohn's disease,
ankylosing
spondylitis
RA (alone
methotrexate),
ankylosing
spondylitis,
juvenile
rheumatoid
arthritis,
psoriatic
arthritis
Moderate to
severe RA
(alone or
methotrexate),
psoriatic
arthritis,
ankylosing
spondylitis,
Crohn's
disease

72. INFLIXIMAB ETANERCEPT ADALIMUMAB
Adverse
effects
Acute infusion
reactions—fever,
chills,
pruritis, chest
pain, dyspnoea,
flushing,
urticaria,
hypersensitivity
reactions,
serious
infections, TB
and sepsis
Less allergic as
compared to
infliximab,
mild injection
site reactions,
serious
infections, TB
and sepsis
Injection site
reactions,
upper
respiratory
infections
(sinus
infections),
headache,
rash, nausea,
serious
infections, TB
and sepsis

73. TNF alpha inhibitors
Contraindication
• previously untreated tuberculosis
• recurrent chest infections
• bronchiectasis
• multiple sclerosis
• demyelinating illness.

74. TNF alpha inhibitors
The scope of therapy is now evolving to
target diseases like
• diabetes,
• psoriasis
• Asthma
• malignancy
• stroke

75. IL 1
• IL1 produced in response to inflammatory
stimuli - inflammation & immune reaction
Increased IL 1 produced in
• Autoimmune diseases
• infections
• solid tumors
• leukemia
• Alzheimer's disease

76. IL 1
• Trauma
• hemodialysis
• ischemic myocardial infarction
• Asthma
• graft versus host disease

77. IL 1
IL1 in pathogenesis of RA
• cartilage degradation by rapid loss of
proteoglycans
• stimulation of bone resorption.

78. IL 1 receptor antagonist -Anakinra
• It is the recombinant non-glycosylated form
of human IL-1 ra
• blocks the biologic activity of IL-1 by
binding to the interleukin-1 type I receptor

79. IL 1 receptor antagonist Anakinra
• Indication -treatment RA unresponsive to one
or more DMARDs.
• Contraindicatioin – active infection
• A/R – injection site reaction
• Monitor for – reversible
thrombocytopenia,neutropenia

80. IL 1 receptor antagonist Anakinra
Under research for –
• cerebral ischemia,
• severe sepsis,
• acute stroke,
• diabetes,
• asthma
• inflammatory bowel disease

81. IL2 receptor antagonist
• Interleukin-2 receptor (IL-2R) is a
heterotrimeric protein composed of α, β and
γ chains
• Receptor expressed - immune cells,
antigen-activated T-cells, B-cells –
proliferation

82. IL2 receptor antagonist
over-expression of IL-2R-α
• autoimmune diseases,
• allograft rejection
• adult T-cell leukemia (ATL),
• mycosis fungoides,

83. IL2 receptor antagonist
• peripheral T-cell lymphoma
• hairy cell leukemia,
• Hodgkins disease
• B-cell neoplasms

84. IL2 receptor antagonist
• monoclonal antibody - basiliximab and
daclizumab
• Indication - Prophylaxis of acute organ
rejection in renal transplant patients

85. basiliximab Daclizumab
antibody Chimeric humanised
Indication Prophylaxis of acute
organ rejection in renal
transplant patients
Prophylaxis of acute
organ rejection in
renal transplant
patients
Halflife 7 days 20 days
Adverse effect Nausea, constipation,
UTI, upper respiratory
tract infection
Nausea,
constipation,
headache

86. Filgrastim (GM-CSF)
• Cancer patients receiving myelosuppresive
chemotherapy
• AML patients receiving induction or
consolidation chemotherapy
• cancer patients receiving
BM transplant,
peripheral blood stem cell transplantation
• severe chronic neutropenia

87. Filgrastim
• Half life – 3-5hrs
• A/E -Skeletal muscle pain,
• bone pain,
• alopecia,
• fever, rash,
• injection site reaction

88. Peg filgrastim (pegylated
GM-CSF)
• Indication – same as filgastrim
• Half life – 45 – 60hrs
• A/E
• Bone pain,
• mylagia, alopecia,
• fever, rash, edema

89. Sargramostim (G-CSF)
Indication
• Following induction chemotherapy- AML
• to accelerate recovery after BM
transplantation

90. Sargramostim (G-CSF)
• Half life – 2-5hrs
• A/E - Bone pain, malaise, fever, diarrhea,
edema, rash

91. Epoietin alpha
Indication
• Anemia - chronic renal failure,
• HIV infected patients,
• chemotherapy,
• primary bone marrow disorders

92. Epoietin alpha
• Half life - 8hrs
• A/E -Hypertension, thrombotic
complication, allergic reactions

93. Darbepoietin alpha
• Indication – same as epoietin
• Half life – 25 hrs
• A/E Mild transient injection site pain
,Hypertension, thrombotic complication,
allergic reactions

94. Oprelvekin
• Recombinant IL 11a /thrombopoietic growth
factor
• stimulates the proliferation of hematopoietic
stem cells and megakaryocyte
• Indication- Chemotherapy induced
thrombocytopenia
• Contraindication – hypersensitivity
decompensated heart failure,renal impairment

95. Oprelvekin
• Dose -50 µg/kg subcutaneously once a day
– abdomen, thigh or hip
• renal impairment- 25 µg/kg daily.
• A/E –allergy , edema ,hypotension ,fever
,papillodema , ventricular arrythmia, renal
failure

96. Ustekinumab
• Human monoclonal antibody directed against
IL12 & IL 23
• Treatment of severe plaque psoriasis
• Phase 2 trial – MS ,Sarcoidosis
• subcutaneous 45 or 90 mg

97. Ustekinumab
• A/E - serious allergic reaction,
• upper respiratory infection,
• headache, and tiredness]
• increased risk of infection- tuberculosis
• increased risk of certain types of cancer

98. Golimumab
• TNF alpha inhibitor
• Once monthly SC
• RA ,Psoriatic arthritis ,ankylosing
spondylitis
• Pending approval for ulcerative colitis
• a/e – injection site reaction
develop infection

99. Canakinumab
• Recombinant anti IL 1 beta monoclonal
antibody
• Indication
systemic juvenile idiopathic arthritis
cryopyrin associated periodic syndrome
• Give subcutaneously
• Dose 2mg/kg , 4mg/kg

100. Rilonacept
• IL 1 inhibitor
• Treatment of gout
• Familial cold auto inflammatory
syndrome
• cryopyrin associated periodic
syndrome

101. Certolizumab pegol
• Monoclonal antibody to TNF
alpha
• Treatment of crohns disease ,RA

102. Tocilizumab
• Humanised IL6 receptor monoclonal
antibody
• rheumatoid arthritis ,systemic juvenile
idiopathic arthritis

103. Mepolizumab
• anti interleukin-5 monoclonal antibody
• used to treat asthma
• reduced - asthma exacerbations, and
reduced the need for corticosteroids

104. Fentolizumab
• Humanized anti- IFN gamma antibody
• Treatment of crohns disease

105. Denileukin Diftitox
• Immunotoxin
• IL2 + Diptheria toxin
• Bind to IL2 receptors - introduce diptheria
toxin in to cells ,killing the cells
• leukemia, lymphoma , cutaneous T cell
lymphoma

106. cytokine therapies in
clinical trial

107. Cytokine Generic name property Phase Potential TU
IL 7 NCT00062049 Dev of pre B
& pre T cell
Phase II HIV induced
lymphopenia
,solid tumours
IL-10 NCT00001761 Inhibit
proinflammator
y
Phase I & II Psoriasis ,scar
reduction ,WG
IL 12 NCT00016289 NK cell
activation
PhaseI,& II Ovarian cancer
,metastatic
melanoma
,NHL

108. CYTOKINE NAME PROPERTY TRIAL POTENTAIL
TU
IL 15 NCTOOOO1761 stimulates
T-cells and NK
cells
Phase I /II Metastatic
melanoma, renal
cell
carcinoma
IL-8 ABX-IL-8 Fully human IL-8
monoclonal
antibody
Phase II Rheumatoid
arthritis,
psoriasis, COPD
IL-9 IL-9mAb Anti-interleukin-
9 monoclonal
antibody
Phase I Asthma
IL-10 Tenovil Recombinant IL-
10 protein
Phase II Crohn's disease

109. CYT NAME PROP TRIAL PTU
TRAP NCT00001145 Induction of
Tcell, Bcell
Phase I
&II
X linked hyper IgM
Syndrome,met
melanoma
IL-12 ABT-874 Humanized
anti-IL 12
monoclonal
antibody
Phase II Moderate to severe
plaque psoriasis,
Crohn's disease,
multiple sclerosis
IL-15 AMG-714 Humanized
anti-IL-15
monoclonal
antibody
Phase II Rheumatoid
arthritis
IL-18 Recombinant
IL-18 binding
protein
Phase II RA, psoriasis

110. CYT NAME PROP TRIAL PTU
IL-21 NCT00336986 Recombinant
human
interleukin-21
Phase II Metastatic
melanoma,renal
cancer,ovarian ca
TRAIL NC00873756 Apoptosis of
tumour cells &
activated T
cells
Phase I Metastatic
colorectal ca,NHL
IL1 AMG 108 Blocking mAB Phase II Osteoarthritis

111. Cytokine Generic name drug Potential TU Phase
IL 4/13 IL 4/ 13 trap recombinant HIV
Asthma
Phase I
IL 5 SCH -55700 antI IL 5 Asthma allergy Phase II
IL 6 MRA antiIL6
receptor
Crohn s disease
SLE
RA
Myeloma
Phase II
Phase I
Phase III
Phase I/II
IL 8 656933 IL 8 antagonist COPD Phase I

112. Future perspectives

113. • will be successful when disease causing
mechanisms are “simple” and cytokine
driven.
• with pleiotropic and redundant nature of
cytokines , it is hard to acheive in complex
disease state

114. • Combination therapies hold great potential
for the future.
• simultaneous inactivation/activation of
multiple cytokines holds good in treating
certain condition

115. • engineered cytokines - undergoing pre-
clinical and clinical evaluation
• likely to constitute the next generation of
cytokine therapeutics
• increased efficacy and reduced incidence of
adverse effects.