Sidechain Tyrosine Resin Loading

Loading Fmoc O Allyl Tyrosine

applicable for sequences bound for on resin cyclization

Overall Reaction

This method describes the loading of tyrosine through the sidechain for the purposes of on-resin cyclization. This is chemistry allows for on resin cyclization and global methylation. It is however not stable to the reducing conditions used for either NBS or TFA mediated in sequence selective methylation. In contrast to resin bound for solution phase cyclization this resin is not loaded to maximum capacity, instead to prevent dimerization during cyclization it is loaded no higher than 0.45 mMol/g. This method was adopted from Arthur Rand and Rushia Turner.[1]

This page assumes the loading of 5 grams of 2-Chlorotrityl resin

Step 1

Tyrosine Preparation
  1. Take an oven dried 250 mL round bottom flask and cool it under a stream of nitrogen
  2. Add a 2.25 mMol (0.99 g) of Fmoc O Allyl Tyrosine to the flask (previously prepared or purchased)
  3. Rubberband a kimwipe on top and vaccuum desicate overnight.

Step 2

  1. Acquire ~100 mL of dry DCM from the solvent purifcation system
  2. Add ~75 mL of DCM (via syringe or cannula) to the flask charged with Tyrosine
  3. Add 4 eq (1.17 mL) of sieve dried DIPEA
  4. Dissolve as best you can
  5. Add 5 g of unloaded 2 Chlorotrityl resin
  6. Add a head of nitrogen (blowing away the white smoke that forms)
  7. Cap with a septa and a nitrogen balloon.
  8. Shake overnight on the orbital shaker
  • PRO-TIP slowly rolling the flask will collect stray resin that collections above the solvent line

Example Reaction Table
Name g MW mmol eq d mL
2-Cl-Trytl resin 5 n/a n/a n/a n/a n/a
Fmoc-OAllyl-Tyrosine 1 443.41 2.25 1 n/a n/a
DIPEA 1.2 129 9 5 0.74 1.6

Step 3

Collecting Resin
  1. Collect the resin into a solid phase synthesis vessel using DCM and a funnel
  2. Wash 3x with DCM
Prepare Capping Solution
  1. Make a 17:2:1 solution of DCM:MeOH:DIPEA
  2. For this scale 51 mL of DCM, 6 mL of MeOH, and 3 mL of DIPEA works great
  1. Add 20 mL of capping solution, mix briefly, drain
  2. Add 20 mL and shake for 20 minutes
  3. Add the final 20 mL and shake for another 20 minutes
  • PROTIP: Make sure to release the pressure from the synthesis vial before shaking or the cap will eject
  1. Wash 3x DMF
  2. Wash 3x DCM
  3. Put a rubberbanded kimwipe on top and pull air over the resin overnight for future loading determination

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