2. Who Am I
2nd
Year Doctor of Pharmacy Candidate
Florida Agricultural and Mechanical University
Tallahassee, FL
3. LIFE GOALS
Clinical Trials Pharmacist
Pharmacogenomics- studies the relationship
between a drugs pharmacologic response to a
patients genetic makeup
Assist with Drug Safety, Efficacy, Reduction and
Resolution of Adverse Drug Reactions
4. Kristin + Affinity Team
Perfect Fit!
Laila and Wayne focus on the discovery and
small scale synthesis of Ligands attached to
DNA, and measuring their binding affinities.
Bobby synthesizes the specific Ligands on a
large scale to confirm binding and prepare
protein affinity resins.
5. Kristin + Affinity Team cont.
affinity – natural liking or relationship
Critical Thinking
Problem Solving
6. Organic Synthesis
What kinds of functional groups
are tolerated in the synthesis?
What is the range of different
structures that can be used?
7. Color coding process
4 pieces, represent structures added at the same step in the sequence
Reagents for each step differ
Structure of each compound was confirmed by liquid
chromatography/mass spectrometry (LC/MS)
Purity of compounds is assessed by UV/Vis HPLC and
LC/MS
Mass spectrum is compared against theoretical spectrum to
identify products
NMR is the preferred method for measuring purity, but we
were dealing with insufficient quantities for NMR analysis
Scope of ligand synthesis
8. Ligands that Bind to
Bovine Carbonic Anhydrase (BCA)
Sulfonamides are
known to bind
to BCA
20. Alkene Metathesis
New chemistry for making the library
Library consists of macrocycles synthesized by ring-closing
metathesis (RCM)
8-membered rings through 13-member rings are strained (9 has the
highest, 13 kcal/mol)
14-membered rings and greater have little to no strain (e.g. 15-
member ring has 1.5 kcal/mol strain)
Alkene Metathesis RXN
Ring-closing Metathesis (RCM) RXN
Catalyst
Catalyst
24. Z:Bobby160715160715_HN_RVO_KC_11 07/15/16 10:25:28
RT: 0.00 - 10.01
0 1 2 3 4 5 6 7 8 9 10
Time (min)
0
20
40
60
80
100
RelativeAbundance
RT:7.00
MA:31329293598
BP:282.21
RT:6.29
MA:22127653815
BP:254.17
7.58
282.28
8.13
647.50
NL:
1.61E10
TIC F: FTMS + p
ESI Full ms
[120.00-1800.00]
MS
160715_HN_RVO
_KC_11
200 250 300 350
m/z
0
20
40
60
80
100
0
20
40
60
80
100
RelativeAbundance
254.1747
z=1
226.1799
z=1
310.2371
z=1
335.2786
z=1
284.1852
z=1
196.1694
z=1
362.2295
z=?
310.2377
NL:
1.71E9
160715_HN_RVO_K
C_11#647 RT: 6.29
AV: 1 T: FTMS + p
ESI Full ms
[120.00-1800.00]
NL:
8.12E5
C18 H32 NO3:
C18 H32 N1 O3
pa Chrg 1
Z:Bobby160715160715_HN_RVO_KC_11 07/15/16 10:25:28
RT: 0.00 - 10.01
0 1 2 3 4 5 6 7 8 9 10
Time (min)
0
20
40
60
80
100
RT:7.00
MA:31329293598
BP:282.21
RT:6.29
MA:22127653815
BP:254.17
7.58
282.28
8.13
647.50
NL:
1.61E10
TIC F: FTMS + p
ESI Full ms
[120.00-1800.00]
MS
160715_HN_RVO
_KC_11
100
254.1747
z=1
NL:
1.71E9
Alkene Metathesis
Hoveyda Grubbs Second Generation Catalyst
~40% conversion
13-membered ring 5 kcal/mol ring strain decreased the conversion rate
25. SEPARATING ALKENES BY silver nitrate silica
Silver Nitrate silica allows separation of alkenes that are
inseparable by regular silica chromatography
Silver Nitrate non covalently binds to alkenes
Since the starting material contains two alkenes it should
move slower up the plate in comparison to the final product,
which has one alkene.
27. A very friendly environment
Witnessing the companies progression
The Affinity Team was very encouraging
Unique and genuine personalities collaborated on
several levels
What I Liked Best About Working Here
28. There is a place for everybody at Impossible Foods
Degrees and Work Experiences vary extensively, and
Impossible is home to all.
What I found most interesting
7 structures were chosen from a list that met the criteria of
Acylation, Amination, Addition
Add desired MW
Aromatic Ring- decreased the likelihood of going all the way to the product because of the other resonance structures
full ligand missing possibly because of solid support resin decomposition… there was immediate discoloration when the resin was added; with all of the other compounds this was not observed
Full conversion
Steps A, B, and C worked but step D didn’t
Carboxylic acid containing amines did notdissolve well with THF; possibly a solubility problem not reactivity
