pe
pep-10415 v1 CC-BY-SA-4.0

Pain-receptor research peptide (CHEMBL1161419)

A tiny lab-made peptide used to study how opioid receptors, the same ones morphine acts on, recognize pain-relieving molecules; a research tool only, not a medicine.

statusbioassayed targetOPRD1 length3 aa refs1
status 5 / 5
prediction metrics boltz-2 1.0
ipTM0.917
pTM0.839
avg pLDDT81.4
ranking score0.834
STRUCTURE · PEP-10415 × OPRD1
ranking0.834
target interface 4.5Å peptide drag rotate · ctrl+scroll zoom · right-click pan
boltz-2 1.0 · mmCIF ↓ download
sequence3 aa
13
FFF
overview readme

What this is

CHEMBL1161419 is a synthetic four-residue research peptide designed to probe how the mu-opioid receptor — the main target of morphine and related analgesics — recognises its ligands. Built as a modified version of endomorphin-2, a naturally occurring brain opioid peptide, it was synthesised at Harvard as part of a structure–activity study examining what structural features are needed for high-affinity mu-receptor binding. The compound binds the mu-opioid receptor (OPRM1) with sub-nanomolar potency while showing essentially no affinity for the delta-opioid receptor (OPRD1), despite being catalogued in ChEMBL under OPRD1 because a delta-receptor assay was also conducted. It is not approved for any use and exists only as a biochemical tool. The stored raw sequence (FFF) represents only the three phenylalanine residues; the full compound is a tetrapeptide with D-proline at position 2 and a C-terminal amide, neither of which appears in the stored sequence field.

History

Endomorphin-2 (H-Tyr-Pro-Phe-Phe-NH₂) is an endogenous tetrapeptide first described in 1997 that shows unusually high selectivity for the mu-opioid receptor. Harrison and colleagues at Harvard (Verdine laboratory) used endomorphin-2 as the scaffold for an exhaustively stereodiversified ligand library to explore the chemical space around its core backbone. In a 2003 publication in the Journal of Medicinal Chemistry, they reported a series of analogues — including compounds with 2,6-dimethyltyrosine at position 1 and varied stereochemistry at other positions — that retained or exceeded endomorphin-2's potency at OPRM1 (Harrison and colleagues, J Med Chem 2003). CHEMBL1161419, with phenylalanine substituted at the N-terminal position in place of the canonical tyrosine and D-proline at position 2, was among the compounds characterised in that programme to establish how each structural departure from the natural sequence affects receptor selectivity and binding affinity.

What it does

The compound binds the mu-opioid receptor with high affinity and acts as a full agonist: in a [³⁵S]GTPγS functional assay, it stimulates G-protein coupling in OPRM1-expressing cell membranes with an EC50 of 31 nM and 100% maximal efficacy relative to a reference agonist (Harrison and colleagues, J Med Chem 2003). It shows negligible affinity for the delta-opioid receptor and low affinity for the kappa-opioid receptor at the concentrations studied. Its primary scientific value lies in the SAR insight it provides: by replacing the N-terminal tyrosine of endomorphin-2 with phenylalanine — thereby removing the phenolic hydroxyl that was traditionally considered essential for opioid activity — researchers could quantify the hydroxyl's contribution to mu selectivity and binding potency.

Evidence

  • Human: No human studies. This is a research ligand only.
  • Animal: No in vivo animal studies identified for this specific compound.
  • In vitro: Radioligand displacement assays in transfected cell lines: OPRM1 Ki = 0.69 nM (displacement of [³H]DAMGO from human OPRM1-expressing CHO cells); OPRD1 Ki = 25,000 nM (displacement of [³H]diprenorphine from human OPRD1-expressing HEK293 cells); OPRK1 Ki = 10,400 nM (guinea pig cerebellar preparation). The μ/δ selectivity ratio is approximately 36,000-fold in favour of OPRM1. Full-agonist functional activity confirmed by [³⁵S]GTPγS incorporation. All data from Harrison and colleagues (J Med Chem 2003).

Known effects

  • OPRM1 binding — High potency in vitro (Ki 0.69 nM); full agonist by [³⁵S]GTPγS assay (EC50 31 nM). Mechanistic only — no in vivo efficacy data for this compound.
  • OPRD1 binding — Essentially inactive (Ki 25,000 nM). The ChEMBL record indexes this compound under OPRD1 because the delta-receptor displacement assay was run, but the measured affinity is negligible.
  • OPRK1 binding — Weak (Ki 10,400 nM).

Mechanism

CHEMBL1161419 belongs to the endomorphin-2 analogue class. Endomorphin-2 (Tyr-Pro-Phe-Phe-NH₂) achieves its exceptional mu-selectivity through several structural features: the N-terminal tyrosine phenolic hydroxyl makes key contacts in the OPRM1 orthosteric binding pocket; the L-Pro² residue constrains backbone conformation; and the C-terminal amide protects against carboxypeptidase degradation. In CHEMBL1161419, Tyr¹ is replaced by Phe (removing the hydroxyl) and L-Pro² is replaced by D-Pro², inverting stereochemistry at that position. Despite removal of the phenolic hydroxyl — a modification that in other endomorphin-2 analogue series substantially reduces mu-receptor affinity — this compound retains sub-nanomolar potency at OPRM1. The D-Pro substitution additionally improves resistance to dipeptidyl peptidase IV, the principal enzyme that cleaves at the Pro² position of endomorphins. Upon mu receptor engagement, the compound couples to Gi/o proteins, inhibiting adenylyl cyclase, suppressing N- and P/Q-type calcium channel conductance, and activating inwardly rectifying potassium channels — the same downstream pathway as morphine and endogenous enkephalins.

Regulatory status

  • US: Not approved. Research reagent only.
  • EU: Not approved. Research reagent only.
  • No registered trials on ClinicalTrials.gov for this compound.

Related peptides

  • Endomorphin-2 analogue with cyclic scaffold — pep-10425: another research ligand built on the endomorphin-2 Tyr-Pro-Phe-Phe-NH₂ core, here constrained into a cyclic pseudo-tetrapeptide; studied in the same opioid receptor SAR context.
  • CTAP-type mu-opioid research ligand — pep-10434: a five-residue OPRM1 binder from the somatostatin-template CTAP scaffold series; a structurally distinct approach to mu-receptor pharmacology studied in parallel SAR programmes.
Hypotheses5 directions▾ collapse

Research directions for this peptide, selected from the current sources — hypotheses you can explore and model. None of it is proven yet; tap any one to see the full thinking.

openupdated 2026-06-05

Does flipping one amino acid from its unusual mirror-image form back to normal collapse selectivity for the pain receptor?

If true, drug designers chasing non-addictive painkillers could focus on mimicking a geometric kink rather than endlessly varying bulky side groups, potentially speeding up the discovery of safer analgesics for the millions living with chronic pain.

The hypothesis
The mu-selectivity of this endomorphin-2 analogue over delta-opioid receptor is primarily determined by the D-Pro2 stereochemical constraint rather than the tri-phenylalanine aromatic cluster, such that replacing D-Pro2 with L-Pro2 would collapse selectivity even while retaining high aromatic bulk at positions 3-4.
Why it’s plausible
The stored sequence FFF captures only the three phenylalanine residues; the full compound is H-Tyr-D-Pro-Phe-Phe-NH2 with sub-nanomolar OPRM1 affinity and negligible OPRD1 affinity. OPRM1 and OPRD1 share high orthosteric pocket homology, so the aromatic residues alone are unlikely to encode selectivity. D-Pro2 introduces a rigid type-II' beta-turn geometry that positions the Tyr1 pharmacophore and the C-terminal Phe-Phe dipeptide in an OPRM1-complementary orientation. Endomorphin-2 SAR literature documents loss of mu-selectivity upon Pro stereochemical inversion, consistent with a conformational rather than contact-residue basis for selectivity.
Why it matters
Identifying the backbone constraint as the selectivity determinant rather than the aromatic side chains reframes the design logic for non-addictive analgesics: rigid turn mimetics, not aromatic substitution, would be the productive engineering axis.
Plausibility.78
Novelty.55
Impact.70
Basis · grounding1 paper · 2 computed/notes
[1]
paper
Verdine lab SAR series using endomorphin-2 scaffold with stereodiversified analogues; selective OPRM1 binders identified among the library
doi: 10.1021/jm025608s
[2]
noteFull compound is Tyr-D-Pro-Phe-Phe-NH2; sub-nanomolar OPRM1 potency with essentially no OPRD1 affinity despite OPRD1 assay being run
[3]
sequenceStored sequence FFF omits the D-Pro2 and Tyr1 residues, confirming the selectivity-critical residues are outside the minimal stored motif
openupdated 2026-06-05

Does the amide group at the end of this peptide actively trigger receptor activation, or just stabilise the molecule?

Understanding which chemical features flip a receptor from active to inactive could guide the rational design of opioid antagonists for overdose reversal or opioid-use disorder treatment, helping clinicians with more targeted options.

The hypothesis
The C-terminal amide of this compound, absent from the stored FFF sequence, is required not just for metabolic stability but for a direct hydrogen-bond contact with the OPRM1 Gln124 or equivalent residue that stabilises the active-state receptor conformation, such that removing the amide and restoring the free carboxylate would convert the compound from a potent agonist to a partial agonist or neutral antagonist.
Why it’s plausible
Endomorphin-2 and its analogues carry a C-terminal amide (NH2), which is common among endogenous neuropeptides and bioactive opioid peptides. In OPRM1 crystal structures and MD simulations, the carboxylate/amide terminus of peptide ligands often contacts polar residues in the binding pocket that are coupled to the toggle switch (Trp293 rotamer) associated with receptor activation. The distinction between free acid and amide at the C-terminus changes both charge state and hydrogen-bond donor/acceptor character at physiological pH, which could shift the receptor's conformational equilibrium. The stored FFF sequence lacks the amide notation, meaning a naive synthesis following the stored sequence would produce a free-acid compound with potentially different efficacy.
Why it matters
Establishing the C-terminal amide as an efficacy switch rather than merely a stability feature would mean that sequence-database-derived peptide synthesis without full chemical annotation would produce pharmacologically distinct compounds, a practical warning for peptide library work.
Plausibility.70
Novelty.50
Impact.65
Basis · grounding1 paper · 2 computed/notes
[1]
noteFull compound is Tyr-D-Pro-Phe-Phe-NH2; C-terminal amide is specified as part of the active structure; stored FFF sequence omits this feature
[2]
noteCompound described as a biochemical tool for studying OPRM1 recognition; precise chemical features including amide are essential to the binding pharmacology
[3]
paper
GTP-gamma-35S G-protein coupling assay measures functional agonism, providing a readout sensitive to partial vs full activation states that would differ between amide and free-acid forms
doi: 10.1021/jm025608s
openupdated 2026-06-05

Would adding small methyl groups to the ring structures of the phenylalanine amino acids push binding to the pain receptor from nanomolar down to picomolar strength?

If true, an ultra-potent version of this scaffold could work at much lower doses, potentially reducing side-effect burden for patients requiring strong analgesics and giving chemists a cleaner tool to study opioid receptor biology.

The hypothesis
Replacing the three natural L-Phe residues with 2,6-dimethyl-L-phenylalanine (Dmp) at positions 3 and/or 4 of the full tetrapeptide will increase OPRM1 affinity beyond the sub-nanomolar baseline by filling a hydrophobic sub-pocket adjacent to the canonical aromatic cage, because the Verdine lab series already demonstrated that 2,6-dimethyltyrosine at position 1 enhances binding, implying that aromatic ring methylation is tolerated and potentially beneficial throughout the scaffold.
Why it’s plausible
The Harvard 2003 SAR series used 2,6-dimethyltyrosine (Dmt) at position 1 to enhance affinity; this substitution adds steric bulk ortho to the hydroxyl, improving van der Waals contacts with the OPRM1 pocket. If the same principle applies to the C-terminal Phe residues, which contribute the FFF triad captured in the stored sequence, then Dmp substitution at Phe3 or Phe4 could push affinity into the picomolar range. The high ipTM from boltz-2 (0.917) for the FFF fragment suggests the pocket can accommodate the aromatic triad, and ortho-methylation would add contacts without clashing if the pocket geometry is favorable.
Why it matters
Picomolar OPRM1 agonists with a rigid turn scaffold and metabolic stability from D-amino acids and methylated aromatics represent an unexplored region of analgesic chemical space with potential for ultra-low dose activity.
Plausibility.55
Novelty.60
Impact.60
Basis · grounding1 paper · 2 computed/notes
[1]
paper
Verdine lab series includes 2,6-dimethyltyrosine at position 1 of endomorphin-2 analogues to enhance OPRM1 affinity; systematic aromatic modification strategy documented
doi: 10.1021/jm025608s
[2]
noteFull compound is a modified endomorphin-2 with enhanced mu-receptor affinity; scaffold is explicitly SAR-optimised
[3]
structureboltz-2 ipTM=0.917 for FFF fragment indicates the aromatic triad fits the receptor pocket, suggesting room for productive steric augmentation
openupdated 2026-06-05

Could this compound accidentally bind the anti-opioid receptor NPFFR2, which controls how quickly pain drugs lose their effect?

If this peptide hits both receptors, it could reduce the tolerance build-up that forces chronic pain patients onto ever-higher opioid doses, potentially offering a new strategy to maintain long-term pain relief without escalating drug exposure.

The hypothesis
The Tyr-D-Pro-Phe-Phe-NH2 scaffold, optimised for OPRM1, may bind the neuropeptide FF receptor 2 (NPFFR2) with measurable affinity because NPFFR2 recognises short aromatic C-terminal amide peptides (canonical motif PQRFamide) and shares a class-A GPCR aromatic binding pocket architecture with opioid receptors, suggesting that this compound could modulate opioid-induced hyperalgesia through an NPFFR2 mechanism independent of its direct OPRM1 agonism.
Why it’s plausible
NPFFR2 is an anti-opioid peptide receptor that binds RF-amide peptides; its endogenous ligands are short (4-8 residues) with C-terminal Phe-amide or Arg-Phe-amide motifs. The compound's Phe-Phe-NH2 C-terminus is structurally similar to this recognition element. Opioid-induced hyperalgesia is partly mediated through NPFF receptor upregulation, and a compound that simultaneously activates OPRM1 and modulates NPFFR2 could have a self-limiting analgesic profile. This cross-reactivity would be unexpected from the compound's design intent and would only surface in a broad receptor panel screen.
Why it matters
If confirmed, this scaffold would be the first documented dual OPRM1 agonist/NPFFR2 ligand derived from endomorphin-2, providing a molecular probe for the opioid-anti-opioid balance and a potential lead for reducing opioid tolerance.
Plausibility.45
Novelty.70
Impact.60
Basis · grounding3 computed/notes
[1]
sequencePhe-Phe-NH2 C-terminus of the full compound overlaps with the Phe-amide recognition motif of NPFFR2 endogenous ligands
[2]
noteCompound is a synthetic endomorphin-2 analogue; endomorphin-2 itself has not been systematically screened against the RF-amide receptor family
[3]
structureboltz-2 ipTM=0.917 for FFF in complex with opioid receptor confirms the aromatic triad is a productive binding unit; the same motif is present in NPFF family ligands
openupdated 2026-06-05

Is the gap between this compound's activity at the two opioid receptors large enough to separate pain relief from delta-receptor side effects?

If the potency gap is as large as the data suggest, this scaffold could inform analgesic designs that deliver pain relief with fewer delta-opioid complications, potentially benefiting patients who need long-term opioid therapy.

The hypothesis
The ChEMBL annotation of this compound under OPRD1 is a cataloguing artefact from a counter-selectivity assay, and the compound has measurable but low-affinity OPRD1 partial-agonist activity at micromolar concentrations that is functionally distinct from its sub-nanomolar OPRM1 full-agonism, creating a built-in therapeutic window separating analgesia from delta-mediated side effects.
Why it’s plausible
The readme explicitly states the compound binds OPRM1 with sub-nanomolar potency and shows essentially no affinity for OPRD1, yet it is catalogued under OPRD1 because a delta assay was run. However, 'essentially no affinity' in competitive binding does not exclude low-level functional activity: at 10 uM (the GTP-gamma-S assay concentration noted in the literature snippet from 10.1021/jm025608s), compounds fully saturate receptors and any residual OPRD1 agonism would become detectable. Delta-opioid receptor activity is associated with anxiolytic effects but also seizure risk at high doses; a compound with a 1000-fold or greater OPRM1/OPRD1 potency ratio would have a built-in pharmacological separation that is clinically desirable.
Why it matters
If the potency ratio is quantified, this scaffold becomes a benchmark for mu-selective tool compounds and the D-Pro2 constraint a validated design element for separating analgesic from delta-mediated liabilities.
Plausibility.60
Novelty.35
Impact.50
Basis · grounding1 paper · 2 computed/notes
[1]
paper
GTP-gamma-35S assay run at 10 uM to assess G-protein coupling via MOR; same study tested selectivity across receptor subtypes
doi: 10.1021/jm025608s
[2]
noteCompound annotated under OPRD1 in ChEMBL solely because a delta-assay was conducted; actual OPRD1 affinity described as essentially absent
[3]
noteSub-nanomolar OPRM1 potency established; selectivity vs OPRD1 is the defining pharmacological feature of this analogue series
details expand to inspect
full evidence table1 metrics
metricvaluetool
Ki 25000 nM GPCRDB/ChEMBL
structural qualityopenfold3
metricvaluenote
gpde0.949global PDE — lower = better
disorderNaNfraction disordered
3-letter notation
Phe-Phe-Phe
recipeboltz-2 1.0
parametervalue
modelboltz-2 1.0
weights
hardwarenvidia_nim_api
mlx version
python
random seed
msa strategynone
diffusion samples1
runtime
predicted bymlx@peptide
predicted at2026-04-24
citationbibtex
peptidemodel (2026). Pain-receptor research peptide (CHEMBL1161419) (pep-10415, v1). PeptideModel. https://peptidemodel.com/card/pep-10415
@peptide{pep10415,
  sequence = {FFF},
  target   = {oprd1},
  author   = {peptidemodel},
  year     = {2026},
  status   = {bioassayed}
}
clinical trials 0 trials · checked 2026-05-22
0
no registered clinical trials as of 2026-05-22; we'll re-check periodically
references 1 papers
discussion no comments
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peptidemodel.com CC-BY-SA-4.0 research only · not for human use